凋亡调控蛋白Bcl-2、Bax与非酒精性脂肪性肝病的关系及药物干预研究
|
摘要
目的：非酒精性脂肪性肝病(NAFLD)是以甘油三酯在肝内过度贮积而引起的临床病理综合征,其范畴包括单纯性脂肪肝(NASFL)、非酒精性脂肪性肝炎(NASH)和NASH相关性肝硬化。目前非酒精性脂肪肝的发病机制尚不明确,以Day等提出的“二次打击”学说得到普遍公认。近年来随着对细胞凋亡的研究,不断有证据表明：在NAFLD中肝细胞凋亡可能是单纯非酒精性脂肪肝发展为非酒精性脂肪性肝炎(NASH)“二次打击”理论的关键步骤,也是该过程的重要标志,与炎症程度和纤维化进展相关。在细胞凋亡的两个进化保守的信号转导途径中,Bcl-2家族成员的构成比例是凋亡调控的关键因素,尤其这一家族的两个代表性成员Bcl-2和Bax是Bcl-2基因家族中分别抑制和促进肝细胞凋亡的结构相似、作用相反的一对胞内蛋白质,其比值直接决定肝细胞接受死亡信息后,细胞是走向生存还是死亡的命运。本实验采用高脂胆固醇饮食建立非酒精性脂肪肝大鼠模型,观察大鼠肝组织中凋亡调控蛋白Bcl-2、Bax的活性和基因表达情况,对Bcl-2、Bax在NAFLD发病机理中的作用进行初步探讨,同时观察降脂益肝冲剂对肝组织Bcl-2 Bax基因表达的影响,探讨其可能的分子治疗机制,为临床应用提供理论依据。
方法：1.动物模型的建立及标本的采集：70只雄性Wistar大鼠正常喂养一周后,随机分为正常对照组(N组)：给予正常饲料；模型对照组(M组),用药组(D组)：给予高脂饲料(85%普通饲料,13%的猪油和2%的胆固醇),实验设4、8、12周三个时相点,相对应的组别分别为N1、N2、N3,M1、M2、M3,D组,每组各10只。于实验第8周末,D组开始给予降脂益肝冲剂1ml／100g,以0.8g/ml的浓度溶解灌胃,每日两次,持续4W。同时M组和N组分别给予等量的生活饮用水灌胃。分阶段处死各组大鼠,自腹主动脉采血,迅速取出肝脏,按常规制备血清及肝组织切片,并留取少量肝组织于-70℃低温保存。
2.检测指标：测体重、体长、肝湿重,并计算肝指数(肝湿重／体重×100%)、Lee's指数(体重0.33/体长×1000)等体质指标；全自动生化分析仪测定丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、血脂含量；光镜下评估肝脂肪变性、炎症活动度和纤维化程度；蛋白印迹法(Western Blot)检测肝组织中凋亡调控蛋白Bcl-2、Bax的表达。
结果：1.N组大鼠肝小叶结构完整,肝索围绕中央静脉呈放射状排列,细胞呈多边形,大小一致；各项生化指标均在正常范围；肝组织内可见凋亡调控蛋白Bcl-2、Bax的表达,且二者比值保持平衡。
2.4周末,M1组大鼠出现轻度脂肪变,无炎性细胞浸润,无纤维化；血清转氨酶水平、血脂含量较N1组升高；肝组织凋亡调控蛋白Bcl-2表达下降,Bax表达增多,二者比值下降(P<0.01)。
3.8周末,M2组大鼠肝组织呈单纯性脂肪肝改变,无炎性细胞浸润,纤维化S1期；血清转氨酶水平、血脂含量较N2组明显升高；肝组织凋亡调控蛋白Bcl-2表达较N2组显著下降,Bax表达增多,二者比值下降(P<0.01)。
4.12周末,M3组大鼠肝细胞呈弥漫性脂肪变,伴大量炎性细胞浸润,肝纤维化S2期；血清转氨酶水平、血脂含量较N3组显著升高；肝组织凋亡调控蛋白Bcl-2表达较N3组显著下降,Bax表达增多,二者比值更低(P<0.01)。D组大鼠肝脂变程度、炎症活动度、纤维化较M3组明显减轻；血清学各项指标均有显著改善；(P<0.01或P<0.05)；肝组织Bcl-2表达较M3组明显增多,Bax表达减少,二者比值升高,但仍较N3组低(P均<0.01)。
结论：1.高脂高胆固醇饮食可以成功复制NAFLD大鼠模型,脂质代谢紊乱、炎症损伤以及肝细胞凋亡与NAFLD的发生密切相关。
2.正常大鼠肝脏表达一定量的Bcl-2、Bax蛋白,且二者比值Bcl-2、Bax保持平衡,NAFLD大鼠肝脏Bcl-2蛋白表达下调,Bax蛋白表达增多,Bcl-2、Bax降低,且随造模时间延长Bcl-2、Bax两者比值显著下降。
3.降脂益肝冲剂对NAFLD具有良好的防治效果,其发挥治疗作用可能与上调肝脏Bcl-2蛋白表达,减少Bax蛋白,纠正二者平衡失调有关。
Objective:NAFLD is a clinical syndrome, and has a spectrum ranging from that of fatty liver alone to nonalcoholic steatohepatitis (NASH), which may progress to liver cirrhosis and can progress to liver cell necrosis, fibrosis and cirrhosis of the liver. At present the mechanisms underlying NAFLD are focus on "two hits" theory posed by Day. With the study of apoptosis, more and more researches demonstrate that apoptosis is the key steps when fatty liver alone progress to nonalcoholic steatohepatitis (NASH), and closely related to inflammation、fibrosis. In the two of apoptosis signal transduction pathways, Bcl-2 family members' ratio is the key factor, especially the most representative members Bcl-2 and Bax, which have the simily structure but the opposite function, the ratio is directly determines the fate of liver cells when they accept the death message, the cell towards survival or death. In this study, we established rat NAFLD model by high-fat and high-cholesterol diet to try to explore the following vievpoint:(1)To observe the expression of apoptosis-regulated proteins Bcl-2、Bax in liver of rats in every stage of NAFLD model; (2)To investigate whether Jiangzhi Yigan granules can treat and prevent NAFLD through regulating the expression of apoptosis-regulated proteins Bcl-2、Bax or not. This investigation may provide new clues for the pathogenesis of NAFLD and instruct clinical medication.
Methods:1. Animal Model and Specimen Collection:70 male Wistar rats fed a normal diet after one week, were randomly divided into normal control group (N group):normal diet; the model group (M group), drug treatment group (D group):given high fat diet (85% normal diet, 13% lard and 2% cholesterol). The experimental set up 4,8,12 weeks time point,10 rats for each time point, groups corresponding to the N1, N2, N3, M1, M2, M3, D. Drug treatment group was given Jiangzhi Yigan Chongji (1ml/100g rat body weight, its concentration is 0.8g/ml, twice every day) by intragastric administration separately after the end of 8th week, meanwhile the normal and model group were separately given life drinking water by the same means. All the rats were sacrificed after 4,8,12 weeks. Since the abdominal aortic blood, quickly remove the liver, according to conventional preparation of serum, liver homogenate, and to retain a small amount of liver tissue cryopreservation in-70℃.
2. Detect indicators:measured body weight, body length, wet weight of liver, and calculate the liver index (liver wet weight/body weight×100%), Lee's index (0.33 weight/body length×1000) and other physical indicators. Automatic biochemical analyzer determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood lipids content, light microscope to assess steatosis, inflammatory activity and fibrosis; The levels of Bcl-2 and Bax expression were determined by Western Blot.
Results:1. The liver lobules of normal group rats structural integrity, liver cord around the central venous actinomorphous distribution, cells were polygonal, various biochemical indices were within the normal range; the rates of Bcl-2 and Bax keep the balance.
2. After 4 weeks, the model rats exhibited mild steatosis, no inflammatory cell infiltrationand and no fibrosis; increase of aminotransferase; compared with normal group, the expression of Bax protein was more pronounced, while the expression of Bcl-2 was decreased, Bcl-2/Bax was also decreased significantly (P<0.01).
3. After 8 weeks, control with the N2 group, model group M2 rats, simple fatty liver were observed, no inflammatory cell infiltration and fibrosis S1; serum transaminase levels were significantly higher than normal, TC, TG, ALT, AST; liver Bax expression than the control group increased significantly, while Bcl-2, Bcl-2/Bax decreased (P<0.01).
4. After 12 weeks, severe steatohepatitis with fibrosis S2 of M3 rats; compared with normal group, the level of aminotransferase and blood fat were increased in model group; liver Bax expression than the control group increased significantly, while Bcl-2, Bcl-2/Bax decreased; compared with M3 group, the treated group every biochemical manifestation shew improvement significantly, the expression of Bax in liver was decreased, Bcl-2 and Bcl-2/Bax increased; but still lower than normal group.
Conclusion:1. High-fat and high-cholesterol diet can successfully copy the NAFLD rat model; lipid metabolism disorders, inflammatory injury and apoptosis are closely related to the occurrence of NAFLD.
2. Bcl-2 and Bax can be expressed in normal rat liver, the expression of Bax protein was more pronounced from 4 week and continued to rise until 8 week and 12 week in the model group, compared with the normal group the expression of Bcl-2 was decreased, Bcl-2/Bax in the model group, particularly at 12 week decreased in a time-dependent manner.
3. Jiangzhi Yigan granule has a good anti-therapeutic effects for combat NAFLD. Its molecular mechanism may be partly through up-regulating the expression of Bcl-2 of the live, down-regulating Bax and keeping the balance of Bcl-2 and Bax.
方法：1.动物模型的建立及标本的采集：70只雄性Wistar大鼠正常喂养一周后,随机分为正常对照组(N组)：给予正常饲料；模型对照组(M组),用药组(D组)：给予高脂饲料(85%普通饲料,13%的猪油和2%的胆固醇),实验设4、8、12周三个时相点,相对应的组别分别为N1、N2、N3,M1、M2、M3,D组,每组各10只。于实验第8周末,D组开始给予降脂益肝冲剂1ml／100g,以0.8g/ml的浓度溶解灌胃,每日两次,持续4W。同时M组和N组分别给予等量的生活饮用水灌胃。分阶段处死各组大鼠,自腹主动脉采血,迅速取出肝脏,按常规制备血清及肝组织切片,并留取少量肝组织于-70℃低温保存。
2.检测指标：测体重、体长、肝湿重,并计算肝指数(肝湿重／体重×100%)、Lee's指数(体重0.33/体长×1000)等体质指标；全自动生化分析仪测定丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、血脂含量；光镜下评估肝脂肪变性、炎症活动度和纤维化程度；蛋白印迹法(Western Blot)检测肝组织中凋亡调控蛋白Bcl-2、Bax的表达。
结果：1.N组大鼠肝小叶结构完整,肝索围绕中央静脉呈放射状排列,细胞呈多边形,大小一致；各项生化指标均在正常范围；肝组织内可见凋亡调控蛋白Bcl-2、Bax的表达,且二者比值保持平衡。
2.4周末,M1组大鼠出现轻度脂肪变,无炎性细胞浸润,无纤维化；血清转氨酶水平、血脂含量较N1组升高；肝组织凋亡调控蛋白Bcl-2表达下降,Bax表达增多,二者比值下降(P<0.01)。
3.8周末,M2组大鼠肝组织呈单纯性脂肪肝改变,无炎性细胞浸润,纤维化S1期；血清转氨酶水平、血脂含量较N2组明显升高；肝组织凋亡调控蛋白Bcl-2表达较N2组显著下降,Bax表达增多,二者比值下降(P<0.01)。
4.12周末,M3组大鼠肝细胞呈弥漫性脂肪变,伴大量炎性细胞浸润,肝纤维化S2期；血清转氨酶水平、血脂含量较N3组显著升高；肝组织凋亡调控蛋白Bcl-2表达较N3组显著下降,Bax表达增多,二者比值更低(P<0.01)。D组大鼠肝脂变程度、炎症活动度、纤维化较M3组明显减轻；血清学各项指标均有显著改善；(P<0.01或P<0.05)；肝组织Bcl-2表达较M3组明显增多,Bax表达减少,二者比值升高,但仍较N3组低(P均<0.01)。
结论：1.高脂高胆固醇饮食可以成功复制NAFLD大鼠模型,脂质代谢紊乱、炎症损伤以及肝细胞凋亡与NAFLD的发生密切相关。
2.正常大鼠肝脏表达一定量的Bcl-2、Bax蛋白,且二者比值Bcl-2、Bax保持平衡,NAFLD大鼠肝脏Bcl-2蛋白表达下调,Bax蛋白表达增多,Bcl-2、Bax降低,且随造模时间延长Bcl-2、Bax两者比值显著下降。
3.降脂益肝冲剂对NAFLD具有良好的防治效果,其发挥治疗作用可能与上调肝脏Bcl-2蛋白表达,减少Bax蛋白,纠正二者平衡失调有关。
Objective:NAFLD is a clinical syndrome, and has a spectrum ranging from that of fatty liver alone to nonalcoholic steatohepatitis (NASH), which may progress to liver cirrhosis and can progress to liver cell necrosis, fibrosis and cirrhosis of the liver. At present the mechanisms underlying NAFLD are focus on "two hits" theory posed by Day. With the study of apoptosis, more and more researches demonstrate that apoptosis is the key steps when fatty liver alone progress to nonalcoholic steatohepatitis (NASH), and closely related to inflammation、fibrosis. In the two of apoptosis signal transduction pathways, Bcl-2 family members' ratio is the key factor, especially the most representative members Bcl-2 and Bax, which have the simily structure but the opposite function, the ratio is directly determines the fate of liver cells when they accept the death message, the cell towards survival or death. In this study, we established rat NAFLD model by high-fat and high-cholesterol diet to try to explore the following vievpoint:(1)To observe the expression of apoptosis-regulated proteins Bcl-2、Bax in liver of rats in every stage of NAFLD model; (2)To investigate whether Jiangzhi Yigan granules can treat and prevent NAFLD through regulating the expression of apoptosis-regulated proteins Bcl-2、Bax or not. This investigation may provide new clues for the pathogenesis of NAFLD and instruct clinical medication.
Methods:1. Animal Model and Specimen Collection:70 male Wistar rats fed a normal diet after one week, were randomly divided into normal control group (N group):normal diet; the model group (M group), drug treatment group (D group):given high fat diet (85% normal diet, 13% lard and 2% cholesterol). The experimental set up 4,8,12 weeks time point,10 rats for each time point, groups corresponding to the N1, N2, N3, M1, M2, M3, D. Drug treatment group was given Jiangzhi Yigan Chongji (1ml/100g rat body weight, its concentration is 0.8g/ml, twice every day) by intragastric administration separately after the end of 8th week, meanwhile the normal and model group were separately given life drinking water by the same means. All the rats were sacrificed after 4,8,12 weeks. Since the abdominal aortic blood, quickly remove the liver, according to conventional preparation of serum, liver homogenate, and to retain a small amount of liver tissue cryopreservation in-70℃.
2. Detect indicators:measured body weight, body length, wet weight of liver, and calculate the liver index (liver wet weight/body weight×100%), Lee's index (0.33 weight/body length×1000) and other physical indicators. Automatic biochemical analyzer determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood lipids content, light microscope to assess steatosis, inflammatory activity and fibrosis; The levels of Bcl-2 and Bax expression were determined by Western Blot.
Results:1. The liver lobules of normal group rats structural integrity, liver cord around the central venous actinomorphous distribution, cells were polygonal, various biochemical indices were within the normal range; the rates of Bcl-2 and Bax keep the balance.
2. After 4 weeks, the model rats exhibited mild steatosis, no inflammatory cell infiltrationand and no fibrosis; increase of aminotransferase; compared with normal group, the expression of Bax protein was more pronounced, while the expression of Bcl-2 was decreased, Bcl-2/Bax was also decreased significantly (P<0.01).
3. After 8 weeks, control with the N2 group, model group M2 rats, simple fatty liver were observed, no inflammatory cell infiltration and fibrosis S1; serum transaminase levels were significantly higher than normal, TC, TG, ALT, AST; liver Bax expression than the control group increased significantly, while Bcl-2, Bcl-2/Bax decreased (P<0.01).
4. After 12 weeks, severe steatohepatitis with fibrosis S2 of M3 rats; compared with normal group, the level of aminotransferase and blood fat were increased in model group; liver Bax expression than the control group increased significantly, while Bcl-2, Bcl-2/Bax decreased; compared with M3 group, the treated group every biochemical manifestation shew improvement significantly, the expression of Bax in liver was decreased, Bcl-2 and Bcl-2/Bax increased; but still lower than normal group.
Conclusion:1. High-fat and high-cholesterol diet can successfully copy the NAFLD rat model; lipid metabolism disorders, inflammatory injury and apoptosis are closely related to the occurrence of NAFLD.
2. Bcl-2 and Bax can be expressed in normal rat liver, the expression of Bax protein was more pronounced from 4 week and continued to rise until 8 week and 12 week in the model group, compared with the normal group the expression of Bcl-2 was decreased, Bcl-2/Bax in the model group, particularly at 12 week decreased in a time-dependent manner.
3. Jiangzhi Yigan granule has a good anti-therapeutic effects for combat NAFLD. Its molecular mechanism may be partly through up-regulating the expression of Bcl-2 of the live, down-regulating Bax and keeping the balance of Bcl-2 and Bax.
引文
[1]Bjornsson E, Angulo P. Non-alcoholic fatty liver disease. Scand J Gastroenterol,2007, 42(9):1023-1030.
[2]Duvnjak M, Lerotic I, Barsic N, et al. Pathogenesis and management issues for non-alcoholic fatty liver disease. World J Gastroenterol,2007,13(34):4539-4550.
[3]Ong JP, Younossi ZM. Epidemiology and natural history of NAFLD and NASH. Clin Liver Dis.2007,11(1):1-16.
[4]Farrell GC, Larter CZ. Nonalcoholic fatty liver disease:from steatosis to cirrhosis. Hepatology.2006,43(2 Suppl 1):S99-S112
[5]Day CP, James OF. Steatohepatitis:a tale of two "hits"? Gastroenterology,1998,114(4): 842-845.
[6]Wanless IR, Shiota K. The pathogenesis of nonalcoholic steatohepatitis and other fatty liver diseases:a four-step model including the role of lipid release and hepatic venular obstruction in the progression to cirrhosis.Semin Liver Dis.2004,24(l):99-106.
[7]Wieckowska A, Zein NN, Yerian LM, et al. In vivo assessment of liver cell apoptosis as a novel biomarker of disease severity in nonalcoholic fatty liver disease. Hepatology, 2006,44(1):27-33.
[8]Day CP. Natural history of NAFLD:remarkably benign in the absence of cirrhosis [J]. Gastroenterology,2005,129(1):375-378.
[9]Harper N, Hughes M, MacFarlane M, et al. Fas-associated death domain protein and caspase-8 are not recruited to the tumor necrosis factor receptor 1 signaling complex during tumor necrosis factor-induced apoptosis. J Biol Chem,2003,278(28):25534-25541.
[10]Jaeschke H, Gores GJ, Cederbaum Al, et al. Mechanisms of hepatotoxicity. Toxicol Sci.2002,65(2):166-176.
[11]Baskin-Bey ES, Gores GJ. Death by association:BH3 domain-only proteins and liver injury. Am J Physiol Gastrointest Liver Physiol,2005,289(6):987-990.
[12]Tsujimoto Y, Cossman J, Jaffe E, et al. Involvement of the bcl-2 gene in human follicular lymphoma [J]. Science,1985,228 (4706):1440-1443.
[13]Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homology, Bax, that accelerates programmed cell death [J]. Cell,1993,74 (4):609-619.
[14]曹智刚,袁守军.抑制Bax基因表达与细胞保护的研究进展.中国临床药理学与治疗学,2003,8(3)：245-248.
[15]付永锋,樊廷俊.Bcl-2家族蛋白与细胞凋亡.生物化学与生物物理学报,2002,34 (4):389-394.
[16]Rogerio F, Jordao H Jr, Vieira AS, et al. Bax and Bcl-2 expression and TUNEL labeling in lumbar enlargement of neonatal rats after sciatic axotomy and melatonin treatment. Brain Res,2006,1112(1):80-90.
[17]Higuchi M, Aggarwal BB, Yeh ET. Activation of CPP32-like protease in tumor necrosis factor-induced apoptosis is dependent on mitochondrial function. J Clin Invest,1997,99(7):1751-1758.
[18]Angulo P. Current best treatment for non-alcoholic fatty liver disease. Expert Opin Pharmacother,2003,4(5):611-623.
[19]殷果华,殷云勤,赵和平,等.降脂益肝冲剂治疗脂肪肝的实验观察.中华实验和临床病毒学杂志,2000,14(2)：193-194.
[20]赵睿峰.降脂益肝冲剂对NASH的影响.[硕士学位论文]山西：山西医科大学,2006.
[21]戴林,邓彬,白成,等.吡格列酮对大鼠非酒精性脂肪性肝炎干预的实验研究.胃肠病学和肝病学杂志,2004,13(5)：482-484.
[22]徐叔云,卞如廉,陈修.药理实验方法学[M].第2版.北京：人民卫生出版社.1991,178-179.
[23]栗凤霞.复方中药治疗NAFLD的实验研究PPA-a及其配体作用.[硕士学位论文].天津：天津医科大学,2004.
[24]Knodell RG, Ishak KG, Black WC, et al. Formulation and application of a numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis. Hepatol,1981,1(5):431-435.
[25]王泰龄,刘霞,周元平,等.慢性肝炎炎症活动度及纤维化程度计分方案.中华肝脏病杂志,1998,6(4)：195-197.
[26]中华医学会肝脏病学分会脂肪肝和酒精性肝病学组.非酒精性脂肪性肝病诊断标准.中华肝脏病杂志.2003,11(2)：71.
[27]中华医学会传染病与寄生虫病学分会、肝病学分会.病毒性肝炎防治方案.中华内科杂志,2001,40(1)：62-68
[28]Lieber CS, Leo MA, Mak KM, et al. Model of nonalcoholic steatohepatitis. Am J Clin Nutr.2004,79(3):502-509.
[29]钟岚,范建高,王国良,等.非酒精性脂肪性肝炎发病机制的实验研究.中华消化杂志,2001,21(8)：481-484.
[30]Kerr JF, Wyllie AH, Currie AR. Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972,26(4):239-257.
[31]Jaeschke H, Bajt ML. Regulation of apoptotic signaling pathways in hepatocytes in vivo. Hepatology,2003,37(4):942-945.
[32]Kohlhaas SL, Craxton A, Sun XM, et al. Receptor-mediated endocytosis is not required for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. J Biol Chem.2007,282(17):12831-12841.
[33]Jaeschke H, Gores GJ, Cederbaum AI, et al. Mechanisms of hepatotoxicity. Toxicol Sci.2002,65(2):166-176.
[34]Harper N, Hughes M, MacFarlane M, et al. Fas-associated death domain protein and caspase-8 are not recruited to the tumor necrosis factor receptor 1 signaling complex during tumor necrosis factor-induced apoptosis. J Biol Chem,2003,278(28):25534-25541.
[35]Yin XM, Ding WX. Death receptor activation-induced hepatocyte apoptosis and liver injury. Curr Mol Med,2003,3(6):491-508.
[36]Ribeiro PS, Cortez-Pinto H, Sola S, et al. Hepatocyte apoptosis, expression of death receptors, and activation of NF-kappaB in the liver of nonalcoholic and alcoholic steatohepatitis patients. Am J Gastroenterol,2004,99(9):1708-1717.
[37]Hatano E. Tumor necrosis factor signaling in hepatocyte apoptosis. J Gastroenterol Hepatol,2007,22 Suppl 1:S43-S44.
[38]Chen X, Ding WX, Ni HM, et al. Bid-independent mitochondrial activation in tumor necrosis factor alpha-induced apoptosis and liver injury. Mol Cell Biol,2007,27(2): 541-553.
[39]Schinoni MI, Parana R. Apoptosis and progression of hepatic fibrosis in liver diseases. Acta Gastroenterol Latinoam,2006,36(4):211-217.
[40]Hsu H, Shu HB, Pan MG, et al. TRADD-TRAF2 and TRADD-FADD interactions define two distinct TNF receptor 1 signal transduction pathways. Cell,1996,84(2): 299-308.
[41]Ding D, Jiang H, Wang P, et al. Cell death after co-administration of cisplatin and ethacrynic acid. Hear Res.2007,226(1-2):129-139.
[42]Chen D, Texada DE, Duggan C, et al. Surface calreticulin mediates muramyl dipeptide-induced apoptosis in RK13 cells. J Biol Chem.2005,280(23):22425-22436.
[43]Dorscheid DR, Wojcik KR, Sun S, et al. Apoptosis of airway epithelial cells induced by corticosteroids. Am J Respir Crit Care Med,2001,164(10):1939-1947.
[44]White SR, Dorscheid DR. Corticosteroid-induced apoptosis of airway epithelium:a potential mechanism for chronic airway epithelial damage in asthma. Chest, 2002,122(6):278-284.
[45]Zamzami N, Kroemer G. Apopotosis:mitochondrial membrane permeabilization-the (w)hole story?. Curr Biol,2003,13(2):71-73.
[46]Strasser A, O'Connor L, Dixit VM. Apopotosis signaling. Annu Rev Biochem,2000, 69(1):217-245.
[47]Wei MC, Zong WX, Cheng EH, et al. Proapopptotic BAX and BAK:a requisite gateway to mitochondrial dysfunction and death. Science,2001,292(5517):727-730.
[48]Boisvieux-Ulrich E, Sourdeval M, Marano F. CD437, a synthetic retinoid, induces apoptosis in human respiratory epithelial cells via caspase-independent mitochondrial and caspase-8-dependent pathways both up-regulated by JNK signaling pathway[J]. Exp Cell Res,2005,307 (1):76-90.
[49]Rust C, Gores GJ. Apoptosis and liver disease. Am J Med,2000,108(7):567-574.
[50]Hannun YA. Apoptosis and the dilemma of cancer chemotherapy. Blood,1997,89(6): 1845-1853.
[51]Gao Z, Tian Y, Wang J, et al. A dimeric Smac/diablo peptide directly relieves caspase-3 inhibition by XIAP. Dynamic and-cooperative regulation of XIAP by Smac/Diablo. J Biol Chem,2007,282(42):30718-30727.
[52]Chen DF, Wang CH. The relationship between the opening of mitochondrial permeability transition pores of cultured hepatocytes with their apoptoses in a non-alcoholic fatty liver disease model. Zhonghua Gan Zang Bing Za Zhi,2007,15(11): 837-839.
[53]Gomez-Lechon MJ, Donato MT, Martinez-Romero A, et al. A human hepatocellular in vitro model to investigate steatosis. Chem Biol Interact,2007,165(2):106-116.
[54]Ji J, Zhang L, Wang P, et al. Saturated free fatty acid, palmitic acid, induces apoptosis in fetal hepatocytes in culture. Exp Toxicol Pathol,2005,56:369-376.
[55]Mitsuyoshi H, Itoh Y, Okanoue T. Role of oxidative stress in non-alcoholic steatohepatitis. Nippon Rinsho,2006,64(6):1077-1082.
[56]Panasiuk A, Dzieciol J, Panasiuk B, et al. Expression of p53, Bax and Bcl-2 proteins in hepatocytes in non-alcoholic fatty liver disease. World J Gastroenterol,2006,12(38): 6198-6202.
[57]Ramalho RM, Cortez-Pinto H, Castro RE, et al. Apoptosis and Bcl-2 expression in the livers of patients with steatohepatitis. Eur J Gastroenterol Hepatol,2006,18(1):21-29.
[58]南月敏,乔梁,于君,等.Fas及其配体诱导非酒精性脂肪性肝炎肝细胞凋亡.中华消化杂志,2006,26(12)：841-842.
[59]Abdeen MB, Chowdhury NA, Hayden MR,et al. Nonalcoholic steatohepatitis and the cardiometabolic syndrome. J Cardiometab Syndr,2006,1(1):36-40.
[60]范建高,钟岚,王国良,等.肥胖、高脂血症性脂肪性肝炎细胞凋亡的实验研究. 中华消化杂志,2000,20(6)：421-422.
[60]Mitchell C, Mallet V, Guidotti JE, et al. Experimental modulation of apoptosis: physiopathological and therapeutic targets. J Soc Biol,2005,199(3):243-246.
[62]Shimizu I, Kohno N, Tamaki K, et al. Female hepatology:favorable role of estrogen in chronic liver disease with hepatitis B virus infection. World J Gastroenterol,2007, 13(32):4295-4305.
[63]Feldstein AE, Canbay A, Angulo P, et al. Hepatocyte apoptosis and fas expression are prominent features of human nonalcoholic steatohepatitis. Gastroenterology,2003, 125(2):437-443.
[64]Nan YM, Wu WJ, Yao XX, et al. The role of apoptosis and the related genes in non-alcoholic steatohepatitis. Zhonghua Gan Zang Bing Za Zhi,2007,15(1):41-46.
[65]Carter-Kent C, Zein NN, Feldstein AE. Cytokines in the pathogenesis of fatty liver and disease progression to steatohepatitis:implications for treatment. Am J Gastroenterol, 2008,103(4):1036-1042.
[66]Marra F, Gastaldelli A, Svegliati Baroni G, et al. Molecular basis and mechanisms of progression of non-alcoholic steatohepatitis. Trends Mol Med,2008,14(2):72-81.
[67]Canbay A, Taimr P, Torok N, et al. Apoptotic body engulfment by a human stellate cell line is profibrogenic. Lab Invest,2003,83(5):655-663.
[68]Canbay A, Feldstein AE, Higuchi H, et al. Kupffer cell engulfment of apoptotic bodies stimulates death ligand and cytokine expression. Hepatology,2003,38(5):1188-1198.
[69]Canbay A, Kip SN, Kahraman A, et al. Apoptosis and fibrosis in non-alcoholic fatty liver disease. Turk J Gastroenterol,2005,16(1):1-6.
[70]Roset R, Ortet L, Gil-Gomez G. Role of Bcl-2 family members on apoptosis:what we have learned from knock-out mice. Front Biosci,2007,12(20):4722-4730.
[71]Hoetelmans R, Van Slooten HJ, Keijzer R, et al. Bcl-2 and Bax proteins are present in interphase nuclei of mammalian cells. Cell Death Differ,2000,7(4):384-392.
[72]Korsmeyer SJ, Shutter JR, Veis DJ, et al. Bcl-2/Bax:a rheostat that regulates an anti-oxidant pathway and cell death. Semin Cancer Biol,1993,4(6):327-332.
[73]Mathai JP, Germain M, Shore GC. BH3-only BIK regulates BAX, BAK-dependent release of Ca2+from endoplasmic reticulum stores and mitochondrial apoptosis during stress-induced cell death. J Biol Chem,2005,280(25):23829-23836.
[74]Chami M, Prandini A, Campanella M, et al. Bcl-2 and Bax exert opposing effects on Ca2+ signaling, which do not depend on their putative pore-forming region. J Biol Chem, 2004,279(52):54581-54589.
[75]Voehringer DW, Meyn RE. Redox aspects of Bcl-2 function. Antioxid Redox Signal, 2000,2(3):537-550.
[76]Metrailler-Ruchonnet I, Pagano A, Carnesecchi S, et al. Bcl-2 protects against hyperoxia-induced apoptosis through inhibition of the mitochondria-dependent pathway. Free Radic Biol Med,2007,42 (7):1062-1074.
[77]Ellerby LM, Ellerby HM, Park SM, et al. Shift of the cellular oxidation-reduction potential in neural cells expressing Bcl-2. J Neurochem,1996,67(3):1259-1267.
[78]Brooks C, Dong Z. Regulation of mitochondrial morphological dynamics during apoptosis by Bcl-2 family proteins:a key in Bak?. Cell Cycle,2007,6(24):3043-3047.
[79]樊丽琳,陈东风,胡辂,等.bcl-2、bax在大鼠非酒精性脂肪性肝病中的表达及意义.重庆医学,2007,36(8)：692-694.
[80]Vaux DL, Cory S, Adams JM. Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pro-B cells. Nature,1998,335 (6189):440-442.
[81]Sato H, Ozawa K, Iwata S, et al. Role of bcl-2 mRNA in homeostatic proliferation in circulating T-cells in human liver transplant patients after T-cell depletion. J Surg Res, 2005,127(2):123-130.
[82]Bermudez Y, Erasso D, Johnson NC, et al. Telomerase confers resistance to caspase-mediated apoptosis. Clin Interv Aging,2006,1(2):155-167.
[83]Malhi H, Bronk SF, Werneburg NW, et al. Free fatty acids induce JNK-dependent hepatocyte lipoapoptosis. J Biol Chem,2006,281(17):12093-12101.
[84]Fletcher JI, Huang DC. Controlling the cell death mediators Bax and Bak:puzzles and conundrums. Cell Cycle,2008,7(1):39-44.
[85]Debatin KM. Chronic lymphocytic leukemia:keeping cell death at bay. Cell,2007, 129(5):853-855.
[86]Wong CK, Chen Z, So KL, et al. Polycomb group protein RING IB is a direct substrate of Caspases-3 and-9. Biochim Biophys Acta,2007,1773(6):844-852.
[87]Bao Q, Lu W, Rabinowitz JD, et al. Calcium blocks formation of apoptosome by preventing nucleotide exchange in Apaf-1. Mol Cell,2007,25(2):181-192.
[88]张静,赵景民,周光德,等.非酒精性脂肪性肝炎肝细胞凋亡及凋亡相关蛋白的表达.解放军医学杂志,2008,33(5)：521-523.
[89]Tsukahara S, Yamamoto S, Tin-Tin-Win-Shwe, et al. Inhalation of low-level formaldehyde increases the bcl-2/bax expression ratio in the hippocampus of immunologically sensitized mice. Neuroimmunomodulation,2006,13(2):63-68.
[90]殷果华,殷云勤,赵和平,等.降脂益肝冲剂治疗脂肪肝的实验观察.中华实验和临床病毒学杂志,2000,14(2)：193-194.
[91]胡义扬,刘平,刘成,等.丹参提取物对CC1-4和DMN诱导的大鼠肝纤维化的影响. 上海中医药杂志,1999(10)：7-10.
[92]洪学智.泽泻治疗非酒精性脂肪肝病药效及作用机理研究.[硕士学位论文].浙江大学,2006.
[93]李英,王晓素.脂肪肝的中医药治疗现状.甘肃中医,2006,19(3)：35-37.
[94]王德山,肖玉芳,董朝晖,等.枸杞抗实验性高血脂肝脂量效关系及毒性研究.辽宁中医杂志,1997,24(12)：567-568.
[95]李续娥,郭宝江.决明子蛋白质和葸醌苷对高脂血症大鼠血脂的影响.中国临床康复,2004,8(18)：3694-3695.
[96]李贵海,孙敬勇,张希林,等.山楂降血脂有效成分的实验研究.中草药,2002,33(1)：50-52.
[97]Bjorntorp P. Metabolic difference between visceral fat and subcutaneous abdominal fat. Diabetes Metab,2000,26(Suppl 3):10-12.
[1]Wieckowska A, Zein NN, Yerian LM, et al. In vivo assessment of liver cell apoptosis as a novel biomarker of disease severity in nonalcoholic fatty liver disease[J]. Hepatology, 2006,44(1):27-33.
[2]Tsukahara S, Yamamoto S, Tin-Tin-Win-Shwe, et al. Inhalation of low-level formaldehyde increases the Bcl-2/Bax expression ratio in the hippocampus of immunologically sensitized mice[J]. Neuroimmunomodulation,2006,13(2):63-68.
[3]Tsujimoto Y, Cossman J, Jaffe E, et al. Involvement of the bcl-2 gene in human follicular lymphoma[J]. Science,1985,228 (4706):1440-1443.
[4]Roset R, Ortet L, Gil-Gomez G. Role of Bcl-2 family members on apoptosis:what we have learned from knock-out mice [J]. Front Biosci,2007,12 (20):4722-4730.
[5]Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death[J]. Cell,1993,74 (4):609-619.
[6]Hoetelmans R, Van Slooten HJ, Keijzer R, et al. Bcl-2 and Bax proteins are present in interphasenuclei of mammalian cells[J]. Cell Death Differ,2000,7(4):384-392.
[7]陈东风,王春华.体外非酒精性脂肪肝细胞模型线粒体通透转换孔的变化及其与细胞凋亡的关系[J].中华肝脏病杂志,2007,15(11)：837-839.
[8]Dorscheid DR, Wojcik KR, Sun S, et al. Apoptosis of airway epithelial cells induced by corticosteroids[J]. Am J Respir Crit Care Med,2001,164(10):1939-1947.
[9]White SR, Dorscheid DR. Corticosteroid-induced apoptosis of airway epithelium:a potential mechanism for chronic airway epithelial damage in asthma[J]. Chest, 2002,122(6):278-284.
[10]Zamzami N, Kroemer G. Apopotosis:mitochondrial membrane permeabilization-the (w)hole story? [J]. Curr Biol,2003,13(2):71-73.
[11]Strasser A, O'Connor L, Dixit VM, et al. Apopotosis signaling[J]. Annu Rev Biochem, 2000,69 (1):217-245.
[12]Wei MC, Zong WX, Cheng EH, et al. Proapopptotic BAX and BAK:a requisite gateway to mitochondrial dysfunction and death[J]. Science,2001,292(5517):727-730.
[13]Boisvieux-Ulrich E, Sourdeval M, Marano F. CD437, a synthetic retinoid, induces apoptosis in human respiratory epithelial cells via caspase-independent mitochondrial and caspase-8-dependent pathways both up-regulated by JNK signaling pathway[J]. Exp Cell Res,2005,307(1):76-90.
[14]Rust C, Gores GJ. Apoptosis and liver disease[J]. Am J Med,2000,108(7):567-574.
[15]Hannun YA. Apoptosis and the dilemma of cancer chemotherapy[J]. Blood,1997, 89(6):1845-1853.
[16]Mathai JP, Germain M, Shore GC. BH3-only BIK regulates BAX, BAK-dependent release of Ca2+from endoplasmic reticulum stores and mitochondrial apoptosis during stress-induced cell death[J]. J Biol Chem,2005,280(25):23829-23836.
[17]Chami M, Prandini A, Campanella M, et al. Bcl-2 and Bax exert opposing effects on Ca2+ signaling, which do not depend on their putative pore-forming region[J]. J Biol Chem,2004,279(52):54581-54589.
[18]Voehringer DW, Meyn RE. Redox aspects of Bcl-2 function[J]. Antioxid Redox Signal, 2000,2(3):537-550.
[19]Metrailler-Ruchonnet I, Pagano A, Carnesecchi S, et al. Bcl-2 protects against hyperoxia-induced apoptosis through inhibition of the mitochondria-dependent pathway[J]. Free Radic BioIMed,2007,42 (7):1062-1074.
[20]Ellerby LM, Ellerby HM, Park SM, et al. Shift of the cellular oxidation-reduction potential in neural cells expressing Bcl-2[J]. J Neurochem,1996,67(3):1259-1267.
[21]Brooks C, Dong Z. Regulation of mitochondrial morphological dynamics during apoptosis by Bcl-2 family proteins:a key in Bak?[J]. Cell Cycle,2007,6 (24): 3043-3047.
[22]樊丽琳,陈东风,胡辂,等.bcl-2、bax在大鼠非酒精性脂肪性肝病中的表达及意义[J].重庆医学,2007,36(8)：692-694.
[23]Vaux DL, Cory S, Adams JM. Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pro-B cells[J]. Nature,1998,335 (6189): 440-442.
[24]Sato H, Ozawa K, Iwata S, et al. Role of bcl-2 mRNA in homeostatic proliferation in circulating T-cells in human liver transplant patients after T-cell depletion[J]. J Surg Res,2005,127(2):123-130.
[25]Bermudez Y, Erasso D, Johnson NC, et al. Telomerase confers resistance to caspase-mediated apoptosis[J]. Clin Interv Aging,2006,1(2):155-167.
[26]Malhi H, Bronk SF, Werneburg NW, et al. Free fatty acids induce JNK-dependent hepatocyte lipoapoptosis[J]. J Biol Chem,2006,281(17):12093-12101.
[27]Fletcher JI, Huang DC. Controlling the cell death mediators Bax and Bak:puzzles and conundrums[J]. Cell Cycle,2008,7(1):39-44.
[28]Debatin KM. Chronic lymphocytic leukemia:keeping cell death at bay[J]. Cell,2007, 129(5):853-855.
[29]Wong CK, Chen Z, So KL, et al. Polycomb group protein RING1B is a direct substrate of Caspases-3 and-9[J]. Biochim Biophys Acta,2007,1773(6):844-852.
[30]Bao Q, Lu W, Rabinowitz JD, et al. Calcium blocks formation of apoptosome by preventing nucleotide exchange in Apaf-1[J]. Mol Cell,2007,25(2):181-192.
[31]Rogerio F, Jordao H Jr, Vieira AS, et al. Bax and Bcl-2 expression and TUNEL labeling in lumbar enlargement of neonatal rats after sciatic axotomy and melatonin treatment[J]. Brain Res,2006,1112(1):80-90.
[32]张静,赵景民,周光德,等.非酒精性脂肪性肝炎肝细胞凋亡及凋亡相关蛋白的表达[J].解放军医学杂志,2008,33(5)：521-523.
[33]Higuchi M, Aggarwal BB, Yeh ET. Activation of CPP32-like protease in tumor necrosis factor-induced apoptosis is dependent on mitochondrial function[J]. J Clin Invest,1997,99 (7):1751-1758.
[2]Duvnjak M, Lerotic I, Barsic N, et al. Pathogenesis and management issues for non-alcoholic fatty liver disease. World J Gastroenterol,2007,13(34):4539-4550.
[3]Ong JP, Younossi ZM. Epidemiology and natural history of NAFLD and NASH. Clin Liver Dis.2007,11(1):1-16.
[4]Farrell GC, Larter CZ. Nonalcoholic fatty liver disease:from steatosis to cirrhosis. Hepatology.2006,43(2 Suppl 1):S99-S112
[5]Day CP, James OF. Steatohepatitis:a tale of two "hits"? Gastroenterology,1998,114(4): 842-845.
[6]Wanless IR, Shiota K. The pathogenesis of nonalcoholic steatohepatitis and other fatty liver diseases:a four-step model including the role of lipid release and hepatic venular obstruction in the progression to cirrhosis.Semin Liver Dis.2004,24(l):99-106.
[7]Wieckowska A, Zein NN, Yerian LM, et al. In vivo assessment of liver cell apoptosis as a novel biomarker of disease severity in nonalcoholic fatty liver disease. Hepatology, 2006,44(1):27-33.
[8]Day CP. Natural history of NAFLD:remarkably benign in the absence of cirrhosis [J]. Gastroenterology,2005,129(1):375-378.
[9]Harper N, Hughes M, MacFarlane M, et al. Fas-associated death domain protein and caspase-8 are not recruited to the tumor necrosis factor receptor 1 signaling complex during tumor necrosis factor-induced apoptosis. J Biol Chem,2003,278(28):25534-25541.
[10]Jaeschke H, Gores GJ, Cederbaum Al, et al. Mechanisms of hepatotoxicity. Toxicol Sci.2002,65(2):166-176.
[11]Baskin-Bey ES, Gores GJ. Death by association:BH3 domain-only proteins and liver injury. Am J Physiol Gastrointest Liver Physiol,2005,289(6):987-990.
[12]Tsujimoto Y, Cossman J, Jaffe E, et al. Involvement of the bcl-2 gene in human follicular lymphoma [J]. Science,1985,228 (4706):1440-1443.
[13]Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homology, Bax, that accelerates programmed cell death [J]. Cell,1993,74 (4):609-619.
[14]曹智刚,袁守军.抑制Bax基因表达与细胞保护的研究进展.中国临床药理学与治疗学,2003,8(3)：245-248.
[15]付永锋,樊廷俊.Bcl-2家族蛋白与细胞凋亡.生物化学与生物物理学报,2002,34 (4):389-394.
[16]Rogerio F, Jordao H Jr, Vieira AS, et al. Bax and Bcl-2 expression and TUNEL labeling in lumbar enlargement of neonatal rats after sciatic axotomy and melatonin treatment. Brain Res,2006,1112(1):80-90.
[17]Higuchi M, Aggarwal BB, Yeh ET. Activation of CPP32-like protease in tumor necrosis factor-induced apoptosis is dependent on mitochondrial function. J Clin Invest,1997,99(7):1751-1758.
[18]Angulo P. Current best treatment for non-alcoholic fatty liver disease. Expert Opin Pharmacother,2003,4(5):611-623.
[19]殷果华,殷云勤,赵和平,等.降脂益肝冲剂治疗脂肪肝的实验观察.中华实验和临床病毒学杂志,2000,14(2)：193-194.
[20]赵睿峰.降脂益肝冲剂对NASH的影响.[硕士学位论文]山西：山西医科大学,2006.
[21]戴林,邓彬,白成,等.吡格列酮对大鼠非酒精性脂肪性肝炎干预的实验研究.胃肠病学和肝病学杂志,2004,13(5)：482-484.
[22]徐叔云,卞如廉,陈修.药理实验方法学[M].第2版.北京：人民卫生出版社.1991,178-179.
[23]栗凤霞.复方中药治疗NAFLD的实验研究PPA-a及其配体作用.[硕士学位论文].天津：天津医科大学,2004.
[24]Knodell RG, Ishak KG, Black WC, et al. Formulation and application of a numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis. Hepatol,1981,1(5):431-435.
[25]王泰龄,刘霞,周元平,等.慢性肝炎炎症活动度及纤维化程度计分方案.中华肝脏病杂志,1998,6(4)：195-197.
[26]中华医学会肝脏病学分会脂肪肝和酒精性肝病学组.非酒精性脂肪性肝病诊断标准.中华肝脏病杂志.2003,11(2)：71.
[27]中华医学会传染病与寄生虫病学分会、肝病学分会.病毒性肝炎防治方案.中华内科杂志,2001,40(1)：62-68
[28]Lieber CS, Leo MA, Mak KM, et al. Model of nonalcoholic steatohepatitis. Am J Clin Nutr.2004,79(3):502-509.
[29]钟岚,范建高,王国良,等.非酒精性脂肪性肝炎发病机制的实验研究.中华消化杂志,2001,21(8)：481-484.
[30]Kerr JF, Wyllie AH, Currie AR. Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972,26(4):239-257.
[31]Jaeschke H, Bajt ML. Regulation of apoptotic signaling pathways in hepatocytes in vivo. Hepatology,2003,37(4):942-945.
[32]Kohlhaas SL, Craxton A, Sun XM, et al. Receptor-mediated endocytosis is not required for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. J Biol Chem.2007,282(17):12831-12841.
[33]Jaeschke H, Gores GJ, Cederbaum AI, et al. Mechanisms of hepatotoxicity. Toxicol Sci.2002,65(2):166-176.
[34]Harper N, Hughes M, MacFarlane M, et al. Fas-associated death domain protein and caspase-8 are not recruited to the tumor necrosis factor receptor 1 signaling complex during tumor necrosis factor-induced apoptosis. J Biol Chem,2003,278(28):25534-25541.
[35]Yin XM, Ding WX. Death receptor activation-induced hepatocyte apoptosis and liver injury. Curr Mol Med,2003,3(6):491-508.
[36]Ribeiro PS, Cortez-Pinto H, Sola S, et al. Hepatocyte apoptosis, expression of death receptors, and activation of NF-kappaB in the liver of nonalcoholic and alcoholic steatohepatitis patients. Am J Gastroenterol,2004,99(9):1708-1717.
[37]Hatano E. Tumor necrosis factor signaling in hepatocyte apoptosis. J Gastroenterol Hepatol,2007,22 Suppl 1:S43-S44.
[38]Chen X, Ding WX, Ni HM, et al. Bid-independent mitochondrial activation in tumor necrosis factor alpha-induced apoptosis and liver injury. Mol Cell Biol,2007,27(2): 541-553.
[39]Schinoni MI, Parana R. Apoptosis and progression of hepatic fibrosis in liver diseases. Acta Gastroenterol Latinoam,2006,36(4):211-217.
[40]Hsu H, Shu HB, Pan MG, et al. TRADD-TRAF2 and TRADD-FADD interactions define two distinct TNF receptor 1 signal transduction pathways. Cell,1996,84(2): 299-308.
[41]Ding D, Jiang H, Wang P, et al. Cell death after co-administration of cisplatin and ethacrynic acid. Hear Res.2007,226(1-2):129-139.
[42]Chen D, Texada DE, Duggan C, et al. Surface calreticulin mediates muramyl dipeptide-induced apoptosis in RK13 cells. J Biol Chem.2005,280(23):22425-22436.
[43]Dorscheid DR, Wojcik KR, Sun S, et al. Apoptosis of airway epithelial cells induced by corticosteroids. Am J Respir Crit Care Med,2001,164(10):1939-1947.
[44]White SR, Dorscheid DR. Corticosteroid-induced apoptosis of airway epithelium:a potential mechanism for chronic airway epithelial damage in asthma. Chest, 2002,122(6):278-284.
[45]Zamzami N, Kroemer G. Apopotosis:mitochondrial membrane permeabilization-the (w)hole story?. Curr Biol,2003,13(2):71-73.
[46]Strasser A, O'Connor L, Dixit VM. Apopotosis signaling. Annu Rev Biochem,2000, 69(1):217-245.
[47]Wei MC, Zong WX, Cheng EH, et al. Proapopptotic BAX and BAK:a requisite gateway to mitochondrial dysfunction and death. Science,2001,292(5517):727-730.
[48]Boisvieux-Ulrich E, Sourdeval M, Marano F. CD437, a synthetic retinoid, induces apoptosis in human respiratory epithelial cells via caspase-independent mitochondrial and caspase-8-dependent pathways both up-regulated by JNK signaling pathway[J]. Exp Cell Res,2005,307 (1):76-90.
[49]Rust C, Gores GJ. Apoptosis and liver disease. Am J Med,2000,108(7):567-574.
[50]Hannun YA. Apoptosis and the dilemma of cancer chemotherapy. Blood,1997,89(6): 1845-1853.
[51]Gao Z, Tian Y, Wang J, et al. A dimeric Smac/diablo peptide directly relieves caspase-3 inhibition by XIAP. Dynamic and-cooperative regulation of XIAP by Smac/Diablo. J Biol Chem,2007,282(42):30718-30727.
[52]Chen DF, Wang CH. The relationship between the opening of mitochondrial permeability transition pores of cultured hepatocytes with their apoptoses in a non-alcoholic fatty liver disease model. Zhonghua Gan Zang Bing Za Zhi,2007,15(11): 837-839.
[53]Gomez-Lechon MJ, Donato MT, Martinez-Romero A, et al. A human hepatocellular in vitro model to investigate steatosis. Chem Biol Interact,2007,165(2):106-116.
[54]Ji J, Zhang L, Wang P, et al. Saturated free fatty acid, palmitic acid, induces apoptosis in fetal hepatocytes in culture. Exp Toxicol Pathol,2005,56:369-376.
[55]Mitsuyoshi H, Itoh Y, Okanoue T. Role of oxidative stress in non-alcoholic steatohepatitis. Nippon Rinsho,2006,64(6):1077-1082.
[56]Panasiuk A, Dzieciol J, Panasiuk B, et al. Expression of p53, Bax and Bcl-2 proteins in hepatocytes in non-alcoholic fatty liver disease. World J Gastroenterol,2006,12(38): 6198-6202.
[57]Ramalho RM, Cortez-Pinto H, Castro RE, et al. Apoptosis and Bcl-2 expression in the livers of patients with steatohepatitis. Eur J Gastroenterol Hepatol,2006,18(1):21-29.
[58]南月敏,乔梁,于君,等.Fas及其配体诱导非酒精性脂肪性肝炎肝细胞凋亡.中华消化杂志,2006,26(12)：841-842.
[59]Abdeen MB, Chowdhury NA, Hayden MR,et al. Nonalcoholic steatohepatitis and the cardiometabolic syndrome. J Cardiometab Syndr,2006,1(1):36-40.
[60]范建高,钟岚,王国良,等.肥胖、高脂血症性脂肪性肝炎细胞凋亡的实验研究. 中华消化杂志,2000,20(6)：421-422.
[60]Mitchell C, Mallet V, Guidotti JE, et al. Experimental modulation of apoptosis: physiopathological and therapeutic targets. J Soc Biol,2005,199(3):243-246.
[62]Shimizu I, Kohno N, Tamaki K, et al. Female hepatology:favorable role of estrogen in chronic liver disease with hepatitis B virus infection. World J Gastroenterol,2007, 13(32):4295-4305.
[63]Feldstein AE, Canbay A, Angulo P, et al. Hepatocyte apoptosis and fas expression are prominent features of human nonalcoholic steatohepatitis. Gastroenterology,2003, 125(2):437-443.
[64]Nan YM, Wu WJ, Yao XX, et al. The role of apoptosis and the related genes in non-alcoholic steatohepatitis. Zhonghua Gan Zang Bing Za Zhi,2007,15(1):41-46.
[65]Carter-Kent C, Zein NN, Feldstein AE. Cytokines in the pathogenesis of fatty liver and disease progression to steatohepatitis:implications for treatment. Am J Gastroenterol, 2008,103(4):1036-1042.
[66]Marra F, Gastaldelli A, Svegliati Baroni G, et al. Molecular basis and mechanisms of progression of non-alcoholic steatohepatitis. Trends Mol Med,2008,14(2):72-81.
[67]Canbay A, Taimr P, Torok N, et al. Apoptotic body engulfment by a human stellate cell line is profibrogenic. Lab Invest,2003,83(5):655-663.
[68]Canbay A, Feldstein AE, Higuchi H, et al. Kupffer cell engulfment of apoptotic bodies stimulates death ligand and cytokine expression. Hepatology,2003,38(5):1188-1198.
[69]Canbay A, Kip SN, Kahraman A, et al. Apoptosis and fibrosis in non-alcoholic fatty liver disease. Turk J Gastroenterol,2005,16(1):1-6.
[70]Roset R, Ortet L, Gil-Gomez G. Role of Bcl-2 family members on apoptosis:what we have learned from knock-out mice. Front Biosci,2007,12(20):4722-4730.
[71]Hoetelmans R, Van Slooten HJ, Keijzer R, et al. Bcl-2 and Bax proteins are present in interphase nuclei of mammalian cells. Cell Death Differ,2000,7(4):384-392.
[72]Korsmeyer SJ, Shutter JR, Veis DJ, et al. Bcl-2/Bax:a rheostat that regulates an anti-oxidant pathway and cell death. Semin Cancer Biol,1993,4(6):327-332.
[73]Mathai JP, Germain M, Shore GC. BH3-only BIK regulates BAX, BAK-dependent release of Ca2+from endoplasmic reticulum stores and mitochondrial apoptosis during stress-induced cell death. J Biol Chem,2005,280(25):23829-23836.
[74]Chami M, Prandini A, Campanella M, et al. Bcl-2 and Bax exert opposing effects on Ca2+ signaling, which do not depend on their putative pore-forming region. J Biol Chem, 2004,279(52):54581-54589.
[75]Voehringer DW, Meyn RE. Redox aspects of Bcl-2 function. Antioxid Redox Signal, 2000,2(3):537-550.
[76]Metrailler-Ruchonnet I, Pagano A, Carnesecchi S, et al. Bcl-2 protects against hyperoxia-induced apoptosis through inhibition of the mitochondria-dependent pathway. Free Radic Biol Med,2007,42 (7):1062-1074.
[77]Ellerby LM, Ellerby HM, Park SM, et al. Shift of the cellular oxidation-reduction potential in neural cells expressing Bcl-2. J Neurochem,1996,67(3):1259-1267.
[78]Brooks C, Dong Z. Regulation of mitochondrial morphological dynamics during apoptosis by Bcl-2 family proteins:a key in Bak?. Cell Cycle,2007,6(24):3043-3047.
[79]樊丽琳,陈东风,胡辂,等.bcl-2、bax在大鼠非酒精性脂肪性肝病中的表达及意义.重庆医学,2007,36(8)：692-694.
[80]Vaux DL, Cory S, Adams JM. Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pro-B cells. Nature,1998,335 (6189):440-442.
[81]Sato H, Ozawa K, Iwata S, et al. Role of bcl-2 mRNA in homeostatic proliferation in circulating T-cells in human liver transplant patients after T-cell depletion. J Surg Res, 2005,127(2):123-130.
[82]Bermudez Y, Erasso D, Johnson NC, et al. Telomerase confers resistance to caspase-mediated apoptosis. Clin Interv Aging,2006,1(2):155-167.
[83]Malhi H, Bronk SF, Werneburg NW, et al. Free fatty acids induce JNK-dependent hepatocyte lipoapoptosis. J Biol Chem,2006,281(17):12093-12101.
[84]Fletcher JI, Huang DC. Controlling the cell death mediators Bax and Bak:puzzles and conundrums. Cell Cycle,2008,7(1):39-44.
[85]Debatin KM. Chronic lymphocytic leukemia:keeping cell death at bay. Cell,2007, 129(5):853-855.
[86]Wong CK, Chen Z, So KL, et al. Polycomb group protein RING IB is a direct substrate of Caspases-3 and-9. Biochim Biophys Acta,2007,1773(6):844-852.
[87]Bao Q, Lu W, Rabinowitz JD, et al. Calcium blocks formation of apoptosome by preventing nucleotide exchange in Apaf-1. Mol Cell,2007,25(2):181-192.
[88]张静,赵景民,周光德,等.非酒精性脂肪性肝炎肝细胞凋亡及凋亡相关蛋白的表达.解放军医学杂志,2008,33(5)：521-523.
[89]Tsukahara S, Yamamoto S, Tin-Tin-Win-Shwe, et al. Inhalation of low-level formaldehyde increases the bcl-2/bax expression ratio in the hippocampus of immunologically sensitized mice. Neuroimmunomodulation,2006,13(2):63-68.
[90]殷果华,殷云勤,赵和平,等.降脂益肝冲剂治疗脂肪肝的实验观察.中华实验和临床病毒学杂志,2000,14(2)：193-194.
[91]胡义扬,刘平,刘成,等.丹参提取物对CC1-4和DMN诱导的大鼠肝纤维化的影响. 上海中医药杂志,1999(10)：7-10.
[92]洪学智.泽泻治疗非酒精性脂肪肝病药效及作用机理研究.[硕士学位论文].浙江大学,2006.
[93]李英,王晓素.脂肪肝的中医药治疗现状.甘肃中医,2006,19(3)：35-37.
[94]王德山,肖玉芳,董朝晖,等.枸杞抗实验性高血脂肝脂量效关系及毒性研究.辽宁中医杂志,1997,24(12)：567-568.
[95]李续娥,郭宝江.决明子蛋白质和葸醌苷对高脂血症大鼠血脂的影响.中国临床康复,2004,8(18)：3694-3695.
[96]李贵海,孙敬勇,张希林,等.山楂降血脂有效成分的实验研究.中草药,2002,33(1)：50-52.
[97]Bjorntorp P. Metabolic difference between visceral fat and subcutaneous abdominal fat. Diabetes Metab,2000,26(Suppl 3):10-12.
[1]Wieckowska A, Zein NN, Yerian LM, et al. In vivo assessment of liver cell apoptosis as a novel biomarker of disease severity in nonalcoholic fatty liver disease[J]. Hepatology, 2006,44(1):27-33.
[2]Tsukahara S, Yamamoto S, Tin-Tin-Win-Shwe, et al. Inhalation of low-level formaldehyde increases the Bcl-2/Bax expression ratio in the hippocampus of immunologically sensitized mice[J]. Neuroimmunomodulation,2006,13(2):63-68.
[3]Tsujimoto Y, Cossman J, Jaffe E, et al. Involvement of the bcl-2 gene in human follicular lymphoma[J]. Science,1985,228 (4706):1440-1443.
[4]Roset R, Ortet L, Gil-Gomez G. Role of Bcl-2 family members on apoptosis:what we have learned from knock-out mice [J]. Front Biosci,2007,12 (20):4722-4730.
[5]Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death[J]. Cell,1993,74 (4):609-619.
[6]Hoetelmans R, Van Slooten HJ, Keijzer R, et al. Bcl-2 and Bax proteins are present in interphasenuclei of mammalian cells[J]. Cell Death Differ,2000,7(4):384-392.
[7]陈东风,王春华.体外非酒精性脂肪肝细胞模型线粒体通透转换孔的变化及其与细胞凋亡的关系[J].中华肝脏病杂志,2007,15(11)：837-839.
[8]Dorscheid DR, Wojcik KR, Sun S, et al. Apoptosis of airway epithelial cells induced by corticosteroids[J]. Am J Respir Crit Care Med,2001,164(10):1939-1947.
[9]White SR, Dorscheid DR. Corticosteroid-induced apoptosis of airway epithelium:a potential mechanism for chronic airway epithelial damage in asthma[J]. Chest, 2002,122(6):278-284.
[10]Zamzami N, Kroemer G. Apopotosis:mitochondrial membrane permeabilization-the (w)hole story? [J]. Curr Biol,2003,13(2):71-73.
[11]Strasser A, O'Connor L, Dixit VM, et al. Apopotosis signaling[J]. Annu Rev Biochem, 2000,69 (1):217-245.
[12]Wei MC, Zong WX, Cheng EH, et al. Proapopptotic BAX and BAK:a requisite gateway to mitochondrial dysfunction and death[J]. Science,2001,292(5517):727-730.
[13]Boisvieux-Ulrich E, Sourdeval M, Marano F. CD437, a synthetic retinoid, induces apoptosis in human respiratory epithelial cells via caspase-independent mitochondrial and caspase-8-dependent pathways both up-regulated by JNK signaling pathway[J]. Exp Cell Res,2005,307(1):76-90.
[14]Rust C, Gores GJ. Apoptosis and liver disease[J]. Am J Med,2000,108(7):567-574.
[15]Hannun YA. Apoptosis and the dilemma of cancer chemotherapy[J]. Blood,1997, 89(6):1845-1853.
[16]Mathai JP, Germain M, Shore GC. BH3-only BIK regulates BAX, BAK-dependent release of Ca2+from endoplasmic reticulum stores and mitochondrial apoptosis during stress-induced cell death[J]. J Biol Chem,2005,280(25):23829-23836.
[17]Chami M, Prandini A, Campanella M, et al. Bcl-2 and Bax exert opposing effects on Ca2+ signaling, which do not depend on their putative pore-forming region[J]. J Biol Chem,2004,279(52):54581-54589.
[18]Voehringer DW, Meyn RE. Redox aspects of Bcl-2 function[J]. Antioxid Redox Signal, 2000,2(3):537-550.
[19]Metrailler-Ruchonnet I, Pagano A, Carnesecchi S, et al. Bcl-2 protects against hyperoxia-induced apoptosis through inhibition of the mitochondria-dependent pathway[J]. Free Radic BioIMed,2007,42 (7):1062-1074.
[20]Ellerby LM, Ellerby HM, Park SM, et al. Shift of the cellular oxidation-reduction potential in neural cells expressing Bcl-2[J]. J Neurochem,1996,67(3):1259-1267.
[21]Brooks C, Dong Z. Regulation of mitochondrial morphological dynamics during apoptosis by Bcl-2 family proteins:a key in Bak?[J]. Cell Cycle,2007,6 (24): 3043-3047.
[22]樊丽琳,陈东风,胡辂,等.bcl-2、bax在大鼠非酒精性脂肪性肝病中的表达及意义[J].重庆医学,2007,36(8)：692-694.
[23]Vaux DL, Cory S, Adams JM. Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pro-B cells[J]. Nature,1998,335 (6189): 440-442.
[24]Sato H, Ozawa K, Iwata S, et al. Role of bcl-2 mRNA in homeostatic proliferation in circulating T-cells in human liver transplant patients after T-cell depletion[J]. J Surg Res,2005,127(2):123-130.
[25]Bermudez Y, Erasso D, Johnson NC, et al. Telomerase confers resistance to caspase-mediated apoptosis[J]. Clin Interv Aging,2006,1(2):155-167.
[26]Malhi H, Bronk SF, Werneburg NW, et al. Free fatty acids induce JNK-dependent hepatocyte lipoapoptosis[J]. J Biol Chem,2006,281(17):12093-12101.
[27]Fletcher JI, Huang DC. Controlling the cell death mediators Bax and Bak:puzzles and conundrums[J]. Cell Cycle,2008,7(1):39-44.
[28]Debatin KM. Chronic lymphocytic leukemia:keeping cell death at bay[J]. Cell,2007, 129(5):853-855.
[29]Wong CK, Chen Z, So KL, et al. Polycomb group protein RING1B is a direct substrate of Caspases-3 and-9[J]. Biochim Biophys Acta,2007,1773(6):844-852.
[30]Bao Q, Lu W, Rabinowitz JD, et al. Calcium blocks formation of apoptosome by preventing nucleotide exchange in Apaf-1[J]. Mol Cell,2007,25(2):181-192.
[31]Rogerio F, Jordao H Jr, Vieira AS, et al. Bax and Bcl-2 expression and TUNEL labeling in lumbar enlargement of neonatal rats after sciatic axotomy and melatonin treatment[J]. Brain Res,2006,1112(1):80-90.
[32]张静,赵景民,周光德,等.非酒精性脂肪性肝炎肝细胞凋亡及凋亡相关蛋白的表达[J].解放军医学杂志,2008,33(5)：521-523.
[33]Higuchi M, Aggarwal BB, Yeh ET. Activation of CPP32-like protease in tumor necrosis factor-induced apoptosis is dependent on mitochondrial function[J]. J Clin Invest,1997,99 (7):1751-1758.