解毒化瘀中药对急性淋巴细胞白血病模型小鼠的改善作用
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摘要
背景:研究表明,白血病患者需要及早行化疗延缓病情进展,中草药对该病治疗效果甚微,还有研究持相反意见,认为中草药联合化疗可以一定程度提高患者的治愈率。由于中草药成分复杂,治疗的机制尚不明确。目的:探究解毒化瘀中药对急性淋巴细胞白血病模型小鼠的影响。方法:将白血病细胞株L1210和白血病耐药细胞分别从小鼠尾静脉注入体内制备急性淋巴细胞白血病模型,随机分为白血病模型组、解毒化瘀中药低浓度组、解毒化瘀中药高浓度组,另设正常对照组。造模成功后第2天按照10,50 mg/kg给药,连续7 d,于每周一早晨7点取尾静脉血,连续5周,采用MTT检测白血病耐药细胞生存率,分析血细胞种类及血象;Westernblot法检测小鼠骨髓中NF-κB/P65、NF-κB/P50、IκBα、PTEN、Akt、Bcl-2及p-Akt蛋白的表达。结果与结论:①与白血病模型组比较,解毒化瘀中药低、高浓度组白血病耐药细胞存活率显著降低,差异有显著性意义(P <0.01);②与模型组比较,解毒化瘀中药低、高浓度组小鼠骨髓中NF-κB/P65、NF-κB/P50、IκBα、Bcl-2、Akt、p-Akt蛋白表达量显著降低,PTEN蛋白表达量显著升高,差异有显著性意义(P <0.01);③与模型组相比,解毒化瘀中药低、高浓度组白细胞、淋巴细胞、L1210细胞显著降低,中性粒细胞显著升高,差异有显著性意义(P<0.01);④结果表明,解毒化瘀中药通过阻断PI3K/Akt及NF-κB信号通路实现抑制L1210细胞增殖效应,从而缓解和治疗急性淋巴细胞白血病。
BACKGROUND: Studies have shown that early chemotherapy is essential for delaying the progression of leukemia. Chinese herbal medicine has little effect on leukemia, but some studies hold the opposite opinion that Chinese herbal medicine combined with chemotherapy can improve the cure rate of patients to a certain extent. Due to the complex composition, the therapeutic mechanism of Chinese herbal medicines is still unclear.OBJECTIVE: To explore the effects of Chinese medicine for detoxifying and removing blood stasis in mice with acute lymphoblastic leukemia.METHODS: Leukemia cell line L1210 and leukemia drug-resistant cells were injected into mice via the tail vein to prepare mouse models of acute lymphoblastic leukemia. These models were then divided into leukemia model group, low-and high-concentration Chinese medicine for detoxifying and removing blood stasis groups, and normal group was set for controls. At the second day after modeling, Chinese medicine for detoxifying and removing blood stasis(10 and 50 mg/kg) was given for 7 days in the low-and high-concentration treatment group, respectively.Blood samples from the tail vein were taken at 7:00 a.m. on Monday, for 5 consecutive weeks. MTT was used to detect the survival rate of leukemia drug-resistant cells, and blood cell types and hemogram indexes were analyzed. The expression levels of NF-κB/P65, NF-κB/P50,IκBα, phosphatase and tensin homolog deleted on chromosome ten(PTEN), Akt, Bcl-2 and p-Akt were detected by western blot assay.RESULTS AND CONCLUSION:(1) Compared with the leukemia model group, the survival rate of leukemia drug-resistant cells was significantly decreased in the low-and high-concentration treatment groups.(2) Compared with the leukemia model group, the expression levels of NF-κB/P65, NF-κB/P50, IκBα, Bcl-2, Akt and p-Akt were significantly decreased in the low-and high-concentration treatment groups,while the expression level of PTEN increased significantly(P < 0.01).(3) Compared with the leukemia model group, the leukocytes, the number of white blood cells, lymphocytes and L1210 cells in the low-and high-concentration treatment groups was significantly decreased,and the number of neutrophils was significantly increased(P < 0.01).(4) To conclude, Chinese medicine for detoxifying and removing blood stasis can inhibit the proliferation of L1210 cells by blocking the PI3 K/Akt and NF-κB signaling pathway to alleviate and treat acute lymphoblastic leukemia.
BACKGROUND: Studies have shown that early chemotherapy is essential for delaying the progression of leukemia. Chinese herbal medicine has little effect on leukemia, but some studies hold the opposite opinion that Chinese herbal medicine combined with chemotherapy can improve the cure rate of patients to a certain extent. Due to the complex composition, the therapeutic mechanism of Chinese herbal medicines is still unclear.OBJECTIVE: To explore the effects of Chinese medicine for detoxifying and removing blood stasis in mice with acute lymphoblastic leukemia.METHODS: Leukemia cell line L1210 and leukemia drug-resistant cells were injected into mice via the tail vein to prepare mouse models of acute lymphoblastic leukemia. These models were then divided into leukemia model group, low-and high-concentration Chinese medicine for detoxifying and removing blood stasis groups, and normal group was set for controls. At the second day after modeling, Chinese medicine for detoxifying and removing blood stasis(10 and 50 mg/kg) was given for 7 days in the low-and high-concentration treatment group, respectively.Blood samples from the tail vein were taken at 7:00 a.m. on Monday, for 5 consecutive weeks. MTT was used to detect the survival rate of leukemia drug-resistant cells, and blood cell types and hemogram indexes were analyzed. The expression levels of NF-κB/P65, NF-κB/P50,IκBα, phosphatase and tensin homolog deleted on chromosome ten(PTEN), Akt, Bcl-2 and p-Akt were detected by western blot assay.RESULTS AND CONCLUSION:(1) Compared with the leukemia model group, the survival rate of leukemia drug-resistant cells was significantly decreased in the low-and high-concentration treatment groups.(2) Compared with the leukemia model group, the expression levels of NF-κB/P65, NF-κB/P50, IκBα, Bcl-2, Akt and p-Akt were significantly decreased in the low-and high-concentration treatment groups,while the expression level of PTEN increased significantly(P < 0.01).(3) Compared with the leukemia model group, the leukocytes, the number of white blood cells, lymphocytes and L1210 cells in the low-and high-concentration treatment groups was significantly decreased,and the number of neutrophils was significantly increased(P < 0.01).(4) To conclude, Chinese medicine for detoxifying and removing blood stasis can inhibit the proliferation of L1210 cells by blocking the PI3 K/Akt and NF-κB signaling pathway to alleviate and treat acute lymphoblastic leukemia.
引文
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[11]杨小娜,郭建美,姚海英,等.中医药治疗白血病的研究进展[J].现代中西医结合杂志,2016,25(10):1134-1137.
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[13]周建军,乐秀芳.评价抗癌物质活性的改良MTT方法[J].中国医药工业杂志,1993(10):455-457.
[14]马武开,姚血明,唐芳,等.解毒化瘀药调控白血病HL60/adr细胞NF-κB-survivin通路研究[J].北京中医药大学学报,2012,35(1):25-28.
[15]Chang HY,Pan KL,Ma FC,et al.The study on reversing mechanism of multidrug resistance of K562/A02 cell line by curcumin and erythromycin.Zhonghua Xue Ye Xue Za Zhi.2006;27(4):254-258.
[16]蒋卉男,刘卓刚,胡荣.Embelin逆转K562/D细胞对柔红霉素耐药与P-gp及MDR1 mRNA无关[J].中国实验血液学杂志,2017,25(5):1342-1349.
[17]Li X,Wu C,Chen N,et al.PI3K/Akt/mTOR signaling pathway and targeted therapy for glioblastoma.Oncotarget.2016;7(22):33440-33450.
[18]Zhou ZQ,Li YL,Ao ZB,et al.Baicalin protects neonatal rat brains against hypoxic-ischemic injury by upregulating glutamate transporter 1 via the phosphoinositide3-kinase/protein kinase B signaling pathway.Neural Regen Res.2017;12(10):1625-1631.
[19]Yan LX,Liu YH,Xiang JW,et al.PIK3R1 targeting by miR-21suppresses tumor cell migration and invasion by reducing PI3K/AKT signaling and reversing EMT,and predicts clinical outcome of breast cancer.Int J Oncol.2016;48(2):471-484.
[20]严匡华,陈龙,严风梦.下调miR-21抑制PI3K/AKT信号通路对白血病细胞增殖凋亡的影响[J].中国免疫学杂志,2018,34(3):441-445.
[21]白炎亮,张茵,于润红,等.PI3K/Akt传通路在急性白血病细胞中的表达及其意义[J].中国实用医刊,2017,44(19):75-79.
[22]Canté-Barrett K,Spijkers-Hagelstein JA,Buijs-Gladdines JG,et al.MEK and PI3K-AKT inhibitors synergistically block activated IL7 receptor signaling in T-cell acute lymphoblastic leukemia.Leukemia.2016;30(9):1832-1843.
[23]师丽晓,王建华,师晓东.PI3k/Akt/mTOR信号通路与儿童急性T淋巴细胞白血病[J].中国实验血液学杂志,2016,24(4):1269-1274.
[24]饶佳,卢玮,王芸芸,等.bcl-2基因过表达致急性髓系白血病对柔红霉素耐药机制的初步探讨[J].安徽医科大学学报,2017,52(3):369-372.
[25]李攀登,王永红,肖芸,等.氢醌对人白血病细胞凋亡及相关蛋白Bcl-2、Bax及Caspase-3表达的影响[J].现代预防医学,2018,45(5):878-882.
[26]杨君义,徐飞.治疗慢性淋巴细胞白血病新药--BCL-2抑制剂venetoclax[J].中国新药与临床杂志,2017,8(3):131-133.
[2]Kansagra A,Litzow M.Treatment of Young Adults with Acute Lymphoblastic Leukemia.Curr Hematol Malig Rep.2017;12(3):187-196.
[3]Chen XP,Li W,Xiao XF,et al.Phytochemical and pharmacological studies on Radix Angelica sinensis.Chin JNat Med.2013;11(6):577-587.
[4]Ma H,Cheng L,Hao K,et al.Reversal effect of ST6GAL 1 on multidrug resistance in human leukemia by regulating the PI3K/Akt pathway and the expression of P-gp and MRP1.PLoS One.2014;9(1):e85113.
[5]Sun Y,Peng R,Peng H,et al.miR-451 suppresses the NF-kappaB-mediated proinflammatory molecules expression through inhibiting LMP7 in diabetic nephropathy.Mol Cell Endocrinol.2016;433:75-86.
[6]Sun BT,Zheng LH,Bao YL,et al.Reversal effect of Dioscin on multidrug resistance in human hepatoma HepG2/adriamycin cells.Eur J Pharmacol.2011;654(2):129-134.
[7]Hien TT,Kim HG,Han EH,et al.Molecular mechanism of suppression of MDR1 by puerarin from Pueraria lobata via NF-kappaB pathway and cAMP-responsive element transcriptional activity-dependent up-regulation of AMP-activated protein kinase in breast cancer MCF-7/adr cells.Mol Nutr Food Res.2010;54(7):918-928.
[8]张献,冯欣.PI3K/AKT/mTOR信号通路抑制剂在宫颈癌治疗中的研究进展[J].现代药物与临床,2018,33(5):1278-1284.
[9]吴宥熹,徐俊,刘立亚,等.PI3K-Akt-mTOR信号通路介导的细胞自噬在荭草苷抗小鼠心肌缺血再灌注损伤中的作用研究[J].时珍国医国药,2017,21(1):20-24.
[10]赖芸,陈子珺.有毒中药抗白血病的作用及机制研究进展[J].中成药,2016,38(4):875-879.
[11]杨小娜,郭建美,姚海英,等.中医药治疗白血病的研究进展[J].现代中西医结合杂志,2016,25(10):1134-1137.
[12]宋艳秋,刘敏,李薇,等.K562/NOD-SCID小鼠白血病模型的建立[J].中国实验血液学杂志,2007,15(1):16-19.
[13]周建军,乐秀芳.评价抗癌物质活性的改良MTT方法[J].中国医药工业杂志,1993(10):455-457.
[14]马武开,姚血明,唐芳,等.解毒化瘀药调控白血病HL60/adr细胞NF-κB-survivin通路研究[J].北京中医药大学学报,2012,35(1):25-28.
[15]Chang HY,Pan KL,Ma FC,et al.The study on reversing mechanism of multidrug resistance of K562/A02 cell line by curcumin and erythromycin.Zhonghua Xue Ye Xue Za Zhi.2006;27(4):254-258.
[16]蒋卉男,刘卓刚,胡荣.Embelin逆转K562/D细胞对柔红霉素耐药与P-gp及MDR1 mRNA无关[J].中国实验血液学杂志,2017,25(5):1342-1349.
[17]Li X,Wu C,Chen N,et al.PI3K/Akt/mTOR signaling pathway and targeted therapy for glioblastoma.Oncotarget.2016;7(22):33440-33450.
[18]Zhou ZQ,Li YL,Ao ZB,et al.Baicalin protects neonatal rat brains against hypoxic-ischemic injury by upregulating glutamate transporter 1 via the phosphoinositide3-kinase/protein kinase B signaling pathway.Neural Regen Res.2017;12(10):1625-1631.
[19]Yan LX,Liu YH,Xiang JW,et al.PIK3R1 targeting by miR-21suppresses tumor cell migration and invasion by reducing PI3K/AKT signaling and reversing EMT,and predicts clinical outcome of breast cancer.Int J Oncol.2016;48(2):471-484.
[20]严匡华,陈龙,严风梦.下调miR-21抑制PI3K/AKT信号通路对白血病细胞增殖凋亡的影响[J].中国免疫学杂志,2018,34(3):441-445.
[21]白炎亮,张茵,于润红,等.PI3K/Akt传通路在急性白血病细胞中的表达及其意义[J].中国实用医刊,2017,44(19):75-79.
[22]Canté-Barrett K,Spijkers-Hagelstein JA,Buijs-Gladdines JG,et al.MEK and PI3K-AKT inhibitors synergistically block activated IL7 receptor signaling in T-cell acute lymphoblastic leukemia.Leukemia.2016;30(9):1832-1843.
[23]师丽晓,王建华,师晓东.PI3k/Akt/mTOR信号通路与儿童急性T淋巴细胞白血病[J].中国实验血液学杂志,2016,24(4):1269-1274.
[24]饶佳,卢玮,王芸芸,等.bcl-2基因过表达致急性髓系白血病对柔红霉素耐药机制的初步探讨[J].安徽医科大学学报,2017,52(3):369-372.
[25]李攀登,王永红,肖芸,等.氢醌对人白血病细胞凋亡及相关蛋白Bcl-2、Bax及Caspase-3表达的影响[J].现代预防医学,2018,45(5):878-882.
[26]杨君义,徐飞.治疗慢性淋巴细胞白血病新药--BCL-2抑制剂venetoclax[J].中国新药与临床杂志,2017,8(3):131-133.