藏族药“亚吉玛”(裸茎金腰)提取物对ANIT致肝内胆汁淤积小鼠的保护作用及机制的探讨
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摘要
该文采用α-萘异硫氰酸酯致小鼠肝内胆汁淤积模型,对藏族医临床泻胆热症常用药物"亚吉玛"(裸茎金腰Chrysosplenium nudicaule)提取物(ECN)对小鼠肝损伤的保护作用及其作用机制进行了探讨。结果表明,ECN能剂量依赖性地降低模型小鼠血清中AST,ALT,ALP,DBIL,TBIL,TBA的含量水平,升高小鼠肝组织匀浆SOD和GSH-Px的活性(P<0. 01,P<0. 05),同时降低MDA的含量(P<0. 01,P<0. 05)。小鼠肝脏病理切片显示,ECN可改善模型小鼠肝组织病理变化。mRNA分析结果表明,ECN能显著改变胆汁酸代谢相关基因的表达水平,如BSEP,FXR,MRP2 mRNA表达显著上调(P<0. 01,P<0. 05),同时CYP7A1的表达显著下调(P<0. 01,P<0. 05)。上述结果证实了ECN对小鼠胆汁淤积型肝损伤的保护作用,并提示其作用机制可能与激活FXR及其靶基因,减少胆汁酸的合成,增加胆汁酸的排泄有关。该研究为特色藏族药亚吉玛在藏族医药中的临床应用提供了现代药理学依据。
Chrysosplenium nudicaule,Tibetan name " Yajima",is recorded as an effective medicine for the treatment of liver and gallbladder diseases by Tibetan Pharmacopoeia published in the past dynasties,but its traditional efficacy has not yet been investigated by means of modern pharmacological research methods. In this paper,the protective effect of extract of C. nudicaule(ECN) on liver injury in mice was observed by using the mice model of intrahepatic cholestasis(IC) induced by α-naphthyl isothiocyanate(ANIT) and the possible mechanism by which ECN work as the therapeutic agent was discussed. The results showed that the serum levels of AST,ALT,ALP,DBIL,TBIL and TBA of the model mice were notably reduced in dose-dependent manner(P<0. 01,P<0. 05). The activity of SOD and GSH-Px in the liver homogenate of mice was increased,while the content of MDA was decreased(P<0. 01,P<0. 05).Pathological examination of liver in mice showed that ECN could improve the pathological changes of liver tissue in mice. The mRNA expression level of genes related to bile acid metabolism were detected by RT-PCR and the results suggested that ECN could significantly increase the expression of genes such as BSEP,FXR and MRP2(P<0. 01,P<0. 05),meanwhile significantly reduce the expression of CYP7 A1(P<0. 01,P<0. 05). These results confirmed the protective effect of ECN on intrahepatic cholestasis-induced liver injury in mice,and indicated that the mechanism may be related to activating FXR and its target genes,reducing bile acid synthesis and increasing bile acid excretion. This study provides a modern pharmacological basis for the clinical application of Yajima in Tibetan medicine.
Chrysosplenium nudicaule,Tibetan name " Yajima",is recorded as an effective medicine for the treatment of liver and gallbladder diseases by Tibetan Pharmacopoeia published in the past dynasties,but its traditional efficacy has not yet been investigated by means of modern pharmacological research methods. In this paper,the protective effect of extract of C. nudicaule(ECN) on liver injury in mice was observed by using the mice model of intrahepatic cholestasis(IC) induced by α-naphthyl isothiocyanate(ANIT) and the possible mechanism by which ECN work as the therapeutic agent was discussed. The results showed that the serum levels of AST,ALT,ALP,DBIL,TBIL and TBA of the model mice were notably reduced in dose-dependent manner(P<0. 01,P<0. 05). The activity of SOD and GSH-Px in the liver homogenate of mice was increased,while the content of MDA was decreased(P<0. 01,P<0. 05).Pathological examination of liver in mice showed that ECN could improve the pathological changes of liver tissue in mice. The mRNA expression level of genes related to bile acid metabolism were detected by RT-PCR and the results suggested that ECN could significantly increase the expression of genes such as BSEP,FXR and MRP2(P<0. 01,P<0. 05),meanwhile significantly reduce the expression of CYP7 A1(P<0. 01,P<0. 05). These results confirmed the protective effect of ECN on intrahepatic cholestasis-induced liver injury in mice,and indicated that the mechanism may be related to activating FXR and its target genes,reducing bile acid synthesis and increasing bile acid excretion. This study provides a modern pharmacological basis for the clinical application of Yajima in Tibetan medicine.
引文
[1]胆汁淤积性肝病诊断治疗专家共识2015年更新专家委员会.胆汁淤积性肝病诊断治疗专家共识:2015年更新[J].临床肝胆病杂志,2015,10(31):1563.
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[3]韩平,田德安.肝内胆汁淤积发病机制及药物治疗新进展[J].胃肠病学和肝病学杂志,2016,25(5):584.
[4] Adcock I M,Mumby S. Glucocorticoids[J]. Handb Exp Pharmacol,2017,237:171.
[5] Geraets S E,Gosens T. The intra-articular glucocorticoid injection:short-term success with potential side effects[J]. Ned Tijdschr Geneeskd,2016,160:814.
[6]平键,陈红云,周扬,等.骨髓间充质干细胞移植对四氯化碳大鼠肝损伤的影响及一贯煎的干预作用[J].中国中药杂志,2018,43(19):3905.
[7]李姗姗,李博,张寿文,等.藏药蒂达的6种主要基原植物提取物对急性肝内胆汁淤积小鼠药理作用的比较研究[J].中药新药与临床药理,2017,28(3):314.
[8]饶雅琪,张丽,曾金祥,等.长毛风毛菊醇提物对ANIT所致小鼠肝损伤的保肝作用研究[J].中国新药杂志,2017,26(13):1566.
[9]蒂玛尔·丹增彭措.晶珠本草[M].上海:上海科学技术出版社,1986.
[10]杨永昌.藏药志[M].西宁:青海人民出版社,1991.
[11]中华人民共和国卫生部药典委员会.部颁药品标准·藏药分册.第1册[S].北京:中国医药科技出版社,1995:61.
[12]西藏、青海、四川、甘肃、云南、新疆卫生局.藏药标准.第一、二分册合编本[S].西宁:青海人民出版社,1992:55.
[13] Tanaka Y,Aleksunes L M,Cui Y J,et al. ANIT-induced intrahepatic cholestasis alters hepatobiliary transporter expression via Nrf2-dependent and independent signaling[J]. Toxicol Sci,2009,108(2):247.
[14]解静.赶黄草提取物对胆汁淤积性肝细胞损伤的保护作用[D].泸州:西南医科大学,2016.
[15] Dreher D,Junod A F. Differential effects of superoxide,hydrogen peroxide,and hydroxyl radical on intracellular calcium in human endothelial cells[J]. J Cell Physiol,1995,162:147.
[16] Hou J,You G,Xu Y,et al. Antioxidant enzyme activities as biomarkers of fluvial biofilm to Zn O NPs ecotoxicity and the Integrated Biomarker Responses(IBR)assessment[J]. Ecotoxicol Environ Saf,2016,133:10.
[17]王凯,眭丹娟,王长锁,等. 5种石斛对四氯化碳致小鼠急性肝损伤的保护作用[J].中国中药杂志, 2017, 42(10):1945.
[18]尹萍,崔鹤蓉,章从恩,等.六味五灵片对小鼠急性乙醇性肝损伤的保护作用及机制初步研究[J].中国中药杂志,2016,41(19):3637.
[19]龙爽,刘绍勇,徐英.痰热清注射液及5种中间体对四氯化碳致急性肝损伤大鼠的保护作用探讨[J].中国实验方剂学杂志,2018,24(11):73.
[20] Lee E A,Lee D I,Kim H Y,et al. Cyp7a1 is continuously increased with disrupted Fxr-mediated feedback inhibition in hypercholesterolemic TALLYHO/Jng mice[J]. Biochim Biophys Acta,2018,1863(1):20.
[21] Liu B,Li Y,Ji H,et al. Glutamine attenuates obstructive cholestasis in rats via farnesoid X receptor-mediated[J]. Can J Physiol Pharmacol,2017,95(2):215.
[22] Wang L L,Wu G,Wu F H,et al. Geniposide attenuates ANITinduced cholestasis through regulation of transporters and enzymes involved in bile acids homeostasis in rats[J]. J Ethnopharmacol,2017,196:178.
[23] Ding L,Zhang B,Li J,et al. Beneficial effect of resveratrol onα-naphthyl isothiocyanate-induced cholestasis via regulation of the FXR pathway[J]. Mol Med Rep,2018,17(1):1863.
[24] Yang F,Tang X,Ding L,et al. Curcumin protects ANITinduced cholestasis through signaling pathway of FXR-regulated bile acid and inflammation[J]. Sci Rep,2016,6:33052.
[2]樊雪强,王彩生,刘金涛,等.熊去氧胆酸在原发性胆汁性肝硬化临床治疗中的进展[J].中国医药导报,2016,13(24):58.
[3]韩平,田德安.肝内胆汁淤积发病机制及药物治疗新进展[J].胃肠病学和肝病学杂志,2016,25(5):584.
[4] Adcock I M,Mumby S. Glucocorticoids[J]. Handb Exp Pharmacol,2017,237:171.
[5] Geraets S E,Gosens T. The intra-articular glucocorticoid injection:short-term success with potential side effects[J]. Ned Tijdschr Geneeskd,2016,160:814.
[6]平键,陈红云,周扬,等.骨髓间充质干细胞移植对四氯化碳大鼠肝损伤的影响及一贯煎的干预作用[J].中国中药杂志,2018,43(19):3905.
[7]李姗姗,李博,张寿文,等.藏药蒂达的6种主要基原植物提取物对急性肝内胆汁淤积小鼠药理作用的比较研究[J].中药新药与临床药理,2017,28(3):314.
[8]饶雅琪,张丽,曾金祥,等.长毛风毛菊醇提物对ANIT所致小鼠肝损伤的保肝作用研究[J].中国新药杂志,2017,26(13):1566.
[9]蒂玛尔·丹增彭措.晶珠本草[M].上海:上海科学技术出版社,1986.
[10]杨永昌.藏药志[M].西宁:青海人民出版社,1991.
[11]中华人民共和国卫生部药典委员会.部颁药品标准·藏药分册.第1册[S].北京:中国医药科技出版社,1995:61.
[12]西藏、青海、四川、甘肃、云南、新疆卫生局.藏药标准.第一、二分册合编本[S].西宁:青海人民出版社,1992:55.
[13] Tanaka Y,Aleksunes L M,Cui Y J,et al. ANIT-induced intrahepatic cholestasis alters hepatobiliary transporter expression via Nrf2-dependent and independent signaling[J]. Toxicol Sci,2009,108(2):247.
[14]解静.赶黄草提取物对胆汁淤积性肝细胞损伤的保护作用[D].泸州:西南医科大学,2016.
[15] Dreher D,Junod A F. Differential effects of superoxide,hydrogen peroxide,and hydroxyl radical on intracellular calcium in human endothelial cells[J]. J Cell Physiol,1995,162:147.
[16] Hou J,You G,Xu Y,et al. Antioxidant enzyme activities as biomarkers of fluvial biofilm to Zn O NPs ecotoxicity and the Integrated Biomarker Responses(IBR)assessment[J]. Ecotoxicol Environ Saf,2016,133:10.
[17]王凯,眭丹娟,王长锁,等. 5种石斛对四氯化碳致小鼠急性肝损伤的保护作用[J].中国中药杂志, 2017, 42(10):1945.
[18]尹萍,崔鹤蓉,章从恩,等.六味五灵片对小鼠急性乙醇性肝损伤的保护作用及机制初步研究[J].中国中药杂志,2016,41(19):3637.
[19]龙爽,刘绍勇,徐英.痰热清注射液及5种中间体对四氯化碳致急性肝损伤大鼠的保护作用探讨[J].中国实验方剂学杂志,2018,24(11):73.
[20] Lee E A,Lee D I,Kim H Y,et al. Cyp7a1 is continuously increased with disrupted Fxr-mediated feedback inhibition in hypercholesterolemic TALLYHO/Jng mice[J]. Biochim Biophys Acta,2018,1863(1):20.
[21] Liu B,Li Y,Ji H,et al. Glutamine attenuates obstructive cholestasis in rats via farnesoid X receptor-mediated[J]. Can J Physiol Pharmacol,2017,95(2):215.
[22] Wang L L,Wu G,Wu F H,et al. Geniposide attenuates ANITinduced cholestasis through regulation of transporters and enzymes involved in bile acids homeostasis in rats[J]. J Ethnopharmacol,2017,196:178.
[23] Ding L,Zhang B,Li J,et al. Beneficial effect of resveratrol onα-naphthyl isothiocyanate-induced cholestasis via regulation of the FXR pathway[J]. Mol Med Rep,2018,17(1):1863.
[24] Yang F,Tang X,Ding L,et al. Curcumin protects ANITinduced cholestasis through signaling pathway of FXR-regulated bile acid and inflammation[J]. Sci Rep,2016,6:33052.