肝宁方含药血清对毒胡萝卜素诱导大鼠肝细胞内质网应激的影响Effect of medicated serum of Ganning Decoction on hepatocyte endoplasmic reticulum stress of rats innduced by thapsigargin
朱朝阳;王建超;银艳桃;文彬;邓鑫;
摘要(Abstract):
目的研究肝宁方含药血清对毒胡萝卜素诱导的大鼠肝细胞BRL-3A内质网应激的影响及其相关机制。方法将30只SD大鼠平分为3组,其中10只予肝宁方25 g/(kg·d)灌胃,另2组大鼠以等量生理盐水灌胃,均2次/d,连续灌胃10 d后取血离心,获得含肝宁方大鼠血清及正常大鼠血清。取大鼠肝细胞BRL-3A细胞株,分为正常组、模型组、肝宁方组。正常组加入正常大鼠血清培养,模型组加入1μmol/L毒胡萝卜素及正常大鼠血清培养,肝宁方组加入1μmol/L毒胡萝卜素及含肝宁方大鼠血清培养。CCK8法检测BRL-3A细胞存活率,Fura-3/AM钙离子荧光探针测定BRL-3A细胞胞浆内Ca~(2+)水平,谷草转氨酶(ALT)、谷丙转氨酶(AST)试剂盒检测BRL-3A细胞中ALT、AST水平;RT-PCR法检测BRL-3A细胞内质网应激相关基因GRP78、PERK、CHOP、Bcl-2 mRNA表达水平。结果与正常组比较,模型组、肝宁方组BRL-3A细胞存活率及Bcl-2 mRNA表达水平均明显降低(P均<0.05),细胞胞浆内Ca~(2+)水平、细胞中ALT和AST水平及GRP78、PERK、CHOP mRNA表达水平均显著升高(P均<0.05);肝宁方组RL-3A细胞存活率及Bcl-2 mRNA表达水平均显著高于模型组(P均<0.05),细胞胞浆内Ca~(2+)水平、细胞中ALT和AST水平及GRP78 mRNA、PERK mRNA、CHOP mRNA表达水平均明显低于模型组(P均<0.05)。结论肝宁方可减弱内质网的Ca~(2+)流失,抑制内质网应激相关的GRP78、PERK、CHOP mRNA表达,上调Bcl-2mRNA表达,保护BRL-3A细胞并提高其存活率。
关键词(KeyWords): 肝宁方;大鼠肝细胞;内质网应激
基金项目(Foundation): 广西医疗卫生适宜技术开发与推广应用课题(s201503);; 国家自然科学基金资助项目(81360532)
作者(Author): 朱朝阳;王建超;银艳桃;文彬;邓鑫;
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参考文献(References):
- [1] Bravo R,Parra V,Gatica D,et al.Endoplasmic reticulum and the unfolded protein response:dynamics and metabolic integration[J].Int Rev Cell Mol Biol,2013,301(10):215-290.
- [2] Malhi H,Kaufman R J.Endoplasmic reticulum stress in liver disease[J].J Hepatol,2011,54(4):795-809.
- [3] Deng X,Liang J,Wu F S,et al.Effects of the Ganning Formula on Liver Fibrosis in Patients With Chronic Hepatitis B[J].J Trad Chin Med,2011,31(4):282-287.
- [4] 陈小聪.肝宁方对慢性乙型肝炎患者细胞免疫功能的影响[D].南宁:广西中医药大学,2017.
- [5] Guo B,Zhang W,Tume R K,et al.Disorder of endoplasmic reticulum calcium channel components is associated with the increased apoptotic potential in pale,soft,exudative pork[J].Meat Sci,2016,115:34-40.
- [6] Pagliassotti,Michael J.Endoplasmic Reticulum Stress in Nonalcoholic Fatty Liver Disease[J].Annu Rev Nutr,2012,32(1):17-33.
- [7] Lee A S.The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress[J].Methods (Amsterdam),2005,35(4):380-381.
- [8] Ji Y,Zhao Z,Cai T,et al.Liraglutide alleviates diabetic cardiomyopathy by blocking CHOP triggered apoptosis via the inhibition of the IRE-α pathway[J].Mol Med Rep,2014,9(4):1254-1258.
- [9] Ashraf N U,Sheikh T A.Endoplasmic reticulum stress and Oxidative stress in the pathogenesis of Non-alcoholic fatty liver disease[J].Free Radic Res,2015,49(12):1405-1418.
- [10] Lebeaupin C,Vallée D,Hazari Y,et al.Endoplasmic reticulum stress signalling and the pathogenesis of non-alcoholic fatty liver disease[J].J Hepatol,2018,69(4):927-947.
- [11] Wang L,Chen J,Ning C,et al.Endoplasmic Reticulum Stress Related Molecular Mechanisms in Nonalcoholic Fatty Liver Disease (NAFLD)[J].Curr Drug Targets,2018,19(9):1087-1094.
- [12] Maiers J L,Malhi H.Endoplasmic Reticulum Stress in Metabolic Liver Diseases and Hepatic Fibrosis[J].Semin Liver Dis,2019 ,39(2):235-248.
- [13] Chan C C,Lee K C,Huang Y H,et al.Regulation by resveratrol of the cellular factors mediating liver damage and regeneration after acute toxic liver injury[J].J Gastroenterol Hepatol,2014,29(3):603-613.
- [14] Hardwick J M,Soane L.Multiple Functions of BCL-2 Family Proteins[J].Cold Spring Harb Perspect Biol,2013,5(2):a008722.
- [15] Canová N K,Kmoníková E,Martínek J,et al.Thapsigargin,a selective inhibitor of sarco-endoplasmic reticulum Ca2+-ATPases,modulates nitric oxide production and cell death of primary rat hepatocytes in culture[J].Cell Biol Toxicol,2007,23(5):337-354.
- [16] 王键,宋瑞鹏,孙丹.黄芪甲苷对大鼠肝星状细胞氧化损伤的作用研究[J].解放军医药杂志,2018,30(9):1-4.
- [17] 王珏,李琴,李颖.黄芪总皂苷对CCl-4诱导急性肝损伤小鼠保护作用及机制初探[J].中药药理与临床,2018,34(6):70-73.
- [18] 李艳,伊航,蔡轶伦,等.黄芪总黄酮对四氯化碳诱导大鼠肝纤维化的影响[J].中成药,2019,41(7):1710-1713.
- [19] 钱隆,刘洋,李冰心,等.白术多糖可能通过Toll样受体4信号通路缓解环磷酰胺诱导的雏鹅肝脏损伤[J].动物营养学报,2019,31(2):764-774.
- [20] 车财妍,李红山,应豪,等.白术多糖对非酒精性脂肪性肝炎的防治作用研究[J].中华中医药学刊,2017,35(7):1801-1803.
- [21] 任艳青,牛丽颖,田宇柔,等.白芍配方颗粒对四氯化碳诱导的LO2细胞损伤的影响[J].解放军医学杂志,2016,41(5):378-383.
- [22] 林柳兵,阙任烨,刘进锴,等.柴胡皂苷d对活化的HSC-T6细胞MMP-1、TIMP-1表达的影响及其分子机制[J].世界华人消化杂志,2016,24(8):1159-1165.
- [23] 黄小强,张雪,熊丽樱,等.泽泻多糖对四氯化碳致小鼠急性肝损伤的保护作用[J].时珍国医国药,2017,28(6):1300-1302.
- [24] 宋之臻.当归多糖肝靶向作用及保护急性肝损伤机理的研究[D].武汉:华中科技大学,2016.
- [25] 熊莎,高建蓉,胡祖良,等.鳖甲提取物对抑制TGF-β诱导的大鼠肝星状细胞活化的影响[J].中国实验方剂学杂志,2017,23(19):155-159.
- [26] 吴军,尚瑞.茵陈水提物对牛血清白蛋白诱导的肝损伤大鼠的保护作用研究[J].实用肝脏病杂志,2017,20(1):51-54.
- [27] 唐爱存,陈兆霓,卢秋玉,等.葫芦茶苷对肝损伤大鼠肝组织Caspase-3与Caspase-8活性的影响及保肝作用研究[J].中华中医药学刊,2017,35(3):689-692.
- [28] 陈少锋,黄琳芸,钟鸣,等.瑶药“猛老虎”对大鼠免疫性肝纤维化影响的实验研究[J].时珍国医国药,2008,19(8):1813-1815.
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