姜黄素协同二甲双胍抑制肝星状细胞LX-2自噬及促进凋亡的研究
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摘要
目的探讨姜黄素协同二甲双胍对肝星状细胞LX-2自噬活性、凋亡的影响。方法将人肝星状细胞LX-2经DMEM培养基培养后分为4组:空白对照组、阴性对照组、姜黄素处理组(10μM)、姜黄素联合二甲双胍组:姜黄素(10μM)+二甲双胍(10mM),48h后通过Western blot法检测各组细胞自噬相关蛋白(LC3B、p62),MTS检测方法检测细胞增殖,流式细胞仪检测LX-2凋亡。结果与空白对照组、阴性对照组相比,姜黄素处理组可抑制LX-2自噬活性并促进其凋亡,且姜黄素协同二甲双胍组效应更强,差异均具有统计学意义(P<0.05)。结论姜黄素协同二甲双胍能抑制肝星状细胞LX-2自噬活性并促进其凋亡。
Objective To investigate the effect of curcumin and metformin on the autophagy and apoptosis of hepatic stellate cells(LX-2). Methods Human hepatic stellate cells(LX-2) were divided into four groups: blank control group, negative control group, curcumin group(10μM), curcumin + metformin group: curcumin(10μM) + metformin(10 mM). Autophagy related proteins(LC3 B, p62) were detected by Western blot, the rate of cells proliferation was determinated by MTS, the cells apoptosis was detected by flow cytometry instrument. Results Curcumin group could inhibit autophagy and promote apoptosis, and curcumin + metformin group had a stronger synergistic effect with metformin compared with the blank control group and the negative control group, the differences were statistically significant(P<0.05). Conclusion Curcumin and metformin can inhibit autophagy and promote apoptosis of hepatic stellate cells(LX-2).
Objective To investigate the effect of curcumin and metformin on the autophagy and apoptosis of hepatic stellate cells(LX-2). Methods Human hepatic stellate cells(LX-2) were divided into four groups: blank control group, negative control group, curcumin group(10μM), curcumin + metformin group: curcumin(10μM) + metformin(10 mM). Autophagy related proteins(LC3 B, p62) were detected by Western blot, the rate of cells proliferation was determinated by MTS, the cells apoptosis was detected by flow cytometry instrument. Results Curcumin group could inhibit autophagy and promote apoptosis, and curcumin + metformin group had a stronger synergistic effect with metformin compared with the blank control group and the negative control group, the differences were statistically significant(P<0.05). Conclusion Curcumin and metformin can inhibit autophagy and promote apoptosis of hepatic stellate cells(LX-2).
引文
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2杜胜娟,赵伟,夏飞,等.姜黄素的临床应用及其相关作用机制[J].中国现代医药杂志,2017,19(9):93-97
3宋慧东,潘洁,欧阳鹏,等.姜黄素对肝纤维化大鼠中自噬相关蛋白作用的观察[J].新医学,2018,8(49):579-582
4 Bae M,Park YK,Lee JY.Food components with antifibrotic activity and implications in prevention of liver disease[J].J Nutr Biochem,2018,55(5):1-11
5蔡凤.二甲双胍对2型糖尿病大鼠非酒精性脂肪性肝病相关慢性炎症反应的影响[J].实用医学杂志,2018,34(12):1965-1969
6 Nguyen G,Park SY,Le CT,et al.Metformin ameliorates activation of hepatic stellate cells and hepatic fibrosis by succinate and gpr91 inhibition[J].Biochem Biophys Res Commun,2018,495(4):2649-2656
7 Wilson CL,Mann DA,Borthwick LA.Epigenetic reprogramming in liver fibrosis and cancer[J].Adv Drug Deliv Rev,2017,121(11):124-132
8 Munz C.The autophagic machinery in viral exocytosis[J].Front Microbiol,2017,8(2):269
9郜茜,安金霞,李源化,等.肝硬化患者幽门螺杆菌感染和血氨、胃氨关系探讨[J].新医学,2010,41(9):599-601
10 Fu MY,He YJ,Lv X,et al.Transforming growth factorbeta1reduces apoptosis via autophagy activation in hepatic stellate cells[J].Molecular Medicine Reports,2014,10(3):1282-1288
11叶国荣,舒建昌,吕霞,等.姜黄素治疗肝纤维化与肝星状细胞的关系[J].胃肠病学和肝病学杂志,2015,24(10):1227-1231
12 Liu XY,He YJ,Yang QH,et al.Induction of autophagy and apoptosis by mir-148a through the sonic hedgehog signaling pathway in hepatic stellate cells[J].Am J Cancer Res,2015,5(9):2569-2589
13 Li Z,Ding Q,Ling LP,et al.Metformin attenuates motility,contraction,and fibrogenic response of hepatic stellate cells in vivo and in vitro by activating amp-activated protein kinase[J].World J Gastroenterol,2018,24(7):819-832
14邓金鹏.熊去氧胆酸联合二甲双胍对非酒精性脂肪肝患者肝功能的影响[J].临床医学研究与实践,2018,8(3):40-41
15易思妮,颜灿群.姜黄素与糖尿病认知功能障碍的研究进展[J].中国现代医药杂志,2016,18(12):97-99
16 Ko MT,Huang HC,Lee WS,et al.Metformin reduces intrahepatic fibrosis and intrapulmonary shunts in biliary cirrhotic rats[J].Journal of The Chinese Medical Association,2017,80(8):467-475
17 Fan K,Wu K,Lin L,et al.Metformin mitigates carbon tetrachloride-induced tgf-beta1/smad3 signaling and liver fibrosis in mice[J].Biomed Pharmacother,2017,90(6):421-426
18 Doyle MA,Singer J,Lee T,et al.Improving treatment and liver fibrosis outcomes with metformin in hcv-hiv co-infected and hcv mono-infected patients with insulin resistance:study protocol for a randomized controlled trial[J].Trials,2016,17(1):331
19 Barzilai N,Crandall JP,Kritchevsky SB,et al.Metformin as a tool to target aging[J].Cell Metab,2016,23(6):1060-1065