大黄素甲醚对大鼠肝星状细胞HSC-T6细胞增殖及凋亡的影响
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摘要
目的初步探讨大黄素甲醚对HSC-T6细胞增殖及凋亡的影响。方法采用溶剂系统分离法从虎杖中制备出单体大黄素甲醚(Physcion);Cell Counting Kit-8(CCK-8试剂盒)法检测大黄素甲醚对HSC-T6细胞存活率的影响;通过Ed U染色法进一步观察不同浓度(5、10、20μmol/L)大黄素甲醚作用后,HSC-T6细胞的增殖状况;免疫印迹法(Western blot)检测Bcl-2、Bax蛋白的表达变化。结果与对照组(加入等量的不含药物的培养基)相比,大黄素甲醚可抑制HSC-T6细胞增殖,且呈浓度依赖性;Ed U染色可观察到大黄素甲醚组阳性细胞数目及所占比例均显著小于对照组(P<0.001);Western blot结果显示大黄素甲醚在10μmol/L和20μmol/L时可降低Bcl-2表达水平(P<0.001),同时升高Bax水平(P<0.001)。结论大黄素甲醚可抑制HSC-T6细胞增殖,可能是通过调节Bax/Bcl-2水平来发挥其作用。
Objective To study the effect of physcion on proliferation and apoptosis of rat hepatic stellate cells( HSC-T6 cell). Methods Physcion was prepared from Polygonum cuspidatum by solvent system separation method.CCK-8 assay was used to test the cell viability. Edu stain were used to assess proliferation of HSC-T6 cells. Western Blot assay was used to assess the expression of Bcl-2 and Bax. Results Compared with control group,physcion could inhibit the proliferation of HSC-T6 cells in a concentration-dependent manner. The number and rate of edu positive cells were significantly less in physcion group than those in control group( P < 0. 001). Western Blot showed that the expression of Bcl-2 was significantly reduced by physcion at 10 ed t/L and 20 ion/L; whereas Bax was significantly increased. Conclusion Physcion inhibits cell proliferation of HSC-T6,which may via adjusting Bax/Bcl-2 levels.
Objective To study the effect of physcion on proliferation and apoptosis of rat hepatic stellate cells( HSC-T6 cell). Methods Physcion was prepared from Polygonum cuspidatum by solvent system separation method.CCK-8 assay was used to test the cell viability. Edu stain were used to assess proliferation of HSC-T6 cells. Western Blot assay was used to assess the expression of Bcl-2 and Bax. Results Compared with control group,physcion could inhibit the proliferation of HSC-T6 cells in a concentration-dependent manner. The number and rate of edu positive cells were significantly less in physcion group than those in control group( P < 0. 001). Western Blot showed that the expression of Bcl-2 was significantly reduced by physcion at 10 ed t/L and 20 ion/L; whereas Bax was significantly increased. Conclusion Physcion inhibits cell proliferation of HSC-T6,which may via adjusting Bax/Bcl-2 levels.
引文
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[2]廖文,李明.虎杖的本草考证及药理作用[J].中华中医药学刊,2011,29(11):2511-2513.
[3]夏海珊,陈少茹,钟月春,等.肝纤维化的发病机制和药物治疗现况[J].中国医药导报,2014,11(18):162-165.
[4]杨悦杰,黄芬.肝星状细胞及相关细胞因子在肝纤维化形成中的作用[J].世界华人消化杂志,2007,15(27):2885-2890.
[5]Seki E,Brenner DA.Recent advancement of molecular mechanisms of liver fibrosis[J].J Hepatobiliary Pancreat Sci,2015,22(7):512-518.
[6]Trautwein C,Friedman SL,Schuppan D,et al.Hepatic fibrosis:Concept to treatment[J].J Hepatol,2015,62(1 Suppl):S15-S24.
[7]丁诗正,骆传佳.肝纤维化的中医药治疗概况及其特点[J].中国中医药咨讯,2011,3(23):289.
[8]高晚霞,罗丽丹,陈舒丽,等.虎杖对大鼠急性四氯化碳肝损伤保护作用的量效关系[J].咸宁学院学报,2007,21(3):215-217.
[9]吴德跃,吴俊标,周玖瑶,等.虎杖水提液利胆保肝作用研究[J].西北药学杂志,2014,29(2):167-169.
[10]杨先振,赵有亮,秦红兵.中药虎杖水煎液对CCL4诱导肝纤维化的保护作用[J].江苏医药,2011,37(23):2761-2763.
[11]陈剑明,张声生,吴震宇,等.虎杖苷对非酒精性脂肪肝保护作用的实验研究[J].中华中医药学刊,2015,33(5):1188-1191.
[12]贾玉梅,王君明,崔瑛.基于二苯乙烯类为主要活性成分的虎杖药理作用研究进展[J].中国实验方剂学杂志,2011,19(9):263-269.
[13]杨桂智,赵心怡,黄敏霞,等.虎杖苷对肝纤维化治疗作用实验研究[J].亚太传统医药,2017,13(16):14-16.
[14]张霖,陈育尧,孙学刚,等.虎杖苷对非酒精性脂肪肝大鼠保护作用及机制研究[J].陕西中医,2010,31(6):756-758.
[15]蔺红伟,江春霞,朴淑娟.用HPLC法测定不同地区虎杖饮片中的4种有效成分[J].药学服务与研究,2017,17(1):27-30.
[16]张丽雁,宿文辉,熊瑛.大黄素甲醚对大鼠急性肝损伤的保护作用[J].中国基层医药,2014,14(12):2014-2015.
[17]Novo E,Marra F,Zamara E,et al.Overexpression of Bcl-2 by activated human hepatic stellate cells:resistance to apoptosis as a mechanism of progressive hepatic fibrogenesis in humans[J].Gut,2006,55(8):1174-1182.
[18]González-Puertos VY,Hernández-Pérez E,Nuňo-Lámbarri N,et al.Bcl-2 overexpression in hepatic stellate cell line CFSC-2G,induces a pro-fibrotic state[J].J Gastroenterol Hepatol,2010,25(7):1306-1314.