中国中药杂志

2018, v.43(02) 390-395

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竹节参总皂苷对棕榈酸诱导HepG2细胞凋亡的保护作用研究
Protective effects of total saponins of Panax japonicas on HepG2 cell apoptosis induced with palmitic acid

涂浩;周琴;张迅;王婷;袁成福;张长城;袁丁;刘朝奇;
TU Hao;ZHOU Qin;ZHANG Xun;WANG Ting;YUAN Cheng-fu;ZHANG Chang-cheng;YUAN Ding;LIU Zhao-qi;Medical College,Three Gorges University;Renhe Hospital,Three Gorges University;

摘要(Abstract):

探讨竹节参总皂苷(TSPJ)对棕榈酸(PA)诱导的HepG2细胞凋亡的保护作用及分子机制。体外培养HepG2细胞,分为5组:对照组,模型组,竹节参参总皂苷高(50 mg·L~(-1))、中(25 mg·L~(-1))、低(12.5 mg·L~(-1))剂量组。5组细胞均培养24 h后,采用MTT法检测细胞活力,用Hoechst染色和Annexin V-FITC/PI双标记流式细胞术检测细胞凋亡,用RT-PCR法检测TNF-α,IL-1β,BCL-2,CHOP和GAPDH mRNA表达水平,用Western blot法和流式检测BCL-2,CHOP和Toll样受体4(TLR4)蛋白表达量。结果表明,与正常组比较,模型组细胞数量显著减少(P<0.01),凋亡细胞增多,与模型组比较,高,中剂量组细胞数量显著增多(P<0.01),凋亡细胞减少;与正常组比较,模型组CHOP和TLR4蛋白表达量,TNF-α,IL-1β和CHOP mRNA表达水平显著升高(P<0.01),BCL-2蛋白表达水平和mRNA表达水平显著降低(P<0.01),与模型组比较,高剂量组CHOP和TLR4蛋白表达量,TNF-α,IL-1β和CHOP mRNA表达水平显著降低(P<0.01),BCL-2蛋白表达量和mRNA表达水平显著升高(P<0.01),提示竹节参总皂苷可以减轻PA诱导的炎症反应及凋亡,对肝细胞有一定的保护作用。
This study aimed to investigate the molecular mechanism and protective effect of total saponins of Panax japonicas(TSPJ) on HepG2 cells apoptosis induced by palmitic acid( PA). The HepG2 cells were cultured in vitro,and divided into five groups: the control group,the model group,the high-dose group( 50 mg·L~(-1)),the middle-dose group(25 mg·L~(-1)) and the lowdose group( 12. 5 mg·L~(-1)). The cells of the five groups were cultured continuously for 24 hours. The cell viability was measured with MTT. HepG2 cells apoptosis was detected by Hoechest staining and Annexin V-FITC/PI staining. The protein expressions of BCL-2,CHOP and TLR4 were measured with western blotting and flow cytometry analysis. The mRNA expressions of TNF-α,IL~(-1) β,BCL-2,CHOP and GAPDH were measured with RT-PCR. The results suggested that compared with the control group,the number of HepG2 cells of the model group were reduced significantly( P < 0. 01),while the number of apoptotic HepG2 cells were increased. Compared with the model group,the number of HepG2 cells of the high-dose group and the middle-dose group were increased significantly( P <0. 01),whereas the number of apoptotic HepG2 cells were reduced. Compared with the control group,TNF-α,IL~(-1) β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the model group were significantly up-regulated( P < 0. 01),while BCL-2 protein and mRNA expressions in the model group were significantly decreased( P < 0. 01). Compared with the model group,TNF-α,IL~(-1) β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the high-dose group were significantly decreased( P <0. 01),while BCL-2 protein and mRNA expressions in the high-dose group were significantly up-regulated( P < 0. 01). In conclusion,TSPJ can reduce inflammation and apoptosis induced by palmitic acid,with a certain protective effect on liver cells.

关键词(KeyWords): 竹节参总皂苷;棕榈酸;凋亡;内质网应激;非酒精性脂肪肝炎
total saponins of Pana japonicas(TSPJ);palmitic acid(PA);apoptosis;endoplasmic reticulum stress(ERS);non-alcoholic steatohepatitis(NASH)

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81673675);; 三峡大学研究生科研创新基金项目(SDYC2016092)

作者(Author): 涂浩;周琴;张迅;王婷;袁成福;张长城;袁丁;刘朝奇;
TU Hao;ZHOU Qin;ZHANG Xun;WANG Ting;YUAN Cheng-fu;ZHANG Chang-cheng;YUAN Ding;LIU Zhao-qi;Medical College,Three Gorges University;Renhe Hospital,Three Gorges University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20171027.011

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