活血解毒方对HSG细胞凋亡中Caspase家族及关键蛋白的影响  

Effects of Huoxue Jiedu Decoction on Caspase family and key proteins in HSG cell apoptosis

作　　者：张赛[1] 沈正东 郑凌琦 徐江喜 张玉婷[1] 刘小平[2] 侯秀娟[2] 朱跃兰[2] ZHANG Sai;SHEN Zhengdong;ZHENG Lingqi;XU Jiangxi;ZHANG Yuting;LIU Xiaoping;HOU Xiujuan;ZHU Yuelan(Beijing University of Chinese Medicine,Beijing 100029,China;Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学东方医院,北京100078

出　　处：《中华中医药杂志》2023年第7期3310-3313,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81874438)。

摘　　要：目的:观察活血解毒方对HSG细胞凋亡中Caspase家族及关键蛋白的影响。方法:IFN-γ+TRAIL联合诱导的HSG细胞凋亡,活血解毒方作为中药组药物干预,以白芍总苷(TGP)作为阳性对照组,实时荧光定定量(qPCR)检测各组Caspase(3,8,9)mRNA表达,免疫组化(IHC)检测各组Caspase(3,8,9)蛋白OD值,基因沉默Caspase-8,流式细胞术检测各组细胞Caspase-8的凋亡率。结果:与空白组比较,模型组Caspase(3,8,9)mRNA相对表达量及蛋白OD值显著升高(P<0.01);与模型组比较,TGP组和中药组Caspase(3,8,9)mRNA相对表达量均显著降低,Caspase-3蛋白OD值均显著降低(P<0.01),中药组Caspase(8,9)蛋白OD值显著降低(P<0.01,P<0.05);与TGP组比较,中药组Caspase(3,8,9)mRNA相对表达量显著降低(P<0.01);与空白组比较,siRNA NC和siRNA Caspase-8组HSG细胞凋亡率显著升高(P<0.01);与siRNA NC组比较,siRNA-Caspase-8组HSG细胞凋亡率显著升高(P<0.01);与siRNA NC+IFN-γ+TRAIL+中药组比较,siRNA-Caspase-8+IFN-γ+TRAIL组HSG细胞凋亡率显著降低(P<0.01);与siRNA-Casepase-8+IFN-γ+TRAIL组比较,siRNA-Caspase-8+IFN-γ+TRAIL+中药组HSG细胞凋亡率显著降低(P<0.01)。结论:活血解毒方能有效抑制Caspase家族细胞凋亡相关因子,这一过程可能是通过下调Caspase-8的表达实现的。Objective:To observe the effects of Huoxue Jiedu Decoction on Caspase family and key proteins in HSG cell apoptosis.Methods:The apoptosis of HSG cells induced by the combination of IFN-γ+TRAIL was treated with Huoxue Jiedu Decoction as the TCM group,and total glucosides of paeony(TGP)as the positive control group.The qPCR detection of Caspase(3,8,9)mRNA expression,IHC detection of Caspase(3,8,9)protein OD value,gene silencing Caspase-8,and flow cytometry detection of the apoptosis rate of Caspase-8 were used in each group.Results:Compared with the blank group,the relative expression levels of Caspase(3,8,9)mRNA and protein OD values in the model group were significantly increased(P<0.01).Compared with the model group,the relative expression of Caspase(3,8,9)mRNA in the TGP group and the TCM group was significantly decreased(P<0.01),the OD value of Caspase-3 protein was significantly decreased(P<0.01),and the OD value of Caspase(8,9)protein in the TCM group was significantly decreased(P<0.01).Compared with the TGP group,the relative expression of Caspase(3,8,9)mRNA in the TCM group was significantly decreased(P<0.01).Compared with the blank group,the apoptosis rate of HSG cells in the siRNA NC and siRNA Caspase-8 groups was significantly increased(P<0.01).Compared with siRNA NC group,the apoptosis rate of HSG cells in siRNA-Caspase-8 group was significantly increased(P<0.01).Compared with siRNA NC+IFN-γ+TRAIL+TCM group,the apoptosis rate of HSG cells insiRNA Caspase-8+IFN-γ+TRAIL group was significantly decreased(P<0.01).Compared with siRNA-Casepase-8+IFN-γ+TRAIL group,the apoptosis rate of HSG cells in siRNA-Caspase-8+IFN-γ+TRAIL+TCM group was significantly lower(P<0.01).Conclusion:Huoxue Jiedu Decoction can effectively inhibit the apoptosis-related factors of Caspase family,which may be achieved by down-regulating the expression of Caspase-8.

关 键 词：干燥综合征 HSG细胞 活血解毒方 细胞凋亡 CASPASE家族 CASPASE-8 

分 类 号：R285[医药卫生—中药学]