摘要
目的:研究棒柄花苷A(PPGA)对白介素-1β(IL-1β)诱导活化的大鼠肝星状细胞(HSC-T6)增殖、胶原、凋亡和细胞周期的影响。方法:通过MTT法检测PPGA对HSC-T6细胞活力的影响,计算半数抑制浓度(IC_(50)),以IC_(50)设置给药浓度。将HSCT6细胞分为正常对照组、模型组和PPGA高、中、低剂量组,除正常对照组外,其他各组均用IL-1β刺激30 min使HSC-T6细胞活化,建立体外肝纤维化模型。药物干预24 h后,采用MTT法检测HSC-T6细胞增殖活力,免疫荧光法检测Ⅰ型胶原蛋白(Col-Ⅰ)和Ⅲ型胶原蛋白(Col-Ⅲ)表达,流式细胞仪检测细胞凋亡和细胞周期。结果:PPGA作用于HSC-T6细胞24 h、48 h、72 h的IC_(50)分别为473 mg/L、100 mg/L、15 mg/L,因此选择200 mg/L、100 mg/L、50 mg/L分别作为PPGA高、中、低剂量进行后续实验。与正常对照组比较,模型组HSC-T6细胞增殖能力提高,Col-Ⅰ和Col-Ⅲ蛋白表达升高(均P<0.05),而细胞凋亡率和周期分布无明显改变(P>0.05);与模型组比较,PPGA高、中剂量组HSC-T6细胞增殖活力降低,Col-Ⅲ蛋白表达降低,PPGA各剂量组Col-Ⅰ蛋白表达降低,凋亡率升高,细胞S期延长(均P<0.05)。结论:PPGA可抑制HSC-T6细胞增殖,诱导细胞凋亡,并使HSC-T6细胞周期阻滞在S期,减缓肝纤维化的发展进程。
Objective:To investigate the effects of PPGA from Cleidion brevipetiolatum Paxet Hoffm on proliferation,collagen,apoptosis and cell cycle of activated rat hepatic stellate cells(HSC-T6)induced by interleukin-1β(IL-1β).Methods:The effect of PPGA on the viability of HSC-T6 cells was detected by MTT method.The 50%inhibitory concentration(IC_(50))was calculated.The drug concentration was set by IC_(50).HSC-T6 cells were divided into normal control group,model group and high,middle and low dose groups of PPGA.Except for the normal control group,HSC-T6 cells were activated by IL-1βstimulation for 30 min to establish an in vitro model of liver fibrosis.After 24 hours of drug intervention,the proliferation activity of HSC-T6 cells was detected by MTT method,the expression of typeⅠcollagen(Col-Ⅰ)and typeⅢcollagen(Col-Ⅲ)was detected by immunofluorescence,and the apoptosis and cell cycle were detected by flow cytometry.Results:The IC_(50) of HSC-T6 cells treated with PPGA for 24 h,48 h and 72 h was 473 mg/L,100 mg/L and 15 mg/L,respectively,so 200 mg/L,100 mg/L and 50 mg/L were selected as high,middle and low doses of PPGA for follow-up experiments.Compared with the normal control group,the proliferation ability of HSC-T6 cells and the expression of Col-Ⅰand Col-Ⅲproteins in the model group increased(all P>0.05),while the apoptosis rate and cell cycle distribution had no significant change(P>0.05).Compared with the model group,the proliferation activity of HSC-T6 cells and Col-Ⅲprotein expression in the high and middle dose groups of PPGA decreased,and the expression of Col-Ⅰprotein decreased,the apoptosis rate increased and the S phase prolonged in all PPGA groups(all P<0.05).Conclusion:PPGA can inhibit the proliferation of HSC-T6 cells,induce apoptosis,block the HSC-T6 cell cycle in S phase,and slow down the development of liver fibrosis.
作者
黄奕璇
蒋莉夏
何云
刘元
杨斌
黄春英
Huang Yixuan;Jiang Lixia;He Yun;Liu Yuan;Yang Bin;Huang Chunying(Pharmaceutical College of Guangxi Medical University,Nanning 530021,China;Traditional Chinese Medicine Hospital of Yulin,Yulin 537000,China;Guangxi Institute of Chinese Medicine&Pharmaceutical Science,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2022年第6期886-891,共6页
Journal of Guangxi Medical University
基金
广西自然科学基金资助项目(No.2018GXNSFAA050125)。
关键词
棒柄花苷A
肝星状细胞
肝纤维化
增殖
凋亡
PPGA
hepatic stellate cell
hepatic fibrosis
proliferation
apoptosis
