摘要
目的:探讨天胡荽异牡荆素对大鼠肝星状细胞(HSC-T6)增殖、凋亡以及磷脂酸肌醇-3-激酶/蛋白激酶B(PI3K/Akt)信号通路的影响。方法:将HSC-T6细胞分为正常对照组、模型组、阳性药(LY294002)组和异牡荆素高、中、低剂量组(120μmol/L、60μmol/L、30μmol/L)。血小板衍生生长因子BB(PDGF-BB)刺激HSC-T6细胞使其活化,各组给与相应药物进行干预24 h,采用CCK-8法检测异牡荆素对HSC-T6细胞增殖的影响;吖啶橙/溴化乙锭(AO/EB)荧光染色实验观察细胞凋亡形态学的变化;流式细胞术检测细胞凋亡率的变化;qPCR和Western blot法检测各组细胞中α-平滑肌肌动蛋白(α-SMA)、PI3K、p-PI3K、Akt和p-Akt基因和蛋白表达水平。结果:与正常对照组相比,PDGF-BB刺激后HSC-T6细胞增殖明显(P<0.01);同时,细胞内α-SMA、PI3K、p-PI3K、Akt和p-Akt基因和蛋白的表达均显著增加(P<0.01)。与模型组相比,异牡荆素能够明显抑制HSC-T6细胞增殖,诱导细胞凋亡(P<0.01)。此外,异牡荆素干预能够显著减少肝纤维化标志蛋白α-SMA的表达并抑制胞内PI3K和Akt蛋白磷酸化水平(P<0.01)。结论:天胡荽异牡荆素能够抑制活化HSC-T6细胞增殖,诱导细胞凋亡,其机制可能与抑制PI3K/Akt信号通路有关。
Objective:To discuss the effects of isovitexin in hydrocotyle sibthorpioides on the proliferation and apoptosis of rat hepatic stellate cells(HSC-T6)and the PI3K/Akt signaling pathway.Methods:HSC-T6 cells were divided into control group,model group,positive drug(LY294002)group,and groups of high,medium,and low dose of isovitexin(120μmol/L,60μmol/L,30μmol/L).Except for the control group,the rest of the cell groups were activated by platelet-derived growth factor BB(PDGF-BB)then treated with corresponding drugs for 24 h.The effect of isovitexin on the proliferation of HSC-T6 cells was measured by Cell Counting Kit-8(CCK-8).Acridine orange/Ethidium bromide(AO/EB)fluorescence staining was used to observe changes in cell apoptosis morphology.Flow cytometry was used to detect cell apoptotic rate.Western blot and real-time quantitative polymerase chain reaction(qPCR)were used to detect the expression levels ofα-smooth muscle Kinesin(α-SMA),PI3K,p-PI3K,Akt and p-Akt.Results:Compared with the control group,cell proliferation was significantly increased in the rest of the cell groups after PDGF-BB stimulation(P<0.01);meanwhile,the expression level ofα-SMA,PI3K,p-PI3K,Akt,and p-Akt were significantly increased(P<0.01).Comparison between isovitexin groups and the model group suggested that isovitexin could significantly inhibit the proliferation of HSC-T6 cells and induce apoptosis(P<0.01).In addition,isovitexin could significantly reduce the expression of liver fibrosis marker proteinα-SMA and inhibite phosphorylation of PI3K and Akt(P<0.01).Conclusion:Isovitexin in hydrocotyle sibthorpioides can inhibit the proliferation of activated HSC-T6 cells and induce cell apoptosis.The mechanism may be related to the inhibition of PI3K/Akt signaling pathway.
作者
Vonglorkham Sayyaphone
乔倩
庞小红
王丹丹
冯钟文
庞丽君
陈思韵
韦锦斌
Vonglorkham Sayyaphone;Qiao Qian;Pang Xiaohong;Wang Dandan;Feng Zhongwen;Pang Lijun;Chen Siyun;Wei Jinbin(Pharmaceutical College,Guangxi Medical University,Nanning 530021,China;The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2020年第2期182-187,共6页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No.81960767)
广西教育厅“药学研究生创新创业教育暨联合培养基地项目”(No.20170703)。
