蓝莓花青素对氧化应激诱导肝星状细胞活化、增殖和凋亡的影响及机制探讨  

Effects of blueberry anthocyanins on activation and proliferation of oxidative stress-induced rat hepatic stellate cells

作　　者：王丽[1] 何萍[1] 孙建超[1] 王豫萍[1] 程明亮[1] 

机构地区：[1]贵州医科大学临床检验学院,贵阳550004

出　　处：《山东医药》2016年第18期8-11,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81460125);贵州省留学人员科技活动项目(20151010);贵州省科技厅中药现代项目(黔科合ZY字-2012-3017);贵州省省长基金项目(黔省专合字2012-44)

摘　　要：目的观察蓝莓提取物花青素对氧化应激诱导肝星状细胞HSC-T6活化、增殖的影响,并探讨其作用机制。方法取对数生长期HSC-T6细胞分为观察组、对照组、空白组,观察组先加入600μg/m L蓝莓花青素干预24h后,加葡萄糖氧化酶(GO)100 U/L孵育3 h,对照组加入GO 100 U/L孵育3 h,空白组不做任何处理。干预24 h时采用细胞免疫化学法检测各组α-平滑肌肌动蛋白(α-SMA),ELISA法检测胞质Ⅰ型胶原(ColⅠ);采用MTT法观察各组细胞增殖情况,采用流式细胞仪观察各组细胞凋亡情况;采用硫代巴比妥酸法检测各组胞质丙二醛(MDA),黄嘌呤氧化酶法检测各组胞质SOD,微量酶标法测定各组胞质谷胱甘肽(GSH)。结果干预24 h时,各组α-SMA相对表达量为空白组<观察组<对照组,P均<0.05。干预24 h时,观察组细胞增殖率低于对照组,P<0.05。干预24 h时,观察组细胞凋亡率高于其余两组,P均<0.05。干预24 h时,观察组胞质MDA低于对照组(P<0.05),SOD、GSH均高于空白组(P均<0.05)。结论蓝莓花青素可抑制氧化应激诱导的肝星状细胞的活化与增殖,并促进细胞凋亡;其机制可能与抑制胞质内MDA表达,促进SOD、GSH表达有关。Objective To investigate the effects of blueberry anthocyanins on the activation, proliferation and apopto- sis of oxidative stress-induced rat hepatic stellate cells （HSC-T6） and the correlated mechanisms. Methods HSC-T6 cells in the logarithmic phase Were divided into the observation group, control group and the blank group, respectively. The ob- servation group was incubated with 100 μg/mL U/L GO for 3 hours after blueberry anthocyanin intervention of 24 hours, the control group was incubated with 100 U/L GO for 3 hours, and the blank group without any treatment. After 24-hour in- tervention, the expression of cL-smooth muscle actin （ct-SMA） in HSC-T6 cells was measured by immunohistochemistry, the cytosohc collagen I （Col I） level was detected by ELISA, the cell proliferation was detected by MTT, the apoptosis of HSC-T6 was detected by flow cytometry, the level of MDA was measured by thiobarbituric acid method, and the activity of superoxide dismutase （SOD） and the level of GSH were detected by xanthine oxidase and enzyme linked immunosorbent as- say, respectively. Results After intervention of 24 h, the expression level of α-SMA in each group was blank group 〈 ob- servation group 〈 control group, all P 〈 0.05. The HSC-T6 proliferation rate of the observation group was significantly lower than that in the control group, P 〈 0.05. Nevertheless, the apoptosis rate of the observation group was significantly higher than those of the other groups, all P 〈 0.05. After 24-hour intervention, the level of MDA in the observation group was de- creased significantly as compared with that of the control group, P 〈 0.05. The levels of SOD and GSH in the observation group were all higher than those of the blank group all P 〈 0.05. Condusion Blueberry anthocyanins can significantly in- hibit the activation and proliferation and promote the apoptosis of HSC, and the mechanisms may be related to inhibiting MDA and promoting the expression of SOD and GSH.

关 键 词：肝星状细胞 蓝莓 花青素 氧化应激 Α-平滑肌肌动蛋白 Ⅰ型胶原 细胞增殖 细胞凋亡 

分 类 号：R575.2[医药卫生—消化系统]