珠子参总皂苷通过上调miR-325-3p抑制脂多糖诱导的星形胶质细胞凋亡和炎症因子释放  

Total Saponins from Panacis Majoris Rhizoma Inhibiting Lipopolysaccharide-induced Astrocyte Apoptosis and Inflammatory Factor Release by Up-regulating miR-325-3p

作　　者：周威 黄贤亮 吴旭明 Zhou Wei;Huang Xianliang;Wu Xuming(Department of Phamiacy,First People's Hospital of Huzhou,Zhejiang Huzhou 313000,China)

机构地区：[1]湖州市第一人民医院药剂科,浙江湖州313000

出　　处：《中国药师》2021年第9期1617-1622,1635,共7页China Pharmacist

基　　金：浙江省湖州市科学技术局项目(编号:2016GYB18)。

摘　　要：目的:探讨珠子参总皂苷对脂多糖诱导的星形胶质细胞凋亡和炎症因子表达的影响及可能机制。方法:体外培养大鼠星形胶质细胞,不同剂量(50,100,200μg·ml-1)的珠子参总皂苷作用于脂多糖诱导的星形胶质细胞24 h、脂多糖处理转染miR-325-3p模拟物的星形胶质细胞24 h或200μg·ml-1的珠子参总皂苷作用于脂多糖诱导的的转染miR-325-3p抑制剂的星形胶质细胞24 h后,流式细胞术检测细胞凋亡,蛋白印迹法检测细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved-caspase3)和活化的含半胱氨酸的天冬氨酸蛋白水解酶9(cleaved-caspase9)蛋白表达,试剂盒检测细胞培养上清中肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)水平,实时定量聚合酶链式反应(RT-qPCR)检测细胞中微小RNA-325-3p(miR-325-3p)表达。结果:珠子参总皂苷可降低脂多糖诱导的星形胶质细胞凋亡率、细胞中cleaved-caspase3和cleaved-caspase9蛋白表达及TNF-α和IL-6的分泌(P<0.05),促进miR-325-3p表达(P<0.05),且呈剂量依赖性。过表达miR-325-3p可降低脂多糖诱导的星形胶质细胞凋亡率、细胞中cleaved-caspase3和cleaved-caspase9蛋白表达及TNF-α和IL-6的分泌(P<0.05)。敲减miR-325-3p逆转珠子参总皂苷对脂多糖诱导的星形胶质细胞凋亡及TNF-α和IL-6表达的抑制作用(P<0.05)。结论:珠子参总皂苷可能通过上调miR-325-3p表达抑制脂多糖诱导的大鼠星形胶质细胞凋亡和炎症反应。Objective:To investigate the effects of total saponins from Panacis Majoris Rhizoma on lipopolysaccharide-induced astrocyte apoptosis,inflammatory factor expression and its possible mechanism.Methods:Rat astrocytes were cultured in vitro.After different doses(50,100,200μg·ml-1)of total saponins from Panacis Majoris Rhizoma acted on lipopolysaccharide-induced astrocytes for 24 h,lipopolysaccharide treated astrocytes were transfected with miR-325-3 p mimic for 24 h or 200μg·ml-1 total saponins from Panacis Majoris Rhizoma acted on lipopolysaccharide-induced astrocytes were transfected with miR-325-3 p inhibitor for 24 h,and then flow cytometry was used to detect cell apoptosis,Western blotting was used to detect the protein expressions of cleaved-caspase3 and cleaved-caspase9 in cells,kits were used to detect the levels of TNF-αand IL-6 in cell culture supernatants,and RT-qPCR was used to detect the expression of miR-325-3 p.Results:Total saponins from Panacis Majoris Rhizoma could reduce the apoptosis rate of astrocytes induced by lipopolysaccharide,and decrease the protein expressions of cleaved-caspase3 and cleaved-caspase9 in cells,and the secretion of TNF-αand IL-6 as well(P<0.05),while promote the expression of miR-325-3 p(P<0.05)in a dose-dependent manner.Overexpression of miR-325-3 p could reduce the apoptosis rate of astrocytes induced by lipopolysaccharide,and decrease the protein expressions of cleaved-caspase3 and cleaved-caspase9 in cells,and the secretion of TNF-αand IL-6 as well(P<0.05).Knockdown of miR-325-3 p reversed the inhibitory effects of total saponins from Panacis Majoris Rhizoma on the apoptosis of lipopolysaccharide-induced astrocyte and the expressions of TNF-αand IL-6(P<0.05).Conclusion:Total saponins from Panacis Majoris Rhizoma may inhibit lipopolysaccharide-induced apoptosis and inflammation of rat astrocytes by up-regulating the expression of miR-325-3 p.

关 键 词：珠子参总皂苷 星形胶质细胞 微小RNA-325-3p 细胞凋亡 炎症 

分 类 号：R285.5[医药卫生—中药学]