羊栖菜多糖通过miR-7抑制MPP+诱导的SK-N-SH损伤  

Effects of Sargassum fusiforme polysaccharide on SK-N-SH injury induced by MPP+through miR-7

作　　者：李茜[1] 李永秋[1] 张冬森[1] 高轩[1] 高海凤[1] LI Xi;LI Yongqiu;ZHANG Dongsen;GAO Xuan;GAO Haifeng(Department of Neurology,Tangshan Workers′Hospital,Tangshan 063000,China)

机构地区：[1]唐山市工人医院神经内科,河北唐山063000

出　　处：《沈阳药科大学学报》2023年第11期1449-1455,共7页Journal of Shenyang Pharmaceutical University

摘　　要：目的羊栖菜多糖(SFPS)对1-甲基-4-苯基吡啶离子(MPP+)诱导的SK-N-SH损伤的影响及分子机制。方法将SK-N-SH细胞用0.3 mmol·L^(-1)的MPP+处理作为模型(Model)组;以正常培养的细胞作为对照(Con)组,用质量浓度分别为10、30、100 mg·L^(-1)的羊栖菜多糖和0.3 mmol·L^(-1)的MPP+处理作为羊栖菜多糖低、中、高浓度组;将SK-N-SH细胞再分为Model+miR-NC组、Mod-el+miR-7组、Model+SFPS-H+anti-miR-NC组、Model+SFPS-H+anti-miR-7组;实时荧光定量PCR(RT-qPCR)检测miR-7表达水平;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测蛋白表达;试剂盒检测细胞MDA含量、SOD活性和培养液中LDH含量。结果不同浓度羊栖菜多糖处理后,MPP+诱导的SK-N-SH细胞中miR-7表达水平升高,细胞的凋亡率降低,Bax表达水平降低,Bcl-2表达水平升高,MDA含量和LDH水平降低,SOD活性升高。过表达miR-7后,MPP+诱导的SK-N-SH细胞的凋亡率降低,Bax表达水平降低,Bcl-2表达水平升高,MDA含量和LDH水平降低,SOD活性升高。抑制miR-7逆转了羊栖菜多糖对MPP+诱导的SK-N-SH细胞损伤的作用。结论羊栖菜多糖可能通过上调miR-7抑制MPP+诱导的SK-N-SH细胞损伤。Objective To explore the effects and molecular mechanism of Sargassum fusiforme polysaccharide(SFPS)on 1-methyl-4-phenylpyridinium(MPP+)-induced SK-N-SH damage.Methods The SK-N-SH cells were treated with 0.3 mmol·L^(-1)MPP+as the model group;normal cultured cells were used as the control(Con)group,with concentrations of 10 mg·L^(-1),30 mg·L^(-1),and 100 mg·L^(-1)of Sargassum fusiforme polysaccharide and 0.3 mmol·L^(-1)of MPP+treatment as the low,medium and high concentration Sargassum fusiforme polysaccharide groups;SK-N-SH cells were divided into Model+miR-NC group and Model+miR-7 group,Model+SFPS-H+anti-miR-NC group,Model+SFPS-H+anti-miR-7 group;The real-time fluorescent quantitative PCR(RT-qPCR)was used to detect miR-7 expression level;The flow cytometry was used to detect cells apoptosis;Western blot method was employed to detect protein expression;The cell MDA content,SOD activity and LDH content were detected by kits in culture medium.Results After treatment with different concentrations of Sargassum fusiforme polysaccharide,the expression level of miR-7 in SK-N-SH cells induced by MPP+was increased,the cells apoptosis rate was decreased,the expression level of Bax was decreased,the expression level of Bcl-2 was increased,the levels of MDA and LDH were decreased,and the SOD activity was increased(P<0.05).After overexpression of miR-7,the apoptosis rate of SK-N-SH cells induced by MPP+was decreased,the expression level of Bax was decreased,the expression level of Bcl-2 was increased,the levels of MDA and LDH were decreased,and the activity of SOD was increased(P<0.05).Inhibition of miR-7 reversed the effect of Sargassum fusiforme polysaccharide on MPP+-induced SK-N-SH cell damage.Conclusion Sargassum fusiforme polysaccharide may inhibit MPP+-induced SK-N-SH cell damage by up-regulating miR-7.

关 键 词：羊栖菜多糖 miR-7 SK-N-SH细胞 帕金森病 

分 类 号：R285.5[医药卫生—中药学] R96[医药卫生—中医学]