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目的:此实验设计以了解羊栖菜多糖(Sargassum Fusiforme Polysaccharides,SFPS)对胰岛β细胞的保护作用及机制。方法:采用体外H2O2诱导大鼠胰岛细胞瘤株(RINm5F)凋亡,MTT法检测SFPS对RINm5F的活性影响,运用分光光度法检测SFPS对RINm5F和原代大鼠胰岛的SOD、MDA的影响。结果:0.2mmol/LH2O2诱导RINm5F凋亡,Hoechst3342/PI见典型的凋亡,30μmol/LPI3K抑制剂LY294002与H2O2有协同致细胞损伤作用;20μg.ml-1SFPS保护H2O2对RINm5F活性的损伤,LY294002预处理1h不能阻断其保护作用;模型组RINm5F和原代大鼠胰岛细胞内SOD下降;SFPS能提高RINm5F和原代大鼠胰岛的SOD活性,LY294002预处理阻断SFPS提高抗氧化酶活性的作用。结论:H2O2可导致胰岛细胞凋亡,使细胞和组织的抗氧化酶活性下降,与PI3K-Akt通路抑制剂有协同作用,一定浓度和时间SFPS可减少H2O2诱导的胰岛细胞凋亡,增加细胞和组织抗氧化酶SOD的活性,PI3K-Akt抑制剂可阻断SFPS抗氧化作用,但不能最终抑制SFPS对胰岛细胞数量和活性的保护作用,说明SFPS对胰岛细胞的保护作用可能还存在除PI3K-Akt通路以外的其他通路。
Objective: This experiment was designed to understand the protective effect and mechanism of Sargassum Fusiforme Polysaccharides (SFPS) on islet β cells. Methods: The apoptosis of rat islet cell line (RINm5F) was induced by H2O2 in vitro. The effect of SFPS on the activity of RINm5F was detected by MTT assay. The effect of SFPS on SOD and MDA of RINm5F and primary rat islets were detected by spectrophotometry. RESULTS: Apoptosis of RINm5F was induced by 0.2mmol/L H2O2, typical apoptosis was observed in Hoechst3342/PI, and LY294002 and H2O2, a 30μmol/L PI3K inhibitor, had a synergistic effect on cell damage; 20μg.ml-1SFPS protected H2O2 from injury to RINm5F activity. Treatment for 1 h did not block its protective effect; SOD was decreased in RINm5F and primary rat islet cells in the model group; SFPS could increase SOD activity in RINm5F and primary rat islets; LY294002 pretreatment blocked SFPS from increasing antioxidant enzyme activity. . CONCLUSION: H2O2 can induce apoptosis of islet cells, decrease the activity of antioxidant enzymes in cells and tissues, and have a synergistic effect with inhibitors of PI3K-Akt pathway. SFPS can reduce the apoptosis of islet cells induced by H2O2 and increase the cell The activity of anti-oxidative enzymes SOD is organized, PI3K-Akt inhibitors can block the antioxidation of SFPS, but can not ultimately inhibit the protective effect of SFPS on islet cell number and activity, indicating that the protective effect of SFPS on islet cells may still exist in addition to PI3K- Other pathways beyond the Akt pathway.