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目的观察不同糖脂浓度时内质网应激及miR-375表达的改变对大鼠胰岛RINm5F细胞增殖、凋亡的影响。方法RINm5F细胞分别培养在含不同糖脂浓度的培养液中,利用CCK-8法检测细胞的增殖活性,实时荧光定量PCR(qt-PCR)检测内质网应激相关基因XBP-1,CHOP以及miR-375的表达,Western blot检测凋亡基因caspase-3以及CHOP蛋白的表达。结果相同葡萄糖浓度时,随着棕榈酸浓度的增加,内质网应激相关基因XBP-1 mRNA、CHOP mRNA以及miR-375的表达水平增高(P<0.05),胰岛β细胞的增殖活性下降(P<0.05),Western blot检测可见CHOP及caspase-3蛋白表达均增加。结论在高糖高脂的作用下,内质网应激的诱导及miR-375的增加与胰岛β细胞的凋亡相关。
Objective To observe the effect of endoplasmic reticulum stress and miR-375 expression on the proliferation and apoptosis of rat pancreatic islet RINm5F cells with different concentrations of glycolipids. Methods RINm5F cells were cultured in culture medium containing different concentrations of glycolipids. Cell proliferation was detected by CCK-8 assay. The expression of stress-related genes XBP-1, CHOP and endoplasmic reticulum stress were detected by real-time quantitative PCR (qt- miR-375 expression, Western blot detection of apoptosis gene caspase-3 and CHOP protein expression. At the same glucose concentration, the expression of XBP-1 mRNA, CHOP mRNA and miR-375 in endoplasmic reticulum increased with the increase of palmitate concentration (P <0.05), and the proliferation activity of pancreatic β-cells decreased P <0.05). The expression of CHOP and caspase-3 protein increased with Western blot. Conclusion The induction of endoplasmic reticulum stress and the increase of miR-375 are associated with the apoptosis of pancreatic β-cells under the action of high glucose and high fat.