【摘要】 目的研究解毒通络调肝方对大鼠胰岛细胞瘤细胞(islet cell tumor cells,INS-1)糖毒性的保护作用机制。方法以大鼠胰岛细胞瘤细胞为研究对象,用MTT比色法检测解毒通络调肝方干预后INS-1细胞存活率的情况;不同浓度的解毒通络调肝方干预高糖诱导的INS-1细胞;通过葡萄糖刺激胰岛素分泌(glucose-stimulated insulin secretion test,GSIS)检验胰岛素分泌量;实时逆转录聚合酶链反应法测定INS1、INS2、胰十二指肠同源盒因子1(pancreatic and duodenal homeobox 1,PDX1)和MafA的mRNA表达水平;蛋白印迹实验检测INS-1细胞中内质网的跨膜蛋白[阻抗性激酶lα(inositol-requiring enzyme 1α,IRE1α)、双链RNA依赖性蛋白激酶样内质网激酶(protein kinase R-like endoplasmic reticulum kinase, PERK)和活化的转录因子6 (activating transcription factor 6,ATF6)]的表达。结果通过从TCMSP数据库中总结分析解毒通络调肝方中潜在成分,表明该方具有较好的稳定性和重复性;不同浓度的解毒通络调肝方对INS-1细胞活力无抑制作用(P> 0.05),但能够抑制高糖诱导的胰岛素分泌的降低(P<0.05),提高INS1、INS2、PDX1和MafA的mRNA表达(P<0.05),抑制IRE1α、PERK、ATF6蛋白表达(P<0.05)。结论解毒通络调肝方能够通过提高胰岛素合成相关基因的表达,促进INS-1细胞的胰岛素分泌,降低内质网应激,以恢复细胞内稳态平衡,减少糖毒性对β细胞损伤。更多还原

【Abstract】 Objective To study the protective effect of Jiedu Tongluo Tiaogan formula（JDTL）on glucose toxicity of INS-1 cells.Methods MTT method was used to observe the effect of JDTL on the survival rate of INS-1 cells; different doses of JDTL intervened INS-1 cells were induced by high glucose; glucose stimulated insulin secretion test and RT-PCR were used to determine the insulin secretion and the mRNA expression levels of INS1, INS2, PDX1 and MafA after drug intervention; the method of western blot was used to detect the expression levels of IRE1α, ATF6 and PERK.Results Different concentrations of JDTL had no effect on the survival rate of INS-1 cells. It could increase the secretion of insulin, the mRNA expression of INS1, INS2, PDX1 and MafA, and inhibit the expression of IRE1α, ATF6 and PERK.Conclusion JDTL can promote insulin release and reduce endoplasmic reticulum stress by increasing the expression of insulin synthesis related genes, so as to restore homeostasis of cells and reduce β-cell damage.更多还原