D-柠檬烯对衣霉素诱导的胰腺MIN6细胞损伤的保护作用

D-limonene Protects Pancreatic MIN6 Cells against Tunicamycin Induced Cellular Damage

本研究主要探讨D-柠檬烯对衣霉素(Tm)诱导胰腺MIN6细胞损伤的保护作用。设置溶剂对照组、Tm组、4-苯基丁酸(PBA)组和不同浓度的D-柠檬烯组,用Griess试剂检测Tm刺激胰岛β细胞产生的NO水平;Annexin V-FITC/PI双染法流式检测细胞凋亡; RT-PCR法检测mRNA的表达水平; Western Blot法检测相关蛋白的表达。结果显示,D-柠檬烯组与Tm组相比,NO产生量减少、细胞凋亡率降低、内质网应激相关基因sXbp1和CHOP的mRNA表达量显著降低、p-eIF2α的蛋白表达量显著下降。结果表明D-柠檬烯对衣霉素(Tm)诱导的胰腺MIN6细胞损伤具有显著的保护作用。

In this study,the protective effect of D-limonene on the cellular damage induced by tunicamycin in pancreatic MIN6 cells was investigated.Cells were pretreated with vehicle,PBA and different doses of D-limonene,and then cultured with Tm for 5-24 h.NO production was detected by Griess reagent method.Cells were stained with Annexin V-FITC/PI for apoptosis by flow cytometry.RT-PCR was used to analyze mRNA expression of ER stress related genes sXbp1 and CHOP.Western Blot was used to detect eIF2αprotein expression.Our results showed that,compared with Tm-treated cells,cells treated with D-limonene exhibited significantly decreased NO production and apoptosis.Furthermore,D-limonene treatment resulted in significantly lower mRNA levers of sXbp1 and CHOP as well as lower level of protein expression of phosphorylated eIF2α.These results demonstrated that D-limonene had a significant protective effect on cellular damage induced by tunicamycin in pancreatic MIN6 cells.