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目的观察丹酚酸B(Salvianolic acid B,Sal B)对间歇性高糖诱导的大鼠胰岛素瘤细胞(INS-1)c-Jun氨基末端激酶(JNK)活化和细胞凋亡的影响。方法 INS-1细胞与丹酚酸B预孵24 h后暴露于间歇性高糖(11.1 mmol·L~(-1)12 h,33.3 mmol·L~(-1)12 h)72 h。MTT法测定细胞活力,ELISA法测定葡萄糖刺激的胰岛素分泌能力,荧光探针DCFH-DA检测细胞内活性氧(ROS)水平,流式细胞术检测细胞凋亡水平,Western blot法检测胰十二指肠同源盒-1(PDX-1)蛋白表达和JNK活化水平。结果丹酚酸B可明显减轻间歇性高糖诱导的INS-1细胞损伤和凋亡(P<0.05,P<0.01),提高葡萄糖刺激的胰岛素分泌能力(P<0.05,P<0.01);与丹酚酸B预孵可明显降低细胞内ROS水平、抑制JNK活化,并提高PDX-1蛋白表达水平(P<0.05,P<0.01)。结论丹酚酸B可有效减轻间歇性高糖诱导的INS-1细胞损伤和凋亡,提高胰岛素分泌水平,其机制可能与降低细胞内ROS水平、抑制JNK活化和上调PDX-1蛋白表达有关。
Objective To investigate the effects of salvianolic acid B (Sal B) on activation and apoptosis of c-Jun N-terminal kinase (JNK) induced by intermittent high glucose in rat insulinoma cells (INS-1). Methods INS-1 cells and salvianolic acid B were exposed to intermittent high glucose (11.1 mmol·L -1 12 h, 33.3 mmol·L -1 12 h) for 72 h after being pre-incubated for 24 h. Cell viability was measured by MTT assay. Glucose-stimulated insulin secretion was measured by ELISA. Fluorescence probe DCFH-DA was used to detect intracellular reactive oxygen species (ROS) levels. Flow cytometry was used to detect apoptosis. Western blot was used to detect pancreaticoduodenal Intestinal homeobox 1 (PDX-1) protein expression and JNK activation levels. Results Salvianolic acid B could significantly reduce the injury and apoptosis of INS-1 cells induced by intermittent high glucose (P <0.05, P <0.01), and improve the glucose-stimulated insulin secretion ability (P <0.05, P <0.01) Salvianolic acid B preincubation could significantly reduce intracellular ROS levels, inhibit JNK activation and increase PDX-1 protein expression (P <0.05, P <0.01). Conclusion Salvianolic acid B can effectively reduce the injury and apoptosis induced by intermittent high glucose in INS-1 cells and increase the level of insulin secretion. The mechanism may be related to the decrease of intracellular ROS level, the inhibition of JNK activation and the up-regulation of PDX-1 protein expression.