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目的探讨氯沙坦对高糖及间歇性高糖诱导大鼠胰岛β细胞株INS-1细胞内质网应激的抑制作用。方法各种刺激作用96h后,MTT法检测空白(Con)组、正常葡萄糖(NG)组、高糖(HG)组、间歇性高糖(MG)组、氯沙坦(LT)组、氯沙坦高糖(LT+HG)组及氯沙坦间歇性高糖(LT+MG)组的细胞活力;免疫印迹检测内质网标记物糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、Caspase-12及磷酸化C-Jun氨基末端激酶(p-JNK)在INS-1细胞的表达;AnnexinⅤ检测细胞凋亡;细胞活性氧(ROS)含量应用CM2H2DCFDA试剂盒检测。结果与Con组及NG组相比,高糖及间歇性高糖抑制细胞活性、凋亡率升高(P均<0.01),同时上调GRP78、CHOP、Caspase-12及p-JNK表达(P均<0.01),ROS含量上升(P均<0.01)。MG组细胞活性低于HG组(P<0.01),凋亡率、ROS含量以及GRP78、Caspase-12、p-JNK的表达均高于HG组(P<0.05或P<0.01)。氯沙坦能抑制高糖及间歇性高糖诱导的细胞凋亡及ROS产生(P均<0.01),同时抑制GRP78、CHOP、Caspase-12及p-JNK的表达上调(P均<0.01)。结论氯沙坦可通过抑制高糖及间歇性高糖诱导INS-1细胞内质网应激作用从而减少细胞凋亡。
Objective To investigate the inhibitory effect of losartan on endoplasmic reticulum stress induced by high glucose and intermittent high glucose in rat pancreatic β-cell INS-1 cells. Methods After stimulating for 96 hours, MTT assay was used to detect the levels of IL-6 in the Con group, normal glucose (NG) group, high glucose group (HG), intermittent high glucose group (MG), losartan (LT) (LT + HG) group and losartan intermittent high glucose (LT + MG) group. The endoplasmic reticulum markers glucose regulatory protein 78 (GRP78) and C / EBP homologous protein CHOP), Caspase-12 and phosphorylated C-Jun N-terminal kinase (p-JNK) in INS-1 cells. AnnexinⅤ was used to detect the apoptosis of cells. The level of ROS was detected by CM2H2DCFDA kit. Results Compared with Con group and NG group, high glucose and intermittent high glucose inhibited cell activity and increased apoptosis (P <0.01), and up-regulated GRP78, CHOP, Caspase-12 and p-JNK <0.01), ROS content increased (P <0.01). The cell viability in MG group was lower than that in HG group (P <0.01). The apoptosis rate, ROS content and the expressions of GRP78, Caspase-12 and p-JNK in MG group were higher than those in HG group (P <0.05 or P <0.01). Losartan inhibited the apoptosis and ROS production induced by high glucose and intermittent high glucose (all P <0.01), and inhibited the expression of GRP78, CHOP, Caspase-12 and p-JNK (all P <0.01). Conclusion Losartan can reduce cell apoptosis by inhibiting the endoplasmic reticulum stress induced by high glucose and intermittent high glucose in INS-1 cells.