·糖尿病基础研究·

　　钟宇华 梁华晟

　　基金项目：广西科技厅自然科学基金资助项目(0991294);广西北海市科学研究与技术开发计划项目(200901059)

　　作者单位：536000北海，广西医科大学第九附属医院内分泌科

　　【摘要】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。

　　【关键词】间歇性高糖;高糖;INS-1 细胞;内质网应激;氯沙坦

　　doi:10.3969/j.issn.1006-6187.2012.02.017

　　The inhibitive effect of losartan on endoplasmic reticulum stress in INS-1 induced by high glucose and fluctuating high glucose ZHONG Yu-hua, LIANG Hua-sheng. Department of Endocrinology, the Ninth Affiliated Hospital, Guangxi Medical University, Guangxi Beihai 536000, China

　　【Abstract】ObjectiveTo study whether losartan can protect INS-1 from endoplasmic reticulum stress induced by high glucose and fluctuating high glucose. MethodsCell viability was detected by TTT in groups of control , normal glucose, high glucose, fluctuating high glucose, losartan, losartan plus high glucose, losartan plus fluctuating high glucose.The expression of GRP78, CHOP, Caspase-12 and p-JNK and apoptosis were measured by Western blot and Annexin Ⅴ, and ROS were detected by CM2H2DCFDA after 96h. ResultsCompared with control and normal glucose group, the groups of high glucose and fluctuating high glucose decreased cell viability (all P<0.01), increased cell apoptosis and ROS(all P<0.01)，up-regulated the expression of GRP78, CHOP, Caspase-12 and p-JNK(all P<0.01). Cell viability was lower in fluctuating high glucose group than in high glucose group(all P<0.01), but cell apoptosis(P<0.01), the expression of GRP78(P<0.05), caspase-12(P<0.01) and p-JNK(P<0.01), level of ROS(P<0.01)were higher in fluctuating high glucose group than in high glucose group. However losartan inhibited cell apoptosis and production of ROS in high glucose or fluctuating high glucose group(all P<0.01) and inhibited the up-regulation of the expression of GRP78, CHOP, caspase-12 and p-JNK(all P<0.01). ConclusionsLosartan may protect INS-1 from endoplasmic reticulum stress induced by high glucose and fluctuating high glucose.

　　【Key words】Fluctuating high glucose; High glucose; INS-1 cell; Endoplasmic reticulum stress; Losartan