目的:研究黄芪注射液与葛根素注射液合用对小鼠糖尿病肾病内质网应激中蛋白激酶R样内质网激酶(PERK)通路相关因子的影响。方法:将12周龄的雄性KKAy小鼠随机分成模型组和治疗组,每组14只,分别注射生理盐水或黄芪、葛根素注射液合剂,14只同龄雄性C57BL/6J小鼠为正常组。于16周龄时处死小鼠,电镜观察各组小鼠肾脏形态改变,通过real-time PCR和Western blot法检测PERK、真核起始因子2α(e IF2α)和葡萄糖调节蛋白78(GRP78)的mRNA及蛋白表达。结果:模型组及治疗组小鼠肾小管上皮细胞细胞核、内质网和线粒体肿胀。模型组PERK、e IF2α及GRP78的mRNA及蛋白表达均高于正常组(P〈0.05),治疗组较模型组下降。结论:黄芪注射液同葛根素注射液联用可能通过减少PERK、e IF2α及GRP78的表达降低2型糖尿病KKAy小鼠肾脏过强的内质网应激。
AIM:To investigate the effects of astragalus injection combined with puerarin injection on endoplasmic reticulum stress through PERK pathway in diabetic nephropathy mice. METHODS:Male KKAymice were randomly divided into model group(injected with normal saline)and treatment group(injected with astragalus and puerarin).The male C57 BL /6J mice served as normal group. The mice were sacrificed 4 weeks after treatments for observing morphological changes under electron microscope. The renal tissues were collected to determine the expression of protein kinase Rlike endoplasmic reticulum kinase(PERK),eukaryotic initiation factor 2α(e IF2α)and glucose-regulated protein 78(GRP78)at mRNA and protein levels by real-time PCR and Western blot. RESULTS:Under electron microscope,the renal tubular epithelial cells in model group and treatment group showed the swelling of the nucleus,endoplasmic reticulum and mitochondria. The results of real-time PCR and Western blot showed that the expression of PERK,e IF2α and GRP78 at mRNA and protein levels in model group was higher than that in normal group(P 〈 0. 05),while that in treatment group was lower than that in model group. CONCLUSION:Astragalus injection combined with puerarin injection reduces the mRNA and protein expression of PERK,e IF2α and GRP78,thus inhibiting the endoplasmic reticulum stress in type 2 diabetic mice to protect the kidney function.