黄芪桂枝五物汤加减对糖尿病大鼠坐骨神经内质网应激IRE1α/CHOP通路的影响  

Effect of Modified Huangqi Guizhi Wuwutang on Stress-related IRE1α/CHOP Pathway of Sciatic Nerve Endoplasmic Reticulum in Diabetes Rats

作　　者：张岩[1] 龙泓竹[2] 王曦鹏 曹伟芳 吴丽丽 刘铜华 周静鑫[6] ZHANG Yan;LONG Hongzhu;WANG Xipeng;CAO Weifang;WU Lili;LIU Tonghua;ZHOU Jingxin(Beijing Friendship Hospital Affiliated to Capital Medical University,Beijing 100050,China;China-Japan Friendship Hospital,Beijing 100029,China;Hospital of Beijing International Studies University,Beijing 100024,China;Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences,Beijing 100007,China;School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]首都医科大学附属北京友谊医院,北京100050 [2]中日友好医院,北京100029 [3]北京第二外国语学院校医院,北京100024 [4]中国医学科学院基础医学研究所,北京100007 [5]北京中医药大学中医学院,北京100029 [6]北京中医药大学东直门医院,北京100700

出　　处：《中国实验方剂学杂志》2023年第16期43-51,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81804091)。

摘　　要：目的:基于肌醇需求酶1α(IRE1α)和CCAAT/增强子结合蛋白同源蛋白(CHOP)通路,研究黄芪桂枝五物汤加减对糖尿病大鼠坐骨神经在内质网应激方面的保护作用。方法:取60只大鼠予高糖高脂饲料喂养6周,然后按35 mg·kg^(-1)腹腔注射链尿佐菌素,3 d后检测大鼠随机血糖,连续检测3 d,将血糖持续≥16.7 mmol·L^(-1)的大鼠纳入实验,共48只。将48只大鼠随机分为模型组、α-硫辛酸组(0.0268 g·kg^(-1)·d^(-1))、中药高、低剂量组(2.5、1.25 g·kg^(-1)·d^(-1)),另设置10只为正常组。连续干预16周。16周后通过卢卡斯快蓝染色(LFB)光镜下观察各组大鼠坐骨神经结构,透射电镜观察各组大鼠坐骨神经超微结构,采用化学荧光法检测大鼠血清活性氧(ROS)含量,蛋白免疫印迹法和实时荧光定量聚合酶链式反应分别检测大鼠坐骨神经磷酸化IRE1α(p-IRE1α)蛋白、IRE1αmRNA、CHOP蛋白和mRNA的表达。结果:与正常组比较,模型组大鼠血清中ROS含量均显著升高(P<0.01);与模型组比较,各给药组大鼠血清中ROS含量显著降低(P<0.01)。与正常组比较,模型组大鼠坐骨神经出现病理改变;与模型组比较,各给药组大鼠坐骨神经病理有所改善。与正常组比较,模型组大鼠坐骨神经(p-IRE1α)、CHOP蛋白表达显著升高(P<0.01);与模型组比较,各给药组大鼠坐骨神经p-IRE1α、CHOP蛋白表达明显降低(P<0.05,P<0.01)。与正常组比较,模型组大鼠坐骨神经IRE1α、CHOP mRNA表达显著升高(P<0.01);与模型组比较,各给药组大鼠坐骨神经IRE1α和CHOP mRNA表达显著降低(P<0.01)。结论:黄芪桂枝五物汤加减可以通过抑制内质网应激IRE1α/CHOP途径相关蛋白和mRNA的表达,从而减轻内质网应激诱导的细胞凋亡,改善糖尿病大鼠坐骨神经结构和功能,从而防治糖尿病周围神经病变。Objective:To investigate the protective effect of modified Huangqi Guizhi Wuwutang(MHGW)on endoplasmic reticulum stress in the sciatic nerve of diabetes rats based on the pathways of inositolrequiring enzyme 1α(IRE1α)and CCAAT/enhancer-binding protein homologous protein(CHOP).Method:Sixty rats were fed on a high-sugar and high-fat diet for six weeks,followed by intraperitoneal injection of streptozotocin at a dose of 35 mg·kg^(-1).Random blood glucose levels were measured three days later and rats with a sustained blood glucose level≥16.7 mmol·L^(-1) were included in study(n=48).The rats were randomly divided into a model group,anα-lipoic acid group(0.0268 g·kg^(-1)·d^(-1)),a high-dose MHGW group(2.5 g·kg^(-1)·d^(-1)),and a low-dose MHGW group(1.25 g·kg^(-1)·d^(-1)).Another 10 rats were assigned to the normal group.The intervention lasted for 16 weeks.After 16 weeks,the sciatic nerve structure of the rats in each group was observed under light microscopy using Luxol fast blue(LFB)staining.Transmission electron microscopy was used to observe the ultrastructure of the sciatic nerve.Chemiluminescence method was employed to measure the serum reactive oxygen species(ROS)levels.Western blot and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)were used to evaluate the expression of p-IRE1αprotein,IRE1αmRNA,CHOP protein,and CHOP mRNA in the sciatic nerve of the rats.Result:Compared with the normal group,the model group showed elevated serum ROS levels(P<0.01).In contrast,the serum ROS levels were significantly reduced in the treatment groups compared with those in the model group(P<0.01).The sciatic nerve of the model group showed pathological changes compared with that in the normal group,while the treatment groups exhibited improvement in sciatic nerve pathology compared with the model group.The protein expression of p-IRE1αand CHOP in the sciatic nerve significantly increased in the model group as compared with that in the normal group(P<0.01).However,the treatment groups s

关 键 词：黄芪桂枝五物汤加减 糖尿病周围神经病变 肌醇需求酶1α(IRE1α) 磷酸化IER1α(p-IRE1α) CCAAT/增强子结合蛋白同源蛋白 

分 类 号：R2-0[医药卫生—中医学] R22R242R2-031R285.5