盐酸青藤碱对db/db小鼠肾组织超微结构及干扰素基因刺激因子表达的影响  

Effects of sinomenine hydrochloride on ultrastructure and STING expression in kidney tissues of db/db mice

作　　者：贺彤彤 靳晓飞 左春月 周晓红 高钰 陈香美 高维娟 HE Tongtong;JIN Xiaofei;ZUO Chunyue;ZHOU Xiaohong;GAO Yu;CHEN Xiangmei;GAO Weijuan(Hebei University of Traditional Chinese Medicine,Hebei Key Laboratory of Chinese Medicine Research on Cardio-cerebro-vascular Disease,Shijiazhuang 050200,China;The First Medical Center of Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]河北中医药大学,河北省心脑血管病中医药防治研究重点实验室,河北石家庄050200 [2]中国人民解放军总医院第一医学中心,北京100853

出　　处：《中国病理生理杂志》2024年第11期2074-2080,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金专项资助项目(No.32141005);河北省研究生创新资助项目(No.CXZZSS2023087)。

摘　　要：目的:观察盐酸青藤碱对db/db小鼠肾组织超微结构和干扰素基因刺激因子(STING)表达的影响。方法:将16只12周龄雄性db/db小鼠随机分为2组:模型(model)组和盐酸青藤碱(SIN)组,每组8只,并设8只野生型(WT)小鼠8只小鼠为正常(control)组。盐酸青藤碱组连续灌胃给药8周后在20周末进行相关指标检测,正常组和模型组给予等体积生理盐水灌胃处理。每周检测各组小鼠体重和空腹血糖,全自动生化分析仪检测小鼠尿素氮(BUN)和血清肌酐(SCr)含量,酶联免疫吸附法(ELISA)检测24 h尿微量白蛋白(ALB)和血清中炎症因子白细胞介素1β(IL-1β)、IL-6和肿瘤坏死因子α(TNF-α)含量;HE染色观察各组小鼠肾组织的病理变化;透射电子显微镜观察各组小鼠肾组织的超微结构变化;免疫组化法和Western blot法检测各组小鼠肾组织STING蛋白以及核因子NF-κB的亚基P65蛋白的表达;RT-qPCR检测各组小鼠肾组织STING的mRNA表达。结果:与正常组相比,模型组小鼠肾功能指标BUN、ALB和SCr含量均显著上升(P<0.01),炎症因子IL-1β、IL-6和TNF-α含量均显著上升(P<0.01);毛细血管内可见白细胞附壁,炎症细胞浸润,系膜外基质增多;足突融合明显呈线状且排列紊乱,基底膜增厚;STING蛋白和mRNA以及P65蛋白表达水平均显著升高(P<0.01)。与模型组相比,盐酸青藤碱组小鼠肾功能指标BUN、SCr和ALB含量均显著下降(P<0.01),炎症因子IL-1β、IL-6和TNF-α含量均显著下降(P<0.01);毛细血管内白细胞附壁减少,炎症细胞和系膜外基质减少;足突融合现象显著减轻,基底膜厚度变薄;STING蛋白和mRNA以及p65蛋白表达水平均显著减少(P<0.01)。结论:盐酸青藤碱可以减轻db/db小鼠肾组织损伤,改善肾组织超微结构变化,抑制炎症反应,其作用机制与下调STING蛋白和mRNA表达紧密联系。AIM:This study aimed to investigate the effects of sinomenine hydrochloride(SIN)on the ultra-structure of renal tissue and the expression of interferon gene-stimulating factor in db/db mice.METHODS:Sixteen 12-week-old male db/db mice were randomly divided into two groups:a model group and a sinomenine hydrochloride(SIN)group,each consisting of 8 mice.An additional 8 wild-type(WT)mice served as the normal control group.The sinome-nine hydrochloride group was administered the treatment for 8 weeks,followed by a 20-week observation period,while the normal and model groups received an equal volume of saline via gavage.Weekly measurements were taken for body weight and fasting blood glucose.Serum creatinine(SCr)and blood urea nitrogen(BUN)levels were assessed,and 24-hour uri-nary microalbumin(ALB)levels,as well as serum inflammatory cytokines interleukin-1β(IL-1β),IL-6 and tumor necro-sis factor-α(TNF-α),were determined using ELISA.Pathological changes in renal tissue were evaluated through hema-toxylin-eosin(HE)staining,while ultrastructural alterations were examined using transmission electron microscopy.Im-munohistochemistry and Western blotting were employed to assess STING protein expression in renal tissue,and STING mRNA expression was quantified via RT-qPCR.RESULTS:Compared to the normal group,the model group exhibited significant increases in BUN,ALB,and SCr levels(P<0.01),alongside elevated inflammatory markers IL-1β,IL-6,and TNF-α(P<0.01).Notable pathological changes included leukocyte wall thickening in capillaries,inflammatory cell infiltration,increased mesangial matrix,disorganized and linear alignment of podocytes,and thickening of the basement membrane.Moreover,STING protein and mRNA expression levels were significantly elevated(P<0.01).In contrast,the sinomenine hydrochloride group demonstrated significantly reduced levels of renal function markers(BUN,ALB and SCr)compared to the model group(P<0.01),as well as decreased concentrations of inflammatory factors IL-1β,IL-6,and TNF-α(P<0.01).Improv

关 键 词：糖尿病肾病 盐酸青藤碱 炎症 干扰素基因刺激因子 超微结构 

分 类 号：R587.2[医药卫生—内分泌] R285.5[医药卫生—内科学] R363[医药卫生—临床医学]