基于DAG/PKC信号通路探讨盐酸青藤碱对db/db小鼠肾脏血流动力学及微循环障碍的影响  

Sinomenine hydrochloride attenuates renal hemodynamics and microcircu⁃lation disturbance in db/db mice by inhibiting DAG/PKC signaling pathway

作　　者：刘均广 李猛 陈笑寒 冯智丽 周晓红 陈香美 高维娟 LIU Junguang;LI Meng;CHEN Xiaohan;FENG Zhili;ZHOU Xiaohong;CHENXiangmei;GAO Weijuan(Department of Pathophysiology,Chengde Medical College,Chengde 067000,China;Hebei University of Chinese Medicine,Hebei Key Laboratory of Chinese Medicine Research on Cardio-cerebrovascular Disease,Shijiazhuang 050091,China;Department of Nephrology,First Medical Center of Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]承德医学院病理生理学教研室,河北承德067000 [2]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050091 [3]中国人民解放军总医院第一医学中心肾脏病医学部,北京100853

出　　处：《中国病理生理杂志》2025年第1期72-80,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金专项项目(No.32141005)。

摘　　要：目的:基于二酰基甘油(DAG)/蛋白激酶C(PKC)信号通路,探讨盐酸青藤碱(SH)对2型糖尿病db/db小鼠肾脏血流动力学及微循环障碍的影响。方法:24只6周龄雄性db/db小鼠随机分为模型组(等体积生理盐水)、LY(1 mg·kg^(-1)·d^(-1) LY333531)组和SH(124.8 mg·kg^(-1)·d^(-1) SH)组,同时选取8只同周龄db/m小鼠设为对照组(等体积生理盐水),每日1次,定时灌胃给药,持续6周。检测各组小鼠体质量、空腹血糖(FBG)、尿微量白蛋白(mALB)、尿肌肝(UCr)和尿β2微球蛋白(β2-MG)水平;运用彩色多普勒超声测量小鼠右肾段动脉收缩期峰值流速(PSV)、舒张期末流速(EDV)和阻力指数(RI);采用ELISA法检测小鼠血清内皮素1(ET-1)、内皮型一氧化氮合酶(eNOS)、IV型胶原(Col IV)和层粘连蛋白(LN)水平,以及肾组织DAG和PKC含量;采用HE和PAS染色观察肾脏组织形态变化;透射电镜观察肾脏超微结构变化;分别采用免疫组化法和Western blot法检测小鼠肾组织PKC和p-PKC的表达。结果:与模型组相比,LY组和SH组小鼠体质量均出现不同程度下降(P<0.05),且LY组FBG显著降低(P<0.01);尿mALB和β2-MG水平显著降低(P<0.01),UCr水平显著升高(P<0.01);右肾段动脉PSV和EDV显著升高(P<0.01),且SH组RI显著降低(P<0.01);血清ET-1、Col IV和LN水平均显著降低(P<0.01),eNOS水平显著升高(P<0.01);肾组织DAG和PKC含量显著降低,p-PKC蛋白表达亦显著降低(P<0.05);肾组织病理改变和超微结构病理变化均有所改善。结论:盐酸青藤碱可通过抑制DAG/PKC信号通路减轻db/db小鼠肾损伤,其机制可能与改善肾脏血流动力学及微循环障碍有关。AIM:To explore the effect of sinomenine hydrochloride(SH)on microcirculation and renal function in type 2 diabetic db/db mice with a focus on the diacylglycerol(DAG)/protein kinase C(PKC)signaling pathway.METHODS:Eighteen 6-week-old male db/db mice were randomly divided into three groups:model group(saline),LY group(administered with 1 mg·kg^(-1)·d^(-1) LY333531),and SH group(administered with 124.8 mg·kg^(-1)·d^(-1) SH).Additionally,6 age-matched db/m mice comprised the control group(saline).Daily intragastric administration lasted 6 weeks.Body mass,fasting blood glucose(FBG),urinary microalbumin(mALB),urinary creatinine(UCr)and urinaryβ2-microglobulin(β2-MG)were measured in each group.The serum levels of endothelin-1(ET-1),endothelial nitric oxide synthase(eNOS),collagen type IV(Col IV)and laminin(LN),and the levels of DAG and PKC in renal tissues were analyzed by enzyme-linked immunosorbent assay(ELISA).The morphological changes of renal tissues were assessed using HE and PAS staining,and kidney ultrastructure was examined via transmission electron microscopy.Immunohistochemistry and Western blot were used to detect PKC and p-PKC levels in renal tissues.RESULTS:Compared with model group,the mice in both LY and SH groups showed decreased body mass(P<0.05),with significantly reduced FBG level in LY group(P<0.01).In addition,the urinary mALB andβ2-MG levels were markedly decreased(P<0.01),while UCr level was significantly increased(P<0.01).Serum ET-1,Col IV and LN levels were significantly lower(P<0.01),whereas eNOS level was notably higher(P<0.01).Renal tissue DAG and PKC levels,as well as p-PKC expression were significantly reduced(P<0.01).Improvements in renal tissue pathology and ultrastructure were observed.CONCLU⁃SION:Sinomenine hydrochloride improves microcirculation in diabetic db/db mice by modulating DAG/PKC signaling pathway,thus exerting protective effect on the kidney.

关 键 词：盐酸青藤碱 糖尿病肾病 血流动力学 微循环 DAG/PKC信号通路 

分 类 号：R285.5[医药卫生—中药学] R587.2[医药卫生—中医学] R363