补肾健脾法调控自噬相关PINK1/Parkin信号通路治疗糖尿病性骨质疏松症  

Treatment of Diabetes Osteoporosis by Regulating Autophagy Related PINK1/Parkin Signaling Pathway with the Method of Kidney-Tonifying and Spleen-Invigorating

作　　者：文晓晨 刘若实[2] 朱克[3] 刘子英 马晓燕[4] 宫成军[4] 修静 杨芳[5] WEN Xiaochen;LIU Ruoshi;ZHU Ke;LIU Ziying;MA Xiaoyan;GONG Chengjun;XIU Jing;YANG Fang(College of Integrated Traditional Chinese and Western Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;The First Affiliated Hospital of China Medical University,Shenyang 110001,China;The Fourth Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110101,China;The Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China;College of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]辽宁中医药大学中西医结合学院,沈阳110847 [2]中国医科大学附属第一医院,沈阳110001 [3]辽宁中医药大学附属第四医院,沈阳110101 [4]辽宁中医药大学附属医院,沈阳110032 [5]辽宁中医药大学中医学院,沈阳110847

出　　处：《中国中医基础医学杂志》2025年第4期649-654,共6页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家中医药管理局马晓燕全国名老中医药专家传承工作室建设项目(国中医药人教函〔2022〕75号);国家自然科学基金面上项目(81973719);辽宁省应用基础研究计划(2022JH2/101300105);辽宁省科技厅联合基金博士科研启动项目(2023-BSBA-220);辽宁中医药大学博士后启动项目(21601A2120)。

摘　　要：目的观察补肾健脾法对糖尿病性骨质疏松症(diabetic osteoporosis,DOP)小鼠骨组织磷酸酶及张力蛋白同源物诱导激酶1(PINK1)/帕金蛋白(Parkin)通路介导自噬的影响,探索补肾健脾法治疗DOP的机制。方法将60只C57BL/6小鼠随机分为正常组、模型组、阳性对照组、补肾阴组、补肾阳组、健脾组及补肾健脾组。采用高脂高糖饲料联合链脲佐菌素(STZ)腹腔注射建立DOP模型。连续灌胃干预12周后,检测空腹血糖(FPG)、糖化血红蛋白(HbA1c)、碱性磷酸酶(ALP)、骨钙素(BGP)、抗酒石酸酸性磷酸酶(TRAP)及钙磷代谢指标;通过HE染色观察股骨组织病理改变,Micro-CT分析骨微结构参数;Western blot检测骨组织PINK1、Parkin、微管相关蛋白1轻链3(LC3)、自噬接头蛋白(p62)的表达。结果与正常组相比,模型组FPG、HbA1c、TRAP显著升高(P<0.01),ALP、BGP显著降低(P<0.01),骨小梁骨密度(BMD)及骨体积分数(BV/TV)下降(P<0.01)。与模型组比较,各治疗组HbA1c显著降低(P<0.01),骨组织病理损伤及骨微结构改善。补肾健脾组ALP、BGP水平升高(P<0.01),TRAP水平下降(P<0.01),提示该组具有改善骨代谢的作用;补肾健脾组PINK1、Parkin及LC3-II/I表达水平增加(P<0.05),p62蛋白表达减少(P<0.01),提示线粒体自噬被激活。结论补肾健脾法通过激活PINK1/Parkin信号通路增强线粒体自噬,从而改善DOP骨代谢紊乱及骨微结构损伤。Objective To investigate the therapeutic mechanism of kidney-tonifying and spleen-invigorating therapy on diabetic osteoporosis(DOP)by targeting the PINK1/Parkin pathway-mediated mitochondrial autophagy.Methods Sixty C57BL/6 mice were randomized into seven groups:normal control,DOP model,positive control,kidney-Yin tonification,kidney-Yang tonification,spleen-invigorating,and kidney-tonifying and spleen-invigorating groups.A DOP model was established via a high-fat/high-sucrose diet combined with intraperitoneal streptozotocin(STZ)injections.After 12 weeks of intervention,fasting plasma glucose(FPG),glycosylated hemoglobin(HbA1c),alkaline phosphatase(ALP),osteocalcin(BGP),tartrate-resistant acid phosphatase(TRAP),calcium(Ca),and phosphorus(P)levels were measured.Femoral histopathology was analyzed by hematoxylin-eosin(HE)staining,bone microstructure parameters were quantified using micro-computed tomography(micro-CT),and protein expression of PINK1,Parkin,LC3-II/I,and p62 was assessed via Western blot.Results Compared to the normal group,the model group exhibited significantly elevated FPG,HbA1c,and TRAP levels(P<0.01),reduced ALP and BGP(P<0.01),and decreased trabecular bone mineral density(BMD)and bone volume fraction(BV/TV)(P<0.05).All treatment groups showed improved glycemic control(HbA1c reduction,P<0.01),ameliorated bone histopathology,and restored bone microstructure compared to the model group.Compared with the model group,all treatment groups showed significant reductions in HbA1c levels(P<0.01)and improvements in bone histopathological damage and microstructure.In the kidney-tonifying and spleen-invigorating group,ALP and BGP levels were elevated(P<0.01),while TRAP levels decreased(P<0.01),indicating its potential role in ameliorating bone metabolism.Furthermore,the kidney-tonifying and spleen-invigorating group exhibited increased expression levels of PINK1,Parkin,and LC3-II/I(P<0.05),along with reduced p62 protein expression(P<0.01),suggesting activation of mitochondrial autophagy(mitophagy).Conclusi

关 键 词：糖尿病性骨质疏松症 补肾健脾法 PINK1/Parkin信号通路 线粒体自噬 

分 类 号：R259[医药卫生—中西医结合]