三黄糖肾康对糖尿病大鼠骨组织Wnt/β-catenin信号通路的影响  

Effect of Sanhuang Tangshenkang on Wnt/β-catenin Signaling Pathway in Bone Tissue of Diabetic Rats

作　　者：孙立亚 谷丽艳[1,2] 刘北 王嘉熙 冯一楠 奚悦 SUN Liya;GU Liyan;LIU Bei;WANG Jiaxi;FENG Yinan;XI Yue(Liaoning University of Traditional Chinese Medicine(TCM),Shenyang 110847,China;Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,Liaoning University of TCM,Shenyang 110847,China;Shenyang New District Health Supervision Center,Shenyang 110127,China;The Third Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,China)

机构地区：[1]辽宁中医药大学,沈阳110847 [2]辽宁中医药大学中医脏象理论及应用教育部重点实验室,沈阳110847 [3]沈北新区卫生健康监督中心,沈阳110127 [4]锦州医科大学附属第三医院,辽宁锦州121001

出　　处：《中国实验方剂学杂志》2023年第18期69-77,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：辽宁省应用基础研究计划项目(2022JH2/101300059)。

摘　　要：目的:观察三黄糖肾康对糖尿病大鼠骨组织Wnt/β-连环蛋白(β-catenin)信号通路的影响。方法:采用高糖高脂饮食4周联合左下腹腔内注射新鲜配制的2%链脲佐菌素(pH 4.5)30 mg·kg^(-1)体质量制备糖尿病模型,成模大鼠按血糖水平随机分为模型组、三黄糖肾康低、高剂量组(12.8、38.4 g·kg^(-1))、骨疏康组(1.8 g·kg^(-1)),另设同周龄喂食普通饲料大鼠为空白组。各组予相应药物或生理盐水灌胃12周后,全自动生化仪检测各组大鼠空腹血糖(FBG);酶联免疫吸附测定法(ELISA)检测血清空腹胰岛素(FINS)、骨特异性碱性磷酸酶(BALP)、抗酒石酸酸性磷酸酶(TRAP);计算机微断层扫描(Micro-CT)扫描大鼠股骨,观察骨组织微结构,检测骨密度(BMD);苏木素-伊红(HE)染色、番红O-固绿染色观察大鼠股骨组织病理变化;免疫组化(IHC)和蛋白免疫印迹法(Western blot)检测Wnt3a、低密度脂蛋白受体相关蛋白5(LRP-5)及β-catenin蛋白的表达。结果:与空白组比较,模型组大鼠FBG、FINS、TRAP显著升高(P<0.01),BALP显著降低(P<0.01),BMD显著降低(P<0.01),骨小梁结构松散不规则,细长疏稀,出现断裂,脂滴增多,Wnt3a、LRP-5和β-catenin蛋白表达下降(P<0.05,P<0.01);与模型组比较,三黄糖肾康低、高剂量组FBG降低、BALP升高(P<0.05),三黄糖肾康低剂量组FINS降低(P<0.05),各给药组TRAP均显著降低(P<0.01);各给药组BMD均有不同程度提高(P<0.05,P<0.01);各给药组大鼠股骨骨小梁增多、增粗,排列较紧密、整齐,脂滴数量减少,骨微结构形态得到一定的改善;各给药组Wnt3a、LRP-5和β-catenin蛋白平均光密度值均有不同程度提高(P<0.05,P<0.01);Wnt3a、LRP-5和β-catenin蛋白表达升高(P<0.05,P<0.01)。结论:三黄糖肾康可能通过提高骨组织Wnt3a、LRP-5及β-catenin蛋白的表达,调节Wnt/β-catenin信号传导通路的失调状态,促进骨形成,减少骨吸收,降低血糖,从而达到防治糖尿病骨质疏松症的效果。Objective:To observe the effect of Sanhuang Tangshenkang on the Wnt/β-catenin signaling pathway in the bone tissue of diabetic rats.Method:A high-sugar and high-fat diet was administered for 4 weeks,along with intraperitoneal injection of freshly prepared 2%streptozotocin(pH 4.5)at 30 mg·kg^(-1) body weight to induce a diabetes model in rats.The rats with diabetes were randomly divided into model group,lowand high-dose Sanhuang Tangshenkang groups(12.8,38.4 g·kg^(-1)),and Gushukang group(1.8 g·kg^(-1))according to the blood glucose level.Rats of the same age were fed on a regular diet and assigned to the control group.After 12 weeks of respective treatments with drugs or physiological saline,the fasting blood glucose(FBG)levels of the rats were measured using an automated biochemical analyzer.Enzyme-linked immunosorbent assay(ELISA)was used to detect fasting serum insulin(FINS),bone-specific alkaline phosphatase(BALP),and tartrate-resistant acid phosphatase(TRAP)levels.Micro-computed tomography(Micro-CT)was used to scan the femurs of rats to observe bone tissue microstructure and measure bone mineral density(BMD).Hematoxylin-eosin(HE)staining and safranin O/fast green staining were performed to observe pathological changes in the femoral bone tissue.Immunohistochemistry(IHC)and Western blot were used to detect the expression of Wnt3a,low-density lipoprotein receptor-related protein 5(LRP-5),andβ-catenin proteins.Result:Compared with the control group,the model group showed a significant increase in FBG,FINS,and TRAP levels(P<0.01),a significant decrease in BALP level(P<0.01),a significant decrease in BMD(P<0.01),and disorganized,elongated,and sparse bone trabecular structures with fractures and increased lipid droplets.Additionally,the expression of Wnt3a,LRP-5,andβ-catenin proteins decreased(P<0.05,P<0.01).Compared with the model group,the low-and high-dose Sanhuang Tangshenkang groups showed a reduction in FBG and an increase in BALP(P<0.05).The low-dose Sanhuang Tangshenkang group also exhibited a decrea

关 键 词：糖尿病 骨质疏松症 三黄糖肾康 Wnt/β-连环蛋白 信号通路 

分 类 号：R2-0[医药卫生—中医学] R33R587.1R289R692