青娥丸调控Wnt1/β-catenin信号通路治疗糖尿病性骨质疏松症

Treatment of diabetic osteoporosis with Qing’e pill by regulation through Wnt1/β-catenin signaling pathway

目的探究青娥丸对糖尿病性骨质疏松症(diabetic osteoporosis,DOP)小鼠模型Wnt/β-catenin通路的影响。方法采用高糖高脂饲料喂养联合腹腔注射链脲菌素建立DOP小鼠模型,对小鼠进行药物干预,分为空白组、模型组、青娥丸低、中、高剂量组及阿仑膦酸钠组,进行一般状态观察,股骨组织进行HE染色,验证造模成功情况。生化检测各组小鼠碱性磷酸酶(ALP)、尿素氮(BUN)、天冬氨酸转氨酶(AST)的水平,免疫组化检测股骨Wnt/β-catenin信号传导通路中相关蛋白Wnt1、β-catenin、Runx2表达水平,HE染色观察各组股骨组织细胞形态,qPCR检测股骨组织中GSK-3β、低密度脂蛋白受体相关基因5(LRP5)、COL1A1 mRNA水平。结果与对照组相比,模型组小鼠骨小梁间隔增大,骨小梁之间连接减少,小鼠多饮多尿少动,尾静脉血糖值增加(P<0.01),血清中ALP、BUN、AST含量和GSK-3βmRNA水平均增加(P<0.01),体重和LRP5、COL1A1 mRNA水平下调(P<0.01),Wnt1、β-catenin、Runx2蛋白呈少量阳性表达(P<0.01)。与模型组相比,青娥丸组和阿仑膦酸钠组骨小梁间隔变小,骨小梁间的连接增多,尾静脉血糖值下降(P<0.01),血清中ALP、BUN、AST含量和GSK-3βmRNA水平均减少(P<0.01),体重和LRP5、COL1A1 mRNA水平上调(P<0.01),Wnt1、β-catenin、Runx2蛋白阳性表达增多(P<0.01)。结论青娥丸可增强DOP小鼠骨组织的修复作用,其机制可能与Wnt1/β-catenin信号通路的激活有关。

Objective To investigate the effect of the Qing’e pill on the Wnt/β-catenin pathway in a mouse model of diabetic osteoporosis(DOP).Methods Mice were fed with high-sugar and high-fat diet combined with intra-abdominal injection of streptozotocin to establish a DOP mouse model.The mice were divided into blank group,model group,Qing’e pill low-dose group,Qing’e medium-dose group,Qing’e pill high-dose group,and alendronate group.The levels of alkaline phosphatase(ALP),urea nitrogen(BUN),and aspartate aminotransferase(AST)in each group were measured with biochemistry.The expression levels of Wnt1,β-catenin,and Runx2 in the femur were measured with immunohistochemistry.The expression levels of GSK-3β,low-density lipoprotein receptor-related protein 5(LRP5),and COL1A1 mRNA in the femoral tissues were measured with qPCR.Results Compared with mice in the control group,the model mice showed increased bone trabecular septum,decreased connections between bone trabeculae,more drink and less urination and move,increased tail vein glucose values(P<0.01),increased serum levels of ALP,BUN,AST,and GSK-3βmRNA(P<0.01),and down-regulated body weight and LRP5 and COL1A1 mRNA levels(P<0.01).Wnt1,β-catenin,and Runx2 proteins showed a small amount of positive expression(P<0.01).Compared with mice in the model group,mice in the Qing’e pill group and alendronate group showed smaller bone trabecular septa,increased inter-trabecular connections,decreased tail vein glucose values(P<0.01),decreased serum levels of ALP,BUN,AST,and GSK-3βmRNA(P<0.01),up-regulated body weight and LRP5,COL1A1 mRNA levels(P<0.01),Wnt1,andβ-catenin,and Runx2 protein positive expression was increased(P<0.01).Conclusion Qing’e pill enhances the repairing effect of bone tissue in DOP mice.The mechanism may be related to its activation of Wnt1/β-catenin signaling pathway.