摘要
目的 基于Wnt/β-联蛋白(β-catenin)信号通路探讨坚骨胶囊对骨质疏松症模型大鼠的作用及机制。方法 将50只Wistar雌性大鼠随机分为正常对照组,模型对照组,坚骨胶囊高、中、低剂量组[2 184,1 092,546 mg/(kg·d)],各10只。除正常对照组外,其余各组大鼠均灌胃100 mg/kg维甲酸复制骨质疏松症模型,每天1次,连续30 d。坚骨胶囊高、中、低剂量组大鼠灌胃相应剂量药物,正常对照组和模型对照组大鼠均灌胃纯化水8 mL/kg,每天1次,连续30 d。检测各组大鼠血清的生化指标Ca2+、Mg2+、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、碱性磷酸酶(ALP)水平;利用Micro-CT仪记录骨微结构,并检测大鼠股骨微结构指标[远心端感兴趣区结构模型指数(SMI)、骨密度(BMD)、骨体积分数(BV/TV)、骨表面与骨体积比(BS/BV)、骨小梁数量(TbN)、骨小梁厚度(TbTh)、骨小梁分离度(TbSp)];采用免疫印迹(Western blot)法检测股骨组织中细胞周期蛋白D1(CyclinD1)、β-catenin、低密度脂蛋白受体相关蛋白5(LRP5)、淋巴增强因子(LEF)的蛋白表达水平。结果 MicroCT仪检测结果显示,坚骨胶囊高剂量组大鼠骨小梁结构最完整,与正常对照组股骨远心端骨小梁多呈板状、致密均匀接近。与模型对照组比较,坚骨胶囊中、高剂量组大鼠血清中MDA,GSH-Px,SOD水平均明显升高(P <0.05),ALP水平均明显降低(P <0.05);坚骨胶囊高剂量组大鼠股骨微结构中BMD,BV/TV,BS/BV,TbN,TbTh均明显升高,SMI和TbSp均明显降低(P <0.05);坚骨胶囊高剂量组大鼠股骨组织中LRP5,β-catenin,LEF,CyclinD1蛋白表达均明显上调(P <0.05)。结论 坚骨胶囊能预防维甲酸诱导的大鼠骨质疏松,其作用机制可能与激活Wnt/β-catenin信号通路及其基因蛋白表达有关。
Objective To investigate the effect and mechanism of Jiangu Capsules on the osteoporosis model rats based on the Wnt/β-catenin signaling pathway.Methods A total of 50 female Wistar rats were randomly divided into the normal control group,the model control group,the Jiangu Capsules high-,medium-and low-dose groups[2184,1092,546 mg/(kg•d)],with 10 rats in each group.Except for the normal control group,the rats in the other groups were given 100 mg/kg of tretinoin by gavage to replicate the osteoporosis model,once a day,for 30 d.The rats in the Jiangu Capsules high-,medium-and low-dose groups were given corresponding doses of drugs by gavage,and the rats in the normal control group and the model control group were given 8 mL/kg of purified water by gavage,once a day,for 30 d.The levels of serum biochemical indexes Ca2+,Mg2+,malondialdehyde(MDA),glutathione peroxidase(GSH-Px),total antioxidant capacity(T-AOC),superoxide dismutase(SOD)and alkaline phosphatase(ALP)in each group were detected.The Micro-CT was used to record the femoral microstructure and detect the related indexes[structural model index(SMI),bone mineral density(BMD),bone volume/total volume(BV/TV),bone surface/bone volume(BS/BV),trabecular number(TbN),trabecular thickness(TbTh),trabecular separation(TbSp)at distal region of interest].The Western blot was used to detect the expression levels of CyclinD1,β-catenin,low-density lipoprotein receptor-related protein 5(LRP5)and lymphoid enhancing factor(LEF).Results The results of Micro-CT showed that the trabecular structure was the most complete,and the trabecular at the distal end of the femur in the Jiangu Capsules high-dose group was mostly plate-like,dense and uniform,which was similar to that in the normal control group.Compared with those in the model control group,the levels of MDA,GSH-Px and SOD in serum of rats in the Jiangu Capsules medium-and high-dose groups were significantly higher(P<0.05),the ALP level in the Jiangu Capsules medium-and high-dose groups was significantly lower(P<0.05).Compared with those in the model control group,BMD,BV/TV,BS/BV,TbN and TbTh in the femoral microstructure of rats in the Jiangu Capsules high-dose group were significantly higher,while SMI and TbSp in the Jiangu Capsules high - dose group were significantly lower (P < 0. 05). Compared with those in the model control group,the expressionlevels of LRP5,β - catenin,LEF and CyclinD1 proteins in femoral tissue of rats in the Jiangu Capsules high - dose group weresignificantly higher (P < 0. 05). Conclusion Jiangu Capsules can prevent the osteoporosis of rats induced by tretinoin,and itsmechanism may be related to the activation of Wnt / β - catenin signaling pathway and the expression of related proteins.
作者
金颖慧
闫国强
张倩
张俊艳
董晶晶
JIN Yinghui;YAN Guoqiang;ZHANG Qian;ZHANG Junyan;DONG Jingjing(Hebei Cangzhou Hosptial of Integrated TCM-WM,Cangzhou,Hebei,China 061000)
出处
《中国药业》
CAS
2023年第7期56-60,共5页
China Pharmaceuticals
基金
河北省中医药管理局[2020518]。
