续苓健骨方联合阿仑膦酸钠调控EphB4/EphrinB2双向通路对去卵巢骨质疏松大鼠的研究

Regulation of Xuling-Jiangu formula combined with alendronate on EphB4/EphrinB2 signaling pathway in ovariectomized rats

目的观察续苓健骨方联合阿仑膦酸钠对去卵巢快速骨丢失模型大鼠骨密度、结构、骨代谢和EphB4/EphrinB2信号通路的影响,探讨其作用机制。方法50只雌性SD大鼠随机分为5组:假手术组、模型组、续苓健骨方组、阿仑膦酸钠组和联合用药组,行双侧卵巢切除术制备模型;药物分别干预12周后取材,DXA检测股骨骨密度,HE染色观察胫骨组织显微结构,ELISA检测血清Ⅰ型胶原交联C末端肽(S-CTX)和Ⅰ型原胶原N端前肽(PINP)水平,Real-time PCR和Western blot检测腰椎EphB4、EphrinB2 mRNA和蛋白的表达水平。结果与假手术组相比,模型组骨密度显著降低(P<0.001),骨小梁稀疏并且排列紊乱,S-CTX和PINP含量均增高(P<0.01),骨组织EphB4、EphrinB2 mRNA和蛋白表达水平均显著降低(P<0.001)。与模型组相比,3个药物组骨密度均增高,其中以联合用药组最佳(P<0.001),骨小梁较密并且排列规则,S-CTX含量不同程度降低,PINP含量不同程度增高,EphB4、EphrinB2 mRNA和蛋白表达水平均增高。结论续苓健骨方联合阿仑膦酸钠起协同作用,可能通过激活EphB4/EphrinB2信号通路改善骨组织显微结构和血清骨代谢指标,从而提高大鼠骨密度起抗骨质疏松作用。

Objective To observe the effects of Xuling-Jiangu formula combined with alendronate on bone mineral density(BMD),structure,bone metabolism,and EphB4/ephrinB2 signal pathway in ovariectomized rats with rapid bone loss,and to explore its mechanism.Methods Fifty female SD rats were randomly divided into 5 groups:sham operation group,model group,Xuling-Jiangu formula group,alendronate group,and combined treatment group.Postmenopausal osteoporosis model was established with bilateral ovariectomy.After 12 weeks of drug intervention,BMD of the femur was measured using dual energy X-ray absorptiometry.The microstructure of the tibia was observed with HE staining.The levels of serum C-terminal cross linked peptide(S-CTX)and type I procollagen N-terminal propeptide(PINP)were detected with ELISA.The expression levels of EphB4,ephrinB2 mRNA and protein in the lumbar spine were detected with real-time PCR and Western blotting.Results Compared to those in the sham operation group,BMD in the model group decreased significantly(P<0.001),the bone trabeculae were sparse and disordered,and the contents of S-CTX and PINP increased(P<0.01).The expression levels of EphB4,ephrinB2 mRNA and protein in bone tissue decreased significantly(P<0.001).Compared to that in the model group,BMD in the three drug groups increased,and it was the best in the combined drug group(P<0.001).The trabeculae were dense and arranged regularly,the content of S-CTX decreased in varying degrees,and the content of PINP increased in varying degrees.The expression levels of EphB4,ephrinB2 mRNA and protein increased.Conclusion Xuling-Jiangu formula combined with alendronate may improve bone microstructure and serum bone metabolism by activating EphB4/ephrinB2 signal pathway,so as to improve BMD and to play an anti-osteoporosis role in rats.