阿格列汀对卵巢切除小鼠骨量丢失的影响

Effect of alogliptin on bone loss in ovariectomized mice

目的探讨阿格列汀对卵巢切除小鼠骨量丢失的影响。方法(1)动物实验:取30只8周龄雌性C57BL/6J小鼠,将其分为对照组(Sham组)、卵巢切除组(ovariectomized,OVX组)、卵巢切除加阿格列汀干预组(OVX+Alogliptin组),每组10只。实验组以20 mg·kg^(-1)·d^(-1)阿格列汀进行灌胃干预,对照组给予等量生理盐水。连续干预12周后取材,检测其血清骨合成代谢相关指标,分别利用Micro CT扫描及HE染色观察小鼠股骨和胫骨的骨小梁结构并进行形态学分析。(2)细胞实验:取P3代骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs),以100μmol/L阿格列汀进行成骨诱导培养后,进行碱性磷酸酶(ALP)和茜素红S染色,测定细胞ALP活性和矿化能力;采用实时荧光定量PCR和Western印迹法检测成骨相关基因mRNA及蛋白表达水平。结果阿格列汀干预可提高卵巢切除小鼠骨合成代谢生化指标,改善卵巢切除小鼠骨微结构破坏,缓解骨量丢失。在体外实验中,阿格列汀促进BMSCs成骨分化,提高成骨基因Runt相关转录因子2(Runx2)、ALP、成骨相关转录因子(osterix)及骨钙素(osteocalcin)表达水平。结论阿格列汀可延缓卵巢切除小鼠的骨量丢失。

Objective To investigate the effect of alogliptin on bone loss in ovariectomized(OVX)mice.Methods For animal experiments,thirty 8-week-old C57BL/6J female mice were divided into Sham group,OVX group,and OVX+alogliptin group.OVX+alogliptin group were administered with alogliptin in a dosage of 20 mg·kg-1·d-1 by gavage,Sham and OVX groups with equivalent saline.After 12 weeks intervention,serum bone anabolism indicators were detected,and Micro CT and HE staining were used to observe and analyze the bone trabecular structure of femur and tibia in mice.For in vitro experiments,bone marrow mesenchymal stem cells(BMSCs)were incubated with 100μmol/L alogliptin for osteoblast differentiation.Alkaline phosphatase(ALP)and alizarin red S staining were used to determine the ALP activity and mineralization after osteogenic induction and culture.Real-time fluorescence quantitative PCR and Western blot were used to detect mRNA and protein expressions of osteoblast related genes.Results Alogliptin intervention improved the biochemical indexes of bone anabolism and protected against bone microstructure deterioration to alleviate bone loss in OVX mice.Alogliptin stimulated osteoblast differentiation and elevated expression levels of Runt-related transcription factor 2(Runx2),ALP,osteocalcin,and osterix in in vitro experiments.Conclusion Alogliptin can alleviate bone loss in OVX mice.