黄芪三仙汤抗绝经后骨质疏松作用机制的研究  

Study on the Mechanism of Action on Huangqi Sanxian Decoction Against Postmenopausal Osteoporosis

作　　者：邓艳玲 翁文玉 招泽豪 施健新 陈灼均 邓煜 周志昆 DENG Yan-Ling;WENG Wen-Yu;ZHAO Ze-Hao;SHI Jian-Xin;CHEN Zhuo-Jun;DENG Yu;ZHOU Zhi-Kun(Guangdong Medical University,Dongguan 523808 Guangdong,China)

机构地区：[1]广东医科大学,广东东莞523808

出　　处：《广州中医药大学学报》2022年第4期898-907,共10页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81774344)。

摘　　要：【目的】观察黄芪三仙汤抗绝经后骨质疏松症(PMOP)的作用机制。【方法】体内研究:采用去势法建立绝经后骨质疏松大鼠模型。测量大鼠股骨骨密度,进行骨组织形态学观察及骨小梁形态计量学测定,采用蛋白免疫印迹(Western Blot)法检测骨组织磷酸化蛋白激酶B(p-Akt)1/蛋白激酶B(Akt)1、p-Akt2/Akt2、雌激素受体α(ERα)蛋白表达。体外研究:原代培养PMOP成骨细胞。分别采用细胞计数试剂盒8(CCK-8)、碱性磷酸酶染色法和茜素红染色法观察黄芪三仙汤血清对PMOP成骨细胞增殖、分化和矿化的影响,采用Western Blot法检测成骨细胞p-Akt1/Akt1、p-Akt2/Akt2、ERα及成骨分化相关标志物包括Runt相关转录因子2(Runx2)、成骨细胞特异性转录因子Osterix(Osx)、骨桥蛋白(OPN)等的蛋白表达。【结果】与模型组比较,黄芪三仙汤组大鼠骨小梁断裂、缺失情况均得到改善,骨密度升高,骨小梁数量增加、面积增大、分离度减小,骨组织p-Akt1/Akt1、p-Akt2/Akt2和ERα蛋白表达水平升高(P<0.05)。与正常血清组比较,黄芪三仙汤血清组PMOP成骨细胞数量增加,碱性磷酸酶活性增强,矿化结节增多,p-Akt1/Akt1、p-Akt2/Akt2和ERα以及Runx2、Osx、OPN蛋白表达水平升高(P<0.05),而加入PI3K/Akt抑制剂LY294002后,翻转了黄芪三仙汤血清对PMOP成骨细胞碱性磷酸酶活性及Runx2、Osx、OPN表达的促进作用。【结论】黄芪三仙汤通过抑制PI3K/Akt信号通路活化起到抗PMOP的作用。Objective To observe the mechanism of action of Huangqi Sanxian Decoction against postmenopausal osteoporosis(PMOP).Methods In vivo study:a rat model of PMOP was established by castration.The bone density of rat femur was measured,and bone histomorphology observation and bone trabecular morphometry were performed.The expression of phosphorylated protein kinase B(p-Akt)1/protein kinase B(Akt)1,p-Akt2/Akt2and estrogen receptorα(ERα)in bone tissues was detected by Western Blot.In vitro study:primary culture of PMOP osteoblasts was carried out.The effects of serum containing Huangqi Sanxian Decoction on proliferation,differentiation and mineralization of PMOP osteoblasts were observed by cell counting kit 8(CCK-8),alkaline phosphatase staining method and alizarin red staining method,respectively.The protein expression of pAKT1/Akt1,p-Akt2/Akt2,ERαand osteoblast differentiation-related markers including Runt-associated transcription factor 2(Runx2),osteoblast specific transcription factor Osterix(Osx),osteopontin(OPN)was detected by Western Blot.Results Compared with the model group,the fracture and loss of trabecular bone in Huangqi Sanxian Decoction group were improved,bone mineral density(BMD)was increased,the number and dimension of trabecular bone were increased,and the separation degree of trabecular bone was decreased,and the protein expression levels of p-Akt1/Akt1,p-Akt2/Akt2 and ERαin bone tissues were increased(P<0.05).Compared with the normal serum group,the number of PMOP osteoblasts was increased,alkaline phosphatase activity was increased,mineralized nodles were increased,and the expression levels of p-Akt1/Akt1,p-Akt2/Akt2 and ERαin PMOP osteoblasts as well as Runx2,Osx and OPN were increased in serum containing Huangqi Sanxian Decoction group(P<0.05).After the addition of PI3K/Akt inhibitor LY294002,the promotion effect of serum containing Huangqi Sanxian Decoction on alkaline phosphatase activity and Runx2,Osx and OPN expression in PMOP osteoblasts was reversed.Conclusion Huangqi Sanxian Decocti

关 键 词：黄芪三仙汤 绝经后骨质疏松 PI3K/AKT信号通路 成骨细胞 细胞培养 大鼠 

分 类 号：R285.5[医药卫生—中药学]