淫羊藿苷调节PTEN激活成骨基因表达缓解大鼠骨质疏松症的实验研究  

Experimental Study of Icariin Alleviating Osteoporosis by Regulating Expression of Osteogenic Genes Activated by PTEN in Rats

作　　者：金垚 徐鹏辉 卞泗善[2] 滕加文[2] JIN Yao;XU Penghui;BIAN Sishan;TENG Jiawen(Shandong University of Traditional Chinese Medicine,Jinan 250000,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]山东中医药大学,山东济南250000 [2]山东中医药大学附属医院,山东济南250014

出　　处：《中医药信息》2023年第6期38-43,53,共7页Information on Traditional Chinese Medicine

基　　金：山东省自然科学基金面上项目(2020MH359)。

摘　　要：目的:探讨淫羊藿苷(ICA)调节PTEN激活成骨基因表达缓解大鼠骨质疏松症(OP)的相关作用机制。方法:术中收集患骨质疏松症和骨质正常者股骨骨髓8mL,诱导骨髓间质干细胞(BMSCs)成骨分化14d后,采用qRT-PCR检测PTEN的表达;随后选取7周龄的雌性大鼠36只,采用经典的雌性大鼠卵巢切除法复制绝经后骨质疏松症大鼠模型,术后12周鉴定为骨质疏松模型后开始给药干预,在同等饲养条件下造模大鼠以ICA处理或/和抑制PTEN表达治疗10周后,采取颈椎脱臼法处死大鼠,取双侧股骨干骺端骨组织,采用qRT-PCR法检测大鼠骨组织中PTEN及成骨基因(RUNX2、OPN)的表达、Western blot法检测成骨标志蛋白(RUNX2、OPN)的表达,并制成石蜡切片,采用IHC染色观察PTEN在细胞中的表达、HE染色观察骨组织微观结构。结果:PTEN在OP患者骨组织中高表达,诱导BMSCs成骨分化14d后表达被抑制,造模组大鼠骨组织中PTEN表达也较高,IHC染色见细胞中PTEN表达呈阳性、HE染色骨小梁明显稀疏;ICA处理或/和抑制PTEN表达后PTEN的表达被显著抑制,同时逆转了其对成骨基因表达的抑制作用,HE染色显示骨质疏松大鼠经ICA处理或/和抑制PTEN表达后骨小梁增多、排列紧密,部分新生骨组织形成,可有效缓解大鼠骨质疏松症。结论:ICA可有效抑制PTEN激活成骨基因表达,促进BMSCs成骨分化,缓解大鼠骨质疏松症。Objective:To investigate the action mechanism of icariin(ICA)in alleviating osteoporosis(OP)by regulating PTEN-activated osteogenic genes expression in rats.Methods:Intraoperatively,8 mL femoral bone marrow from patients with osteoporosis or normal bone quality were collected,and the expression of PTEN was detected by qRT-PCR 14 days after induction of osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs);subsequently,36 female rats at 7 weeks of age were selected,and the classic female rat ovariectomy method was used to replicate the postmenopausal osteoporosis rat model.12 weeks after postoperative identification as an osteoporosis model,medication intervention was applied.After 10 weeks of treatment with ICA or/and inhibition of PTEN expression in the same feeding conditions,the rats were executed by cervical dislocation method,and the expression of PTEN and osteogenic genes(RUNX2,OPN)in the bone tissues were detected by qRT-PCR technique.The expressions of osteogenic marker proteins(RUNX2,OPN)were detected by Western blot method.The expression of PTEN in cells was observed by IHC staining after paraffin sections preparation and bone microstructure was observed by HE staining.Results:PTEN was highly expressed in bone tissue of OP patients,and its expression was inhibited 14 days after induction of osteogenic differentiation in BMSCs,and PTEN expression was also higher in bone tissues of the modeled rats.IHC staining showed positive expression of PTEN in cells,and bone trabeculae were sparse in HE staining.The expression of PTEN was significantly inhibited after ICA treatment or/and inhibition of PTEN expression,which simultaneously reversed its inhibition effect on osteogenic genes.HE staining showed that the bone trabeculae in osteoporotic rats were increased and tightly arranged after ICA treatment or/and inhibition of PTEN expression,and some new bone tissues were formed,which effectively alleviated osteoporosis in rats.Conclusion:ICA can effectively inhibit PTEN-activated osteogenic genes

关 键 词：淫羊藿苷 骨质疏松症 PTEN 成骨分化 PTEN/PI3K/Akt信号通路 

分 类 号：R285.5[医药卫生—中药学]