补肾生骨汤对股骨头无菌性坏死大鼠的影响  

Efficacy of Bushen Shenggu Decoction on miR-135b-5p/RUNX2 Signaling Axis in Rats with Aseptic Necrosis of the Femoral Head

作　　者：艾力江·阿斯拉 梁治权[2] 李雷疆[2] 邓迎杰[2] 廖军[2] 王继先[2] Ailijiang Asila;LIANG Zhiquan;LI Leijiang;DENG Yingjie;LIAO Jun;WANG Jixian(Xinjiang Uygur Autonomous Region Institute of Traditional Chinese Medicine,Urumqi 830000,China;Department of Orthopedics,Affiliated Hospital of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830000,China)

机构地区：[1]新疆维吾尔自治区中医药研究院,乌鲁木齐830000 [2]新疆医科大学附属中医医院骨科

出　　处：《中国中医骨伤科杂志》2023年第1期7-12,共6页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：新疆维吾尔自治区自然科学基金资助项目(2019D01C178)。

摘　　要：目的:探讨补肾生骨汤调节miR-135b-5p/Runt相关基因2(RUNX2)信号轴对股骨头无菌性坏死(ONFH)大鼠的影响。方法:将SD大鼠随机分为假手术(Sham)组、ONFH组、低/中/高剂量补肾生骨汤(0.13,0.26,0.52 g/mL,L/M/H-补肾生骨汤)组、H-补肾生骨汤+miR-135b-5p agomir组、H-补肾生骨汤+agomir-NC组,采用液氮冷冻法制备ONFH大鼠模型;苏木精-伊红(HE)染色观察股骨头组织病理改变;检测血浆中纤维蛋白原(FIB)、血管内皮生长因子(VEGF)、一氧化氮合成酶(NOS)、骨钙素(BGP)水平;RT-qPCR仪、Western Blot法分别检测股骨头组织中miR-135b-5p、RUNX2表达水平;双荧光素酶实验分析miR-135b-5p与RUNX2的靶向关系。结果:与Sham组比较,ONFH组股骨头组织发生区域性缺损,骨小梁稀疏、行走不规则,空骨陷窝增多明显;血浆中FIB表达升高,VEGF、NOS、BGP表达降低(P<0.05);经L/M/H-补肾生骨汤干预后,上述情况均改善,且经H-补肾生骨汤干预改善更明显;在H-补肾生骨汤干预的基础上增加miR-135b-5p agomir干预后,H-补肾生骨汤干预的改善作用被削弱。与Sham组比较,ONFH组股骨头组织中miR-135b-5p表达升高,RUNX2表达降低(P<0.05);经H-补肾生骨汤干预后,miR-135b-5p表达降低,RUNX2表达升高(P<0.05);在H-补肾生骨汤干预的基础上增加miR-135b-5p agomir干预后,miR-135b-5p表达升高,RUNX2表达降低(P<0.05)。双荧光素酶实验结果显示,miR-135b-5p与RUNX2存在靶向关系。结论:补肾生骨汤能促进ONFH大鼠股骨头组织修复,可能与下调miR-135b-5p进而靶向上调RUNX2有关。Objective:To investigate the efficacy of Bushen Shenggu decoction on microRNA(miR)-135b-5p/Runt-related transcription factor 2(RUNX2) signaling axis in rats with aseptic necrosis of the femoral head(ONFH).Methods:SD rats were randomly divided into Sham operation group, ONFH group, low/medium/high-dose Bushen Shenggu decoction(0.13,0.26,0.52 g/mL) groups, H-Bushen Shenggu decoction + miR-135b-5p agomir group, and H-Bushen Shenggu decoction + agomir-NC group.The ONFH rat model was established by liquid nitrogen freezing method.Histopathological changes of femoral head were observed by hematoxylin-eosin(HE) staining.The levels of fibrinogen(FIB),vascular endothelial growth factor(VEGF),nitric oxide synthase(NOS) and osteocalcin(BGP) in plasma were detected.RT-qPCR and Western Blot were performed to detect the expression levels of miR-135b-5p and RUNX2 in femoral head tissue, respectively.The targeting relationship between miR-135b-5p and RUNX2 was analyzed by dual luciferase experiments.Results:Compared with the Sham group, the femoral head tissue in the ONFH group had regional defects, sparse and irregular trabecular bone, and greatly more empty bone lacunae;the expression of FIB in plasma increased, while the expression of VEGF,NOS and BGP decreased(P<0.05).After the intervention of L/M/H-Bushen Shenggu decoction, the above conditions were improved, and the improvement was more obvious after the intervention of H-Bushen Shenggu decoction.After adding miR-135b-5p agomir on the basis of H-Bushen Shenggu decoction intervention, the improvement efficacy of H-Bushen Shenggu decoction intervention was weakened.Compared with the Sham group, the expression of miR-135b-5p in the femoral head tissue of the ONFH group increased, and the expression of RUNX2 decreased(P<0.05).After the intervention of H-Bushen Shenggu decoction, the expression of miR-135b-5p decreased and the expression of RUNX2 increased(P<0.05),the expression of miR-135b-5p increased and the expression of RUNX2 decreased after agomir intervention on the basi

关 键 词：补肾生骨汤 股骨头无菌性坏死 骨钙素 血管内皮生长因子 一氧化氮合成酶 

分 类 号：R-33[医药卫生]