基于BMP/Runx2/Osx信号通路研究淫羊藿总黄酮改善绝经后骨质疏松模型大鼠的作用机制
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篇名: 基于BMP/Runx2/Osx信号通路研究淫羊藿总黄酮改善绝经后骨质疏松模型大鼠的作用机制
TITLE: Study on the Effect and Mechanism of Total Flavonoids of Epimedium brevicornu on Postmenopausal Osteoporosis Model Rats Based on BMP/Runx 2/Osx Signaling Pathway
摘要: 目的:基于骨形态发生蛋白(BMP)/Runt相关转录因子2(Runx2)/成骨细胞特异性转录因子(Osx)信号通路研究淫羊藿总黄酮对绝经后骨质疏松(PMOP)模型大鼠的影响,从而明确其防治骨质疏松症(OP)的作用机制。方法:将50只大鼠按体质量分层法随机分为假手术组,模型组,淫羊藿总黄酮低、高剂量组[265、530mg/(kg·d)]和雌二醇组[0.09mg/(kg·d)],每组10只。除假手术组大鼠行假手术外,其余各组大鼠均采用卵巢摘除去势法建立PMOP模型。大鼠造模成功并进行正常饲养2个月后开始灌胃给药,每天给药1次,连续给药84d;假手术组和模型组大鼠灌胃等量生理盐水。末次给药结束后,测定各组大鼠右侧下肢股骨和椎骨的骨密度(BMD)以及股骨骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.Sp);采用酶联免疫吸附法测定大鼠血清中钙离子、骨钙素、Ⅰ型原胶原N端前肽(P1NP)的水平;采用苏木精-伊红染色法观察股骨的病理学变化;分别采用实时荧光定量-聚合酶链式反应法和Westernblotting法检测大鼠骨组织中BMP、Runx2、Osx的mRNA及蛋白的表达水平。结果:与假手术组比较,模型组大鼠股骨、椎骨BMD,血清中钙离子、骨钙素、P1NP水平,股骨Tb.N和Tb.Th以及股骨组织中BMP、Runx2、OsxmRNA及蛋白表达水平均显著降低(P<0.01),股骨Tb.Sp显著升高(P<0.01);骨小梁组织结构紊乱,断裂明显。与模型组比较,各给药组大鼠上述指标均显著改善(P<0.05或P<0.01),并且淫羊藿总黄酮的作用具有剂量依赖性(P<0.05);骨小梁数目增多,排列整齐,结构较为完整。结论:淫羊藿总黄酮改善OP的作用机制可能与促进BMP/Runx2/Osx信号通路活性有关。
ABSTRACT: OBJECTIVE:To study the effect of total flavonoids of Epimedium brevicornu on postmenopausal osteoporosis (PMOP)model rats based on BMP/Runx 2/Osx signaling pathway so as to confirm the mechanism of preventing and treating osteoporosis(OP). METHODS :By body mass stratification ,50 rats were randomly divided into sham operation group ,model group,E. brevicornu total flavonoids low-dose and high-dose groups [265,530 mg/(kg·d)],estradiol group [0.09 mg/(kg·d)], with 10 rats in each group. Except that sham operation group underwent sham operation ,PMOP model was established by ovariectomy and castration in other groups. After modeling ,they were given normal diet for 2 months and then given relevant medicine intragastrically for consecutive 84 d,once a day ;rats in sham operation group and model group were given equal volume of normal saline. After last medication ,bone mineral density (BMD)of femur and vertebrae of the right lower limb ,the number of trabecular bone (Tb.N),trabecular bone thickness (Tb.Th)and trabecular separation (Tb.Sp)of femur were determined in each group. The serum levels of Ca 2+,OC and P 1NP in serum were detected by ELISA. HE staining was used to observe pathological changes of femur. mRNA and protein expressions of BMP ,Runx2 and OSX in bone tissue were detected by RT-PCR and Western blotting. RESULTS :Compared with sham operation group ,BMD of femur and vertebrae ,serum levels of Ca 2 +,OC and P 1NP, Tb.N and Tb.Th of femur ,mRNA and protein expression of BMP ,Runx2 and Osx in femur were decreased significantly in model group,while Tb.Sp of femur was increased significantly (P<0.01);the structure of trabecular bone was disordered and the fracture was obvious. Compared with model group ,above indexes of rats in administration groups were improved significantly (P< 0.05 or P<0.01),and the effect of E. brevicornu total flavonoids was dose-dependent (P<0.05);the number of trabecular bone increased, arranged orderly and the structure was more brevicornu total flavonoids can improve OP ,the mechanism of which may be associated with com promoting the activity of BMP/Runx 2/Osx signaling pathway.
期刊: 2020年第31卷第19期
作者: 徐众华,莫雨晴,周驰
AUTHORS: XU Zhonghua ,MO Yuqing ,ZHOU Chi
关键字: 淫羊藿总黄酮;骨质疏松症;骨形态发生蛋白;Runt相关转录因子2;成骨细胞特异性转录因子;大鼠
KEYWORDS: Epimedium brevicornu total flavonoids ;
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