项目名称： 梅花鹿鹿茸多肽调控骨关节炎软骨细胞TGF-β信号通路的物质基础和分子机制研究

项目编号： No.81503177

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 张郑瑶

作者单位： 大连理工大学

项目金额： 18万元

中文摘要： 骨性关节炎是一种以关节软骨代偿失调为病理特征的慢性关节病，常见于老年人且发病率逐年上升。鹿茸是哺乳动物中唯一可再生的骨类组织，从梅花鹿鹿茸中提取的多肽明显加速骨折大鼠骨痂形成，缩短愈合时间，促进软骨细胞增殖，但精确作用机制尚不清晰。本研究发现，鹿茸多肽主要成分nVAP单体对骨性关节炎软骨细胞的损伤有修复与保护作用，可能与影响TGF-β信号通路转导有关。为探讨nVAP单体调控机制，一方面采用酵母双杂交等方法，明确TGF-β及其受体胞外区与nVAP单体作用靶点，观察通路下游分子Smad2/3表达及细胞基质代谢情况，从而阐释nVAP促进软骨细胞增殖和抑制骨赘形成的双重调控效应，为鹿茸的老药新用提供理论支持。另一方面测定nVAP单体作用下MSC和软骨细胞TGF-β相关基因差异表达，筛选参与调控的microRNA，为骨性关节炎的基因治疗提供实验依据。本研究亦可为鹿茸的中药现代化应用提供新思路。

中文关键词： 鹿茸多肽；骨关节炎；软骨细胞；TGF-β信号通路；分子机制

英文摘要： Osteoarthritis is a chronic joint disease of pathological decompensation in articular cartilage, the increasing incidence of which is quite common in elderly people. Velvet antler is a renewable bone tissue, which is totally unique in mammalian. The total polypeptides extracted from fresh Cervus nippon antler could significantly accelerated fracture callus formation in rats, shorten the healing time, ameliorated the biomechanical properties of injured parts, and promoted the proliferation of chondrocytes. However, the exact molecular regulatory mechanism remains unclear. Our previous study demonstrated that total polypeptides had the repairable and protectable capability to rescue osteoarthritis chondrocytes injury. Further research revealed that the isolated nVAP monomer had effected on TGF-β signaling pathway . To identify the effect targets we present experiments including the yeast two-hybrid to elucidate the binding sites between nVAP and TGF-β1/TGF-β2; also nVAP and TβRI-ED/TβRII-ED. Observe the expression of downstream molecule Smad2/3 and the cell matrix degradation. Interpretate the dual regulation effects of nVAP in promoting the proliferation of cartilage cells and the inhibition the formation in osteophyte. Meanwhile, to provide new targets for gene therapy of osteoarthritis, we would detect the differential expression of TGF-β gene after effected by nVAP, and screen the changes of the microRNA who involved in the regulation. Preliminary exploration on the nVAP’s effect mechnism, could establish the theoretical and experimental basis of the modernization of traditional Chinese medicine resources.

英文关键词： Velvet antler polypeptides;osteoarthritis;chondrocyte;TGF-β signaling pathway;molecular mechanism