补肾活血中药龙鳖制剂干预野生型TDP43介导的骨性关节炎软骨细胞病变的作用研究  

Wild Type TDP-43 Mediated Chondrocyte Pathology by Intervention of TCM Longbie Preparation in Osteoarthritis

作　　者：黄永明[1] 黄启明 曹振武 苏海涛[1] 麦秀钧[1] 冯恩辉 

机构地区：[1]广州中医药大学第二临床医学院,广东广州510405 [2]怡诺博(北京)生物医学技术有限公司,北京100088

出　　处：《辽宁中医杂志》2018年第1期96-100,I0003,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81273781,81473698)

摘　　要：目的：分析补肾活血中药龙鳖对TDP-43引起骨性关节炎应急反应的软骨细胞病变的干预作用。方法：通过构建TDP43慢病毒载体并转染人软骨细胞,流式细胞检测分析凋亡水平,连续细胞计数分析其细胞增殖水平,荧光定量PCR检测关键基因表达水平;采用软骨细胞培养液中加入低、中、高剂量龙鳖进行干预,分析TDP43慢病毒转染软骨细胞后的凋亡和增殖生长水平。结果：人软骨细胞转染包装好的TDP43慢病毒后,高表达TDP43基因和I型胶原蛋白α1。TDP43慢病毒转染的软骨细胞体外连续培养软骨细胞7 d后,软骨细胞出现晚期凋亡和死亡现象而抑制了细胞增殖。龙鳖加入培养后软骨细胞呈生长趋势,TDP43慢病毒转染的软骨细胞凋亡显著下降而显著提高了其增殖水平。结论：转染TDP43慢病毒软骨细胞表达TDP43引起了细胞凋亡和抑制其增殖,TDP43可能在骨性关节炎发病过程中取到负调控的作用,而中药龙鳖制剂可显著改善这一过程。Objective： To study the intervening role of traditional Chinese medicine（TCM） Longbie Preparation on TAR DNA-binding protein-43（TDP43） involved in osteoarthritis（OA） progression. Methods： We constructed TDP43 lentivirus vector which transfected human chondrocytes（HCs） and investigated that the expressive level of target genes in the HCs by qRT-PCR. The cell function assay was used to evaluate the apoptosis of human chondrocytes（HCs） by Annexin V and PI staining,and its proliferation level by cells counting. By adding low,middle and high dose of Longbie Preparation in HCs culture,TDP43 lentivirus transfected HCs were identified proliferative and apoptotic levels. Results： The HCs highly expressed TDP43 and COL I α1 gene after transfecting with TDP43 lentivirus,and maintained its characterizations. The TDP43 overexpression promoted HCs later apoptosis and death but inhibited HCs proliferation after culturing at 7 days. The HCs emerged growth tendency after adding Longbie preparation in HCs culturing medium. By compared with no Longbie Preparation,the apoptotic level of TDP43-HCs was significantly reduced in high-dose Longbie Preparation and increased proliferative level. Conclusion： The HCs induced apoptosis and inhibited proliferation after transfecting with TDP43 genes. These data demonstrate for the first time that TDP43 negatively mediated the pathogenesis of OA. The TCM Longbie Preparation contributed to promoting the OA onset,a potential approach for the treatment of OA.

关 键 词：龙鳖制剂(胶囊) 骨性关节炎 软骨细胞 TARDNA结合蛋白43 补肾活血中药 

分 类 号：R285.5[医药卫生—中药学]