摘要
目的探究骨细胞连接蛋白43(connexin43,Cx43)通道在力学过载对小鼠膝关节影响中的作用。方法对15周龄骨细胞特异Cx43转基因小鼠(R76W:间隙连接功能抑制而半通道功能促进;Δ130⁃136:间隙连接和半通道都被抑制)及同窝野生型小鼠左膝关节施加单次应变(ε=1.76462),右膝关节作为对照。1周后分析小鼠步态,评价小鼠膝关节骨与软骨情况。抑制MLO⁃Y4骨细胞Cx43通道功能后进行循环拉伸应变,检测相关因子表达。收集条件培养基培养ATDC5软骨细胞,检测相关基因表达和成骨分化能力。结果步态分析显示,Cx43通道抑制没有明显影响小鼠步态。但转基因小鼠关节股骨端OARSI评分显著增加,骨小梁面积占比更小。抑制MLO⁃Y4细胞Cx43通道降低了ADAMTS5和II型胶原表达,增强其成骨分化的能力。结论骨细胞Cx43通道能够响应力学过载,影响软骨细胞关键基因表达,并参与其成骨分化过程的调控,影响软骨对力学过载的响应。研究结果为骨关节炎的防治提供新思路。
Objective To investigate the role of osteocytic connexin43(Cx43)⁃formed channels after being submitted to mechanical overloading on knee joint.Methods The strain was applied to left knee joint of 15⁃week Cx43 transgenic mice(R76W:gap junctions were blocked and hemichannels were promoted;Δ130⁃136:both hemichannels and gap junctions were blocked)and their wild⁃type littermates,and right knee joint was as control.A week later,the mice gait was analyzed to evaluate the structure of subchondral bone and articular cartilage.After inhibition of Cx43 channels,MLO⁃Y4 cells were submitted to 15%cyclic tensile strain,and the expression of related factors were detected.The supernatant was collected to culture ATDC5 cells,and the gene expression and the osteogenic differentiation ability were detected in ATDC5 cells.Results Gait analysis showed that inhibition of Cx43 channels did not significantly affect the mice gait.However,the femoral OARSI score increased significantly and the ratio of trabecular bone area decreased in transgenic mice.Conditional medium from MLO⁃Y4 cells with inhibition of Cx43 channels reduced the expression of ADAMTS5 and COL2 in ATDC5 cells,and enhanced the osteogenic differentiation ability.Conclusions The osteocytic Cx43 channels can respond to mechanical overloading,affect the expression of key genes and is involved in osteogenic differentiation process in chondrocytes,thus affects cartilage structure in response to mechanical overloading.The study will provide new ideas for the prevention and treatment of osteoarthritis.
作者
陈家辉
杨保强
宁凯婷
续惠云
CHEN Jiahui;YANG Baoqiang;NING Kaiting;XU Huiyun(School of Life Sciences,Northwestern Polytechnical University,Xi’an 710072,China)
出处
《医用生物力学》
CAS
CSCD
北大核心
2022年第5期881-887,966,共8页
Journal of Medical Biomechanics
基金
国家自然科学基金项目(81772409)。
