陆军军医大学学报2026，Vol.48Issue(1)：42-53,12.DOI:10.16016/j.2097-0927.202511078
高寒环境通过诱导骨骼肌萎缩与纤维化协同加重骨关节炎病理进程
Cold exposure exacerbates osteoarthritis via synergistic muscle atrophy and fibrosis
摘要
Abstract
Objective To investigate the impact of a high-altitude and cold environment on skeletal muscle function and the pathological processes of osteoarthritis(OA),to elucidate the interaction mechanisms between skeletal muscle atrophy,fibrosis,and joint degeneration under cold condition,aiming to found a theoretical basis for the health protection of the musculoskeletal system in military personnel stationed in high-altitude cold regions.Methods Destabilization of the medial meniscus(DMM)was established on C57BL/6 mice(11 weeks old)by cutting the attaching point at the anterior intercondylar region where anterior cruciate ligament blends with the medial meniscus.Forty male C57BL/6 mice(11 weeks old)were stratified by 2 postoperative time points(8 and 12 weeks).At each time point,the mice were divided randomly divided into 4 groups(n=5):22℃sham operation(22℃Sham),4℃sham operation(4℃Sham),22℃DMM,and 4℃DMM.From 2 weeks after DMM modelling,the 4℃Sham and 4℃DMM groups were placed in a cold chamber[4℃,humidity(50±5)%]for 12 h per day until the respective endpoint,meanwhile,the 22℃Sham and 22℃DMM groups were kept at room temperature.At 8 and 12 weeks after intervention,the following assessments were carried out:① Behavioral tests:Gait parameters were quantified using the CatWalk system;Limb load-bearing capacity was evaluated with bipedal balance test;Spontaneous activity distance and time were recorded in the open field test;Twitch force and tetanic force in the quadriceps were measured,and the force-frequency curve was plotted.② Histopathological analysis:Quadriceps and knee joint tissues were harvested,embedded in paraffin,sectioned and stained with HE and safranin O-fast green to assess synovial inflammatory cell infiltration,cartilage surface integrity,and OARSI score from Osteoarthritis Research Society International System.Additionally,Masson's trichrome staining was applied to observe collagen deposition in muscle fibers.③ Molecular detection:Immunofluorescence assay for dystrophin antibody was used to label muscle fiber boundaries and calculate cross-sectional area;Immunohistochemistry assay was employed to determine the expression levels of IL-1β and TNF-α in synovial tissue.Results ① Progressive deterioration of behavioral function:Compared with the 22℃DMM group,the 4℃DMM group exhibited significantly exacerbated functional impairments at both 8 and 12 weeks after intervention,in a time-dependent worsening trend.a.Loss of gait coordination:At 8 weeks,the relative stance phase time of the right hindlimb was shorten by 0.113(P<0.001),print area was reduced by 0.102(P<0.05),swing phase time was extended by 0.115(P<0.05),and single-limb stand time was prolonged by 0.146(P<0.01).These abnormalities still existed at 12 weeks,with the difference of print area expanding to 0.154(P<0.05)and that of swing phase time maintaining an elevated magnitude of 0.113(P<0.05).b.Decline in weight-bearing capacity:Percentage of right paw weight-bearing was decreased by 2.777%at 8 weeks(P<0.001),with the deficit further expanding to 5.156%at 12 weeks(P<0.000 1).c.Suppression of spontaneous activity:Total locomotor distance was decreased by 1 069 cm at 8 weeks(P<0.000 1)and remained reduced by 429.3 cm at 12 weeks(P<0.05).② Exacerbation of joint histopathological damage:Cold exposure combined with DMM surgery significantly aggravated synovial inflammation and cartilage destruction.a.Synovial pathology:The 4℃DMM group displayed increased synovial pathological score of 2.133 at 8 weeks(P<0.01)and then 2.833 at 12 weeks(P<0.001)relative to the 22℃DMM group,indicating a sustained inflammatory response.b.Cartilage degeneration:The 4℃DMM group showed the maximum score of OARSI elevated by 1.000(P<0.05)and total score by 2.000(P<0.01)at 8 weeks,while,at 12 weeks,the maximum score remained elevated by 0.750,and the total score maintained a differential trend of 2.000.c.Upregulation of pro-inflammatory factors:The synovial expression area of IL-1β in the 4℃DMM group was increased by 1.047%(P<0.05)and 1.892%(P<0.05)at 8 and 12 weeks,respectively,and that of TNF-α expression was increased by 3.850%(P<0.000 1)and 3.806%(P<0.000 1),separately at the same time points.③ Composite impairment of skeletal muscle structure and contractile function:Cold intervention induced triple pathological changes in quadriceps encompassing atrophy,mechanical performance decline,and fibrotic remodeling.a.Depletion of muscular force reserve:Twitch force was decreased by 34.77 mN·m/kg(P<0.01)and 52.87 mN·m/kg(P<0.05)at 8 and 12 weeks,respectively;Tetanus force was reduced by 115.0 mN·m/kg(P<0.05)and 138.3 mN·m/kg(P<0.000 1)at the same time points,with a downward shift of the force-frequency curve.b.Muscle atrophy:The mass ratio of quadriceps was decreased by 0.076%(P<0.05)and 0.102%(P<0.000 1),and the muscle fiber cross-sectional area(CSA)was reduced by 261.3 μm²(P<0.05)and 233.5 μm²(P<0.01)at 8 and 12 weeks,respectively,accompanied with an increased proportion of small-diameter fibers.c.Fibrotic deposition:Relative collagen fiber area in muscle tissue was increased by 4.072%(P<0.01)and 2.238%(P<0.05)at 8 and 12 weeks,respectively,suggesting abnormal extracellular matrix remodeling.Conclusion High-altitude cold environment not only directly aggravates synovial inflammation and cartilage degeneration in OA but also further compromises joint structure and function by inducing quadriceps atrophy,reduced muscle strength,and fibrosis.关键词
军事医学/高寒环境/骨关节炎/骨骼肌/肌肉萎缩/滑膜炎症Key words
military medicine/cold environment/osteoarthritis/skeletal muscle/muscle atrophy/synovial inflammation分类
医药卫生引用本文复制引用
藤培煜,谢杨丽,陈林,李松,龙尘宇,黄俊兰,杨鹏,苏楠,罗小庆,杨京,张若彬..高寒环境通过诱导骨骼肌萎缩与纤维化协同加重骨关节炎病理进程[J].陆军军医大学学报,2026,48(1):42-53,12.基金项目
国家自然科学基金重大项目(82394444) （82394444）
重庆市科卫联合项目(2024DBXM008) （2024DBXM008）
后勤科研重点项目(BLJ22J014) Supported by the Major Program of National Natural Science Foundation of China(82394444),the Joint Project of Chongqing Science and Technology Commission and Chongqing Health Commission(2024DBXM008),and the Key Logistics Research Project(BLJ22J014). （BLJ22J014）
