【摘要】 目的 探讨毛蕊花糖苷（VB）通过改善线粒体功能抑制脂多糖（LPS）诱导的骨骼肌炎性应激反应的效应，并探索其保护骨骼肌的机制。方法 (1)C57BL/6J小鼠随机分为正常组、模型组及中药组：正常组腹腔注射0.9%氯化钠溶液，连续干预7 d,1次/d；模型组腹腔注射0.9%氯化钠溶液，连续干预6 d，第7天腹腔注射5 mg/kg LPS溶液，1次/d；中药组腹腔注射150 mg/kg VB溶液，连续干预6 d，第7天腹腔注射5 mg/kg LPS溶液，1次/d。检测小鼠最大抓力和腓肠肌湿质量/体质量比值。(2)C2C12小鼠成肌细胞随机分为正常组、模型组及中药-低、中、高剂量组：正常组常规培养；模型组和各中药组在培养基中加终浓度为200μg/L的LPS；各中药组中加VB，终浓度分别为0.1μmol/L、1μmol/L、10μmol/L。细胞计数试剂盒-8(CCK-8)检测细胞增殖率；酶联免疫吸附分析（ELISA）法检测白细胞介素-6(IL-6);Western blot法检测结蛋白（Desmin）及线粒体融合蛋白2(MFN2)表达；免疫荧光染色法检测Desmin、核因子κB(NF-κB)、MFN2表达。结果 (1)与正常组比较，模型组小鼠最大抓力、腓肠肌湿质量/体质量比值减小（P<0.01）；与模型组比较，VB干预后小鼠最大抓力、腓肠肌湿质量/体质量比值增加（P<0.01,P<0.05）。(2)在C2C12细胞中，与正常组比较，模型组6 h、12 h、24 h细胞增殖率降低（P<0.05,P<0.01）,IL-6水平升高（P<0.01）。24 h后胞内MFN2及线粒体Desmin表达水平降低（P<0.05,P<0.01），核内NF-κB水平升高（P<0.01）；与模型组比较，1μmol/L VB干预6 h, 12 h, 24 h后细胞增殖率升高（P<0.05,P<0.01），干预12 h、24 h后IL-6水平降低（P<0.01），干预24 h后胞内MFN2、线粒体Desmin表达水平上升（P<0.05），核内NF-κB水平减弱（P<0.05）。结论 VB通过改善线粒体功能，下调NF-κB表达，抑制炎症因子过度合成，从而减缓骨骼肌萎缩。更多还原

【Abstract】 Objective To explore whether verbascoside(VB) alleviates lipopolysaccharide(LPS)-induced inflammatory stress in skeletal muscle by enhancing mitochondrial function, and explore its mechanism of protecting skeletal muscles. Methods(1) C57BL/6J mice were randomly divided into normal, model, traditional Chinese medicine(TCM) groups. The normal group was intraperitoneally injected with 0.9% sodium chloride solution for 7 consecutive days, once a day. The model group was intraperitoneally injected with 0.9% sodium chloride solution for 6 consecutive days, and on the 7th day, LPS solution 5 mg/kg was intraperitoneally injected, once a day. In the TCM group, 150 mg/kg VB solution was intraperitoneally injected for 6 consecutive days, and on the 7th day, 5 mg/kg LPS solution was intraperitoneally injected, once a day. Grip strength and the ratio of wet weight to body weight of the gastrocnemius muscle(GW/BW ratio) were measured.(2)C2C12 mouse myoblast cells were divided into normal, model, as well as low-, medium-and high-dose TCM groups. The normal group was cultured routinely; The model and TCM groups were cultured in medium supplemented with LPS(200 μg/L); VB was added to the TCM groups at final concentrations of 0.1 μmol/L, 1 μmol/L and 10 μmol/L, respectively. Cell Counting Kit-8(CCK-8) was used to detect cell proliferation; Enzyme-linked immunosorbent assay(ELISA) was used to measure IL-6 levels; Western blot was used to analyze Desmin and mitochondrial fusion protein 2(MFN2) expressions; Immunofluorescence staining was used to detect Desmin, NF-κB, MFN2 expressions. Results(1)Compared with the normal group, mice in the model group had significantly lower grip strength and GW/BW ratio(P<0.01); After VB intervention, the maximum grip strength and GW/BW ratio of mice increased compared with the model group(P<0.01, P<0.05).(2)In C2C12 cells, compared with the normal group, the model group showed decreased proliferation at 6 h, 12 h, 24 h(P<0.05, P<0.01), increased IL-6 level(P<0.01). After 24 h, mitochondrial MFN2 and intracellular Desmin expression levels decreased(P<0.05), intranuclear NF-κB level increased(P<0.01). Compared with the model group, 1 μmol/L VB intervention increased cell proliferation at 6 h, 12 h, 24 h(P<0.05, P<0.01, P<0.01), decreased IL-6 level at 12 h, 24 h(P<0.01); After intervention for 24 h, mitochondrial MFN2 and intracellular Desmin expression levels increased(P<0.05), intranuclear NF-κB level decreased(P<0.01). Conclusion VB improves mitochondrial function, reduces NF-κB expression, inhibits excessive synthesis of inflammatory factors, thereby slowing down skeletal muscle atrophy.更多还原