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1.上海中医药大学中西医结合学院(上海 201203)
2.上海交通大学医学院附属上海儿童医学中心中医科(上海 200127)
3.上海中医药大学中药学院(上海 201203)
4.中国科学院上海药物研究所药物发现与设计中心(上海 201203)
汤泽玲,女,硕士研究生,主要从事中医药延缓衰老与防治老年病的研究工作
张勇,副研究员;E-mail:zhangyong109@simm.ac.cn
龚张斌,教授,博士研究生导师;E-mail:zhangbingong @126.com
收稿日期:2024-12-31,
纸质出版日期:2025-09-10
移动端阅览
汤泽玲,陈莉萍,马梦玲,等.毛蕊花糖苷改善线粒体功能抑制骨骼肌炎性应激反应[J].上海中医药杂志,2025,59(9):68-74.
TANG Zeling,CHEN Liping,MA Mengling,et al.Verbascoside inhibits inflammatory stress response in skeletal muscles through promoting mitochondrial function[J].Shanghai Journal of Traditional Chinese Medicine,2025,59(9):68-74.
汤泽玲,陈莉萍,马梦玲,等.毛蕊花糖苷改善线粒体功能抑制骨骼肌炎性应激反应[J].上海中医药杂志,2025,59(9):68-74. DOI: 10.16305/j.1007-1334.2025.z20241231001.
TANG Zeling,CHEN Liping,MA Mengling,et al.Verbascoside inhibits inflammatory stress response in skeletal muscles through promoting mitochondrial function[J].Shanghai Journal of Traditional Chinese Medicine,2025,59(9):68-74. DOI: 10.16305/j.1007-1334.2025.z20241231001.
目的
2
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
2
①C57BL/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。②C2C12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200 μg/L的LPS;各中药组中加VB,终浓度分别为0.1 μmol/L、1 μmol/L、10 μmol/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
2
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,1 μmol/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
2
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。
Objective
2
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
2
①C57BL/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. ②C2C12 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
2
①Compared 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). ②In 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
2
VB improves mitochondrial function, reduces NF-κB expression, inhibits excessive synthesis of inflammatory factors, thereby slowing down skeletal muscle atrophy.
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