| 秋超,唐毅华,陈亿忞,陈诗佳,张丽萍.炙马钱子对重症肌无力小鼠血清MMP-3 及抗MuSK抗体影响[J].浙江中西医结合杂志,2023,33(4): |
| 炙马钱子对重症肌无力小鼠血清MMP-3 及抗MuSK抗体影响 |
| Effect of Nux Vomica on Serum Anti-Musk Antibody in Myasthenia Gravis Mice |
| 投稿时间：2022-08-27 修订日期：2023-03-01 |
| DOI： |
| 中文关键词: 小鼠 重症肌无力 MuSK抗体 AG490 MMP-3 |
| 英文关键词:Mouse Myasthenia Gravis Musk antibody AG490 MMP-3 |
| 基金项目:浙江省自然科学基金(No.LQ20H270015);浙江省医药卫生科技计划(2019RC058) |
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| 中文摘要: |
| 目的 探讨炙马钱子通过调节基质金属蛋白酶3（MMP-3）致使重症肌无力发病机制。方法 45只8周雄性C57BL/6J小鼠，麻醉后，随机抽取37只模型组小鼠，共用40 μg的肌肉特异性酪氨酸激酶（MuSK）乳化在100 μL磷酸盐缓冲溶液（PBS）和100 μL完全弗氏佐剂（CFA）腹腔、后脚注射，剩余8只正常组注射100 μL PBS和100 μL CFA佐剂，并24 h内注射环磷酰胺（300 mg/kg，溶解在0.9%的生理盐水中配置成10 mg/mL的终浓度）抑制免疫抵抗。30 d补充注射1次，通过小鼠体征及行为学等观察确定造模周期。在造模成功当天，将小鼠随机分为正常组、模型组、炙马钱子组、AG490组，每组8只，炙马钱子组予炙马钱子250 mg/（kg·d）连续灌胃30 d；AG490组予炙马钱子250 mg/（kg·d）连续灌胃30 d，AG490 5 mg/（kg·d）连续腹腔注射30 d；正常组和模型组灌胃等量0.9%的生理盐水。采用酶联免疫吸附（ELISA）法检测血清MuSK滴度，蛋白质印迹法（Western blot）检测各组神经肌肉接头处中MMP-3的蛋白表达水平。结果 MuSK滴度水平：正常组MuSK滴度水平最低（3.23±1.89）；与模型组比较，炙马钱子组、AG490组血清MuSK滴度显著降低[（242.12±24.69）、（133.68±27.27）比（856.93±32.44），P<0.05]，且随着给药时间的延长，炙马钱子组、AG490组MuSK滴度水平逐渐下降。MMP-3的蛋白表达水平：正常组MMP-3的蛋白表达水平最低（1.00±0.07）；与模型组比较，炙马钱子组、AG490组MMP-3的蛋白表达水平逐渐下降[（1.60±0.10）、（1.27±0.13）比（2.30±0.11），P<0.05]。结论 炙马钱子可能通过使MMP-3合成减少，降低MuSK抗体，达到治疗重症肌无力的作用。 |
| 英文摘要: |
| Objective To explore the pathogenesis of myasthenia gravis caused by the regulation of matrix metalloproteinase 3 (MMP-3) by Sunburn Strychnine. Methods A total of 45 8-week male C57BL/6J mice were anesthetized and 37 mice in the model group were randomly selected. 40 μg muscular-specific tyrosine kinase (MuSK) was emulsified and injected into the abdominal cavity and hind foot with 100 μl phosphate buffer solution (PBS) and 100 μl complete Freundsen's adjuvant (CFA). The remaining 8 normal groups were injected with 100 μl PBS and 100 μl CFA adjuvant, and cyclophosphamide (300 mg/kg dissolved in 0.9% saline to a final concentration of 10 mg/ml) was injected within 24 hours to suppress immune resistance. The mice were injected once every 30 days, and the model period was determined by observing the signs and behavior of mice. On the day of successful modeling, mice were randomly divided into normal group, model group, broiled strychnos group and AG490 group, with 8 mice in each group. The broiled strychnos group was given broiled strychnos (250 mg/kg/d) by gavage for 30 days. Group AG490 was given Strychnine moxibusi (250 mg/kg/d) by gavage for 30 days, and group AG490 (5 mg/kg/d) by intraperitoneal injection for 30 days. The normal group and the model group were gavaged with the same volume of 0.9% normal saline. The serum MuSK titer was detected by enzyme-linked immunosorbent assay (ELISA), and the protein expression level of MMP-3 in the neuromuscular junction was detected by Western-blot. Results MuSK titer level: The MuSK titer level in normal group was the lowest (3.23±1.89). Compared with the model group, the serum MuSK titer of brochuria strychnine group and AG490 group was significantly decreased [(242.12±24.69), (133.68±27.27) vs (856.93±32.44), P< 0.05], and with the prolongation of administration time, the levels of MuSK titers in Strychnos moxibusi and AG490 groups gradually decreased. MMP-3 protein expression level：The protein expression level of MMP-3 in normal group was the lowest (1.00±0.07). Compared with the model group, the protein expression level of MMP-3 in broilers and AG490 groups decreased gradually [(1.60±0.10), (1.27±0.13) vs (2.30±0.11), P<0.05]. Conclusion Sunburn Strychnine may reduce the synthesis of MMP-3 and reduce MuSK antibody to achieve the therapeutic effect of myasthenia gravis. |
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