葛根及其配伍对重症肌无力大鼠骨骼肌线粒体结构的影响(1)
〔摘要〕 目的 探讨葛根及其配伍对重症肌无力大鼠骨骼肌线粒体结构的作用机制。方法 (1)造模：给模型大鼠以腹部、足垫、背部皮下注射200 μL(含R 97-116∶100 μg)免疫乳剂进行造模，30 d和45 d后，分别以同样方法对造模大鼠强化接种，以大鼠Lennon肌力评分≥1级为模型鼠。(2)分组与处理：除空白组外，造模成功鼠随机分为模型组、强的松组、葛根组、葛芪组、葛参组、葛妙组、葛芪参妙组，每组20只。造模成功2周后，给予空白组、模型组蒸馏水灌胃，其他组予相应药物灌胃，连续4周。(3)标本检测：透射电镜观察骨骼肌线粒体超微结构;免疫组化法检测FGF、NGF、IGFs表达。结果 与模型组比较，各组的线粒体形态均有恢复，其中葛芪组和葛芪参妙组治疗后整体形态恢复较好;与模型组比较，各治疗组FGF、NGF、IGFs的阳性表达率都有所升高，差异有统计学意义(P<0.01)。结论 葛根及其配伍可能通过改善肌肉生成调控因子的阳性表达促进线粒体结构的恢复，达到治疗重症肌无力的目的。
〔关键词〕 重症肌无力;葛根配伍;骨骼肌线粒体超微结构;肌肉生成调控因子
〔Abstract〕 Objective To explore the action mechanism of Radix Puerariae and its compatibility on the mitochondrial structure of skeletal muscle in myasthenia gravis rats. Methods (1)modeling： The model rats were established by injecting 200 μL (including r 97-116∶100 μg) immune emulsion into abdomen， foot pad and back subcutaneously. After 30 days and 45 days， the rats were inoculated with the same method， and Lennon score ≥ 1 was used as the model rats. (2)Grouping and treatment： In addition to the blank group， the successfully modeling rats were randomly divided into a model group， a prednisone group， a Radix Puerariae group， a Geqi group， a Geshen group， a Gemiao group and a Geqishenmiao group， with 20 rats in each group. After 2 weeks of successful modeling， the blank group and model group were given distilled water by gavage， and the other groups were given drugs by gavage for 4 weeks. (3)Sample detection： The mitochondrial ultrastructure of skeletal muscle was observed by transmission electron microscope. The expression of FGF， NGF and IGFs was detected by immunohistochemistry. Results Compared with the model group， the mitochondrial morphology of each group recovered， and the whole morphology of the Geqi group and the Geqishenmiao group recovered better after treatment; Compared with the model group， the positive expression rates of FGF， NGF and IGFs in each treatment group increased， and the difference was statistically significant (P<0.01). Conclusion Radix Puerariae and its compatibility may promote the recovery of mitochondrial structure by improving the positive expression of myogenic regulatory factors， so as to achieve the purpose of treating myasthenia gravis.
〔Keywords〕 myasthenia gravis; compatibility of Radix Puerariae; ultrastructure of skeletal muscle mitochondria; regulation factors of muscle formation
重癥肌无力(myasthenia gravis，MG)是一种难治性自身免疫性疾病，以眼肌或全身肌无力为主要表现[1]，到目前为止，其病理机制尚不十分清楚，病因以乙酰胆碱受体(acetylcholine receptor， AchR)受损为主，即神经肌肉接头处突触后膜中因AchR分子数量减少无法产生足够的终板电位而导致肌肉传递功能障碍[2]，最终产生肌无力的症状。中医学认为MG病机要点为脾气虚弱无以运化水谷精微，使筋脉失养以致“脾无以主肌肉”而发病，且脾气虚弱日久见湿热、瘀血、痰浊等浊邪堆积。本项目前期研究[3]从“脾气虚弱兼浊邪堆积”的角度为切入点，认为本病多以虚实夹杂证为主，已证实葛根及其配伍不仅能改善肌无力症状[4-7]，且能作用于胃肠平滑肌线粒体[8]，具有较高的临床应用价值。本研究通过配制分别含有100 μg肽段的完全弗氏佐剂、不完全弗氏佐剂和磷酸盐缓冲液(PBS)的混合乳液诱导自身免疫性重症肌无力大鼠模型，观察葛根及其配伍对线粒体超微结构与肌肉生成调控因子[9]包括成纤维细胞生长因子(fibroblast growth factor，FGF)、神经生长因子(nerve growth factor，NGF)和胰岛素样生长因子(insulin-like growth factors， IGFs)的影响，旨在从促进肌肉生长调控因子的角度探讨复方配伍对抗肌无力的内在机制及主要起效配伍，进一步研究葛根及其配伍对MG的作用机制。, 百拇医药(李蒙 文颖娟 杨俊超 王江 柏鲁宁)
〔关键词〕 重症肌无力;葛根配伍;骨骼肌线粒体超微结构;肌肉生成调控因子
〔Abstract〕 Objective To explore the action mechanism of Radix Puerariae and its compatibility on the mitochondrial structure of skeletal muscle in myasthenia gravis rats. Methods (1)modeling： The model rats were established by injecting 200 μL (including r 97-116∶100 μg) immune emulsion into abdomen， foot pad and back subcutaneously. After 30 days and 45 days， the rats were inoculated with the same method， and Lennon score ≥ 1 was used as the model rats. (2)Grouping and treatment： In addition to the blank group， the successfully modeling rats were randomly divided into a model group， a prednisone group， a Radix Puerariae group， a Geqi group， a Geshen group， a Gemiao group and a Geqishenmiao group， with 20 rats in each group. After 2 weeks of successful modeling， the blank group and model group were given distilled water by gavage， and the other groups were given drugs by gavage for 4 weeks. (3)Sample detection： The mitochondrial ultrastructure of skeletal muscle was observed by transmission electron microscope. The expression of FGF， NGF and IGFs was detected by immunohistochemistry. Results Compared with the model group， the mitochondrial morphology of each group recovered， and the whole morphology of the Geqi group and the Geqishenmiao group recovered better after treatment; Compared with the model group， the positive expression rates of FGF， NGF and IGFs in each treatment group increased， and the difference was statistically significant (P<0.01). Conclusion Radix Puerariae and its compatibility may promote the recovery of mitochondrial structure by improving the positive expression of myogenic regulatory factors， so as to achieve the purpose of treating myasthenia gravis.
〔Keywords〕 myasthenia gravis; compatibility of Radix Puerariae; ultrastructure of skeletal muscle mitochondria; regulation factors of muscle formation
重癥肌无力(myasthenia gravis，MG)是一种难治性自身免疫性疾病，以眼肌或全身肌无力为主要表现[1]，到目前为止，其病理机制尚不十分清楚，病因以乙酰胆碱受体(acetylcholine receptor， AchR)受损为主，即神经肌肉接头处突触后膜中因AchR分子数量减少无法产生足够的终板电位而导致肌肉传递功能障碍[2]，最终产生肌无力的症状。中医学认为MG病机要点为脾气虚弱无以运化水谷精微，使筋脉失养以致“脾无以主肌肉”而发病，且脾气虚弱日久见湿热、瘀血、痰浊等浊邪堆积。本项目前期研究[3]从“脾气虚弱兼浊邪堆积”的角度为切入点，认为本病多以虚实夹杂证为主，已证实葛根及其配伍不仅能改善肌无力症状[4-7]，且能作用于胃肠平滑肌线粒体[8]，具有较高的临床应用价值。本研究通过配制分别含有100 μg肽段的完全弗氏佐剂、不完全弗氏佐剂和磷酸盐缓冲液(PBS)的混合乳液诱导自身免疫性重症肌无力大鼠模型，观察葛根及其配伍对线粒体超微结构与肌肉生成调控因子[9]包括成纤维细胞生长因子(fibroblast growth factor，FGF)、神经生长因子(nerve growth factor，NGF)和胰岛素样生长因子(insulin-like growth factors， IGFs)的影响，旨在从促进肌肉生长调控因子的角度探讨复方配伍对抗肌无力的内在机制及主要起效配伍，进一步研究葛根及其配伍对MG的作用机制。, 百拇医药(李蒙 文颖娟 杨俊超 王江 柏鲁宁)
参见：首页 > 医疗版 > 疾病专题 > 神经内科 > 肌病 > 重症肌无力