肌力康饮对EAMG大鼠血清AchR-Ab及TGF-β1水平的影响(1)
摘 要:目的:观察中药复方肌力康饮对实验性自身免疫性重症肌无力(EAMG)大鼠血清AchR-Ab、TGF-β1水平的影响,以探讨其治疗重症肌无力的可能机制。方法:60只Lewis大鼠随机分为6组,每组10只:空白组,模型组,阳性药物组,肌力康饮高、中、低剂量组。采用AchR主动免疫的方法免疫除空白组外的各组大鼠,制备EAMG大鼠模型。确立模型成功,各组给药治疗4周后,采用ELISA法检测血清AchR-Ab、TGF-β1水平。结果:肌力康饮各组、阳性药物组能明显改善EAMG大鼠肌无力症状,临床症状评分显著降低、血清AchR-Ab水平明显下降、TGF-β1水平明显升高,与模型组比较,有显著性差异(P<0.01);肌力康饮高剂量组与阳性药物组比较亦有明显差异(P<0.05);但肌力康饮各剂量组之间比较无显著性差异(P>0.05)。结论:肌力康饮下调了EAMG大鼠过高的AchR-Ab水平,并使TGF-β1升高,从而改善了EAMG大鼠的临床症状。
, 百拇医药 关键词:肌力康饮;EAMG;TGF-β1;AchR-Ab
中图分类号:R285.5 文献标识码:A 文章编号:1673-7717(2010)01-0080-03
Effect of Ji Li Kang Drink on Eoperimental Autoimmune Myasthenia Gravis Serum
AchR-Ab and TGF-β1 Levels
NIU Fengyun,CHEN Huijuan,JIA Weigang,ZHAO Lili
(Heilongjiang Academy of Traditional Chinese Medicine, Harbin 150036, Heilongjiang, China)
, 百拇医药
Abstract:Objective: Observe the effect of Ji Li Kang Drink on experimental autoimmune myasthenia gravis (EAMG) serum AchR-Ab, TGF-β1 levels in order to explore the possible mechanisms of treatment of myasthenia gravis.Methods: 60 Lewis rats were randomly divided into six groups of 10: blank group, model group, positive medicine group, high-dose group of Ji Li Kang drink ,medium-dose group and low-dose group. Method of active immunization with AchR was used to prepare for EAMG rats. After being established a successful model, each group were detected serum AchR-Ab, TGF-β1 levels using ELISA after 4 weeks drug treatment.Results: groups of Ji Li Kang drink and positive medicine group can significantly improve the symptoms of EAMG rats;Score of clinical symptom significantly decreased ; AchR-Ab levels in serum were significantly decreased;TGF-β1 levels were significantly elevated compared with model group.There was significant difference (P<0.01). Groups of Ji Li Kang drink and the positive medicine group also were significantly different (P<0.05);Each dose group of Ji Li Kang drink showed no significant difference (P>0.05). Conclusion: Ji Li Kang drink reduced AchR-Ab level , and increased TGF-β1 level of EAMG rats Thus Ji Li Kang drink improved the clinical symptoms ofEAMGrats.
, 百拇医药
Key words:Ji Li Kang Decoction; EAMG;TGF-β1; AchR-Ab
重症肌无力(myasthenia gravis,MG)主要是由乙酰胆碱受体抗体介导的,补体参与、细胞免疫依赖的,针对神经肌肉接头处突触后膜上乙酰胆碱受体的自身免疫性疾病[1]。目前西医治疗多采用抗胆碱酯酶药、免疫抑制剂、糖皮质激素等药物,副作用大,依赖性高,复发率高,远期疗效差。基于对中医药治疗该病多年经验的总结,肌力康饮从健脾益损立法,用黄芪、党参、白术等药治疗重症肌无力,疗效显著。本实验通过建立EAMG大鼠模型,观察大鼠血清AchR-Ab、TGF-β1的变化,探讨肌力康饮抗重症肌无力的作用机制。
1材料与方法
1.1动物
Lewis大鼠60只,雌性,体重160~180g,购自中国医学科学院动物研究所,合格证号:SCXK(京) 2005-0013。
, 百拇医药
1.2 药品与试剂
肌力康饮,黑龙江省中医研究院制剂室提供,规格:10mL/支,批号:20080401。溴新斯的明片,由杭州沿山化工有限公司生产,15mg×60片/瓶,碾碎溶于蒸馏水中使用。Acetylcholine Receptorα1,Fragment 129-145,购自Sigma, 500μg/支,货号A2352,纯度≥93%。完全福氏佐剂(CFA),购自Sigma,10mL/支,货号F-5881。AchR-Ab酶联试剂盒,96孔,购自博润生物有限公司;TGF-β1酶联试剂盒,96孔,购自博润生物有限公司。
1.3 造模
1.3.1 抗原乳化
用PBS缓冲液将Acetylcholine Receptor α1,Fragment 129-145溶解,待用;将等量的CFA用注射器抽取装入无菌小瓶(内置打磨光滑的自制转子),置于磁力搅拌器上,将PBS液用注射器逐渐加入到小瓶中,旋转3～5min后,呈乳白色较黏稠液体,抽取一滴滴入清水中不分散而呈现球形为乳化成功。, http://www.100md.com(牛凤云 陈会娟 贾维刚 赵丽丽)
, 百拇医药 关键词:肌力康饮;EAMG;TGF-β1;AchR-Ab
中图分类号:R285.5 文献标识码:A 文章编号:1673-7717(2010)01-0080-03
Effect of Ji Li Kang Drink on Eoperimental Autoimmune Myasthenia Gravis Serum
AchR-Ab and TGF-β1 Levels
NIU Fengyun,CHEN Huijuan,JIA Weigang,ZHAO Lili
(Heilongjiang Academy of Traditional Chinese Medicine, Harbin 150036, Heilongjiang, China)
, 百拇医药
Abstract:Objective: Observe the effect of Ji Li Kang Drink on experimental autoimmune myasthenia gravis (EAMG) serum AchR-Ab, TGF-β1 levels in order to explore the possible mechanisms of treatment of myasthenia gravis.Methods: 60 Lewis rats were randomly divided into six groups of 10: blank group, model group, positive medicine group, high-dose group of Ji Li Kang drink ,medium-dose group and low-dose group. Method of active immunization with AchR was used to prepare for EAMG rats. After being established a successful model, each group were detected serum AchR-Ab, TGF-β1 levels using ELISA after 4 weeks drug treatment.Results: groups of Ji Li Kang drink and positive medicine group can significantly improve the symptoms of EAMG rats;Score of clinical symptom significantly decreased ; AchR-Ab levels in serum were significantly decreased;TGF-β1 levels were significantly elevated compared with model group.There was significant difference (P<0.01). Groups of Ji Li Kang drink and the positive medicine group also were significantly different (P<0.05);Each dose group of Ji Li Kang drink showed no significant difference (P>0.05). Conclusion: Ji Li Kang drink reduced AchR-Ab level , and increased TGF-β1 level of EAMG rats Thus Ji Li Kang drink improved the clinical symptoms ofEAMGrats.
, 百拇医药
Key words:Ji Li Kang Decoction; EAMG;TGF-β1; AchR-Ab
重症肌无力(myasthenia gravis,MG)主要是由乙酰胆碱受体抗体介导的,补体参与、细胞免疫依赖的,针对神经肌肉接头处突触后膜上乙酰胆碱受体的自身免疫性疾病[1]。目前西医治疗多采用抗胆碱酯酶药、免疫抑制剂、糖皮质激素等药物,副作用大,依赖性高,复发率高,远期疗效差。基于对中医药治疗该病多年经验的总结,肌力康饮从健脾益损立法,用黄芪、党参、白术等药治疗重症肌无力,疗效显著。本实验通过建立EAMG大鼠模型,观察大鼠血清AchR-Ab、TGF-β1的变化,探讨肌力康饮抗重症肌无力的作用机制。
1材料与方法
1.1动物
Lewis大鼠60只,雌性,体重160~180g,购自中国医学科学院动物研究所,合格证号:SCXK(京) 2005-0013。
, 百拇医药
1.2 药品与试剂
肌力康饮,黑龙江省中医研究院制剂室提供,规格:10mL/支,批号:20080401。溴新斯的明片,由杭州沿山化工有限公司生产,15mg×60片/瓶,碾碎溶于蒸馏水中使用。Acetylcholine Receptorα1,Fragment 129-145,购自Sigma, 500μg/支,货号A2352,纯度≥93%。完全福氏佐剂(CFA),购自Sigma,10mL/支,货号F-5881。AchR-Ab酶联试剂盒,96孔,购自博润生物有限公司;TGF-β1酶联试剂盒,96孔,购自博润生物有限公司。
1.3 造模
1.3.1 抗原乳化
用PBS缓冲液将Acetylcholine Receptor α1,Fragment 129-145溶解,待用;将等量的CFA用注射器抽取装入无菌小瓶(内置打磨光滑的自制转子),置于磁力搅拌器上,将PBS液用注射器逐渐加入到小瓶中,旋转3～5min后,呈乳白色较黏稠液体,抽取一滴滴入清水中不分散而呈现球形为乳化成功。, http://www.100md.com(牛凤云 陈会娟 贾维刚 赵丽丽)