头针电刺激对急性脑梗死大鼠CD40、CD40L的影响
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摘要
目的研究头针电刺激对急性脑梗死大鼠脑组织中CD40、CD40L含量的影响。方法将96只健康的雄性Wistar大鼠随机分为头针电刺激组、假手术组和模型对照组,每组又根据治疗的时间点不同分为4个亚组(1d组、3d组、7d组和14d组),每组8只大鼠。头针电刺激组及模型对照组采用改良的线栓法制作右侧大脑中动脉闭塞局灶性脑缺血大鼠模型,3组大鼠在造模成功后第1天起,头针电刺激组给予大鼠针刺干预,取百会穴及大椎穴,以疏密波进行电刺激治疗;假手术组及模型对照组大鼠在同一时间固定于针刺台,不做处理。于治疗前及治疗后第1天、第3天、第7天和第14天,检测脑组织中CD40、CD40L的表达。3组大鼠造模结束清醒后及治疗前后均进行Bederson评分。结果治疗后第7天、第14天,头针电刺激组较模型对照组Bederson评分明显降低(P<0.05);脑组织中CD40、CD40L的表达均明显降低(P<0.01)。其中头针电刺激组中治疗后14d脑组织中CD40、CD40L的表达、Bederson评分较治疗前明显降低,差异有统计学意义(P<0.05)。结论头针电刺激能显著改善急性脑梗死大鼠神经功能,从而改善肢体运动功能,其作用机制可能是抑制CD40、CD40L的表达,从而下调其介导的炎症及免疫分子生物学因素有关,且以治疗14d效果最优。
Objective To study the effects of scalp electroacupuncture on CD40 and CD40L in brain tissue of rats with acute cerebral infarction.Methods Ninety-six healthy male wistar rats were randomly divided into scalp electroacupuncture group,sham operation group and model control group.The rats in each group were divided into 4 subgroups:1-day group,3-days group,7-day group and 14-day group according to the treatment time,8 rats in each group.The rat model of focal cerebral ischemia with right middle cerebral artery occlusion was established by modified suture method in scalp electroacupuncture group and model control group.After one day of successful modeling,the rats were treated with scalp electroacupuncture on Baihui acupoint(GV20)and Dazhui acupoint(DU14)in scalp electroacupuncture group.No treatment was required in sham operation group and model control group.The expression of CD40 and CD40L in brain tissue was detected before treatment,1 day,3 days,7 days and14 days after treatment.Bederson score was evaluated after awake and before and after treatment.Results After 7 days and 14 days of treatment,Bederson scores decreased(P <0.05),the neurological functions were significantly improved,the expression of CD40 and CD40L in brain tissue was reduced in scalp electroacupuncture group compared with model control group(P <0.01).The expression of CD40 and CD40L in the 14-day subgroup was significantly lower than that in the pre-treatment group compared with the sham-operated group(P <0.05),and the Bederson score was significantly lower(P <0.05).Conclusion Scalp electroacupuncture can significantly improve the neurological function in rats with acute cerebral infarction,thereby improving limb motor function.The mechanism may be related to the inhibition of CD40 and CD40L expressions and the down-regulation of inflammatory and immune molecular biology factors.Treatment for 14 days is the best therapy.
Objective To study the effects of scalp electroacupuncture on CD40 and CD40L in brain tissue of rats with acute cerebral infarction.Methods Ninety-six healthy male wistar rats were randomly divided into scalp electroacupuncture group,sham operation group and model control group.The rats in each group were divided into 4 subgroups:1-day group,3-days group,7-day group and 14-day group according to the treatment time,8 rats in each group.The rat model of focal cerebral ischemia with right middle cerebral artery occlusion was established by modified suture method in scalp electroacupuncture group and model control group.After one day of successful modeling,the rats were treated with scalp electroacupuncture on Baihui acupoint(GV20)and Dazhui acupoint(DU14)in scalp electroacupuncture group.No treatment was required in sham operation group and model control group.The expression of CD40 and CD40L in brain tissue was detected before treatment,1 day,3 days,7 days and14 days after treatment.Bederson score was evaluated after awake and before and after treatment.Results After 7 days and 14 days of treatment,Bederson scores decreased(P <0.05),the neurological functions were significantly improved,the expression of CD40 and CD40L in brain tissue was reduced in scalp electroacupuncture group compared with model control group(P <0.01).The expression of CD40 and CD40L in the 14-day subgroup was significantly lower than that in the pre-treatment group compared with the sham-operated group(P <0.05),and the Bederson score was significantly lower(P <0.05).Conclusion Scalp electroacupuncture can significantly improve the neurological function in rats with acute cerebral infarction,thereby improving limb motor function.The mechanism may be related to the inhibition of CD40 and CD40L expressions and the down-regulation of inflammatory and immune molecular biology factors.Treatment for 14 days is the best therapy.
引文
[1]张楠楠,陈会生.依达拉奉对脑梗死大鼠神经保护作用最佳治疗时间窗的探讨[J].解放军医药杂志,2016,28(3):18-21.
[2]李楠,白宏英.脑梗死大鼠CD4+、CD25+调节性T细胞的作用[J].中国实用神经疾病杂志,2015,18(20):45-46.
[3]李晓宁,迟蕾,吴磊,等.头针对急性脑梗死大鼠不同时间窗Bax与Bcl-2影响的研究[J].针灸临床杂志,2016,21(4):73-76;97.
[4]谢惠芳,徐如祥,陈忠灿.线栓法制作大鼠局灶性缺血再灌注损伤模型的改进[J].中华神经医学杂志,2007,6(4):340-342.
[5]唐中华,王强,封海涛,等.Ⅱ型大麻受体激动剂JWH133对大鼠局灶性脑缺血-再灌注损伤的神经保护作用[J].中国药业,2016,25(18):21-24.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:327-329.
[7]Bederson JB,Pitts LH,Germano SM,et al.Evaluation of 2,3,5-triphenyl tetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats[J].Stroke,1986,17(6):1304-1308.
[8]李涤生.临床检验基础[M].北京:人民卫生出版社,1991:91-112.
[9]孙寒静,刘志和,吴艳华,等.川芎嗪对急性脑梗死大鼠CD40/CD40L信号通路的影响及机制探讨[J].中西医结合心脑血管病杂志,2016,14(10):1087-1090.
[10]艾宗耀,易燕锋,肖梅红,等.活血通络解毒方对脑缺血Wistar大鼠血液流变学的影响[J].中国中医急症,2016,25(7):1288-1289;1322.
[11]Mitsios N,Gaffney J,Kumar P,et al.Pathophysiology of acute ischaemic stroke:an analysis of common signalling mechanisms and identification of new molecular targets[J].Pathobiology,2006,73(4):159-175.
[12]Kaushal V,Schlichter LC.Mechanisms of microglia mediated neurotoxicity in a new model of the stroke penumbra[J].Journal of Neuroscience,2008,28(9):2221-2230.
[13]梁晓,张允岭,王新祥,等.清热活血组分对急性脑梗死火毒症大鼠NF-κB炎症信号通路调控作用研究[J].北京中医药大学学报,2015,38(6):377-382.
[14]Pinelli DF,Ford ML.Novel insights into anti-CD40/CD154immunotherapy in transplant tolerance[J].Immunotherapy,2015,7(4):399-410.
[15]Okwor I,Jia P,Uzonna JE,et al.Interaction of macrophage antigen 1and CD40ligand leads to IL-12production and resistance in CD40-deficient mice infected with leishmania major[J].Journal of Immunology,2015,195(7):3218-3226.
[16]董红锰,白纯,李先亮,等.CD40-CD40L信号通路在移植免疫中的应用与进展[J].器官移植,2016,7(4):308-311.
[17]李晓宁,王倩,刘双岭,等.头针治疗急性脑梗死大鼠不同时间窗的疗效观察[J].针灸临床杂志,2014,30(11):57-59.
[18]张纯,刘健,林秋虹,等.电针治疗急性脑梗死疗效观察[J].现代诊断与治疗,2013,24(13):2913-2914.
[19]武廷格,宿宝贵,吕来清,等.电针下调脑梗死大鼠皮质缺血半影区Bax蛋白质表达[J].中华临床医师杂志,2009,3(2):222-228.
[2]李楠,白宏英.脑梗死大鼠CD4+、CD25+调节性T细胞的作用[J].中国实用神经疾病杂志,2015,18(20):45-46.
[3]李晓宁,迟蕾,吴磊,等.头针对急性脑梗死大鼠不同时间窗Bax与Bcl-2影响的研究[J].针灸临床杂志,2016,21(4):73-76;97.
[4]谢惠芳,徐如祥,陈忠灿.线栓法制作大鼠局灶性缺血再灌注损伤模型的改进[J].中华神经医学杂志,2007,6(4):340-342.
[5]唐中华,王强,封海涛,等.Ⅱ型大麻受体激动剂JWH133对大鼠局灶性脑缺血-再灌注损伤的神经保护作用[J].中国药业,2016,25(18):21-24.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:327-329.
[7]Bederson JB,Pitts LH,Germano SM,et al.Evaluation of 2,3,5-triphenyl tetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats[J].Stroke,1986,17(6):1304-1308.
[8]李涤生.临床检验基础[M].北京:人民卫生出版社,1991:91-112.
[9]孙寒静,刘志和,吴艳华,等.川芎嗪对急性脑梗死大鼠CD40/CD40L信号通路的影响及机制探讨[J].中西医结合心脑血管病杂志,2016,14(10):1087-1090.
[10]艾宗耀,易燕锋,肖梅红,等.活血通络解毒方对脑缺血Wistar大鼠血液流变学的影响[J].中国中医急症,2016,25(7):1288-1289;1322.
[11]Mitsios N,Gaffney J,Kumar P,et al.Pathophysiology of acute ischaemic stroke:an analysis of common signalling mechanisms and identification of new molecular targets[J].Pathobiology,2006,73(4):159-175.
[12]Kaushal V,Schlichter LC.Mechanisms of microglia mediated neurotoxicity in a new model of the stroke penumbra[J].Journal of Neuroscience,2008,28(9):2221-2230.
[13]梁晓,张允岭,王新祥,等.清热活血组分对急性脑梗死火毒症大鼠NF-κB炎症信号通路调控作用研究[J].北京中医药大学学报,2015,38(6):377-382.
[14]Pinelli DF,Ford ML.Novel insights into anti-CD40/CD154immunotherapy in transplant tolerance[J].Immunotherapy,2015,7(4):399-410.
[15]Okwor I,Jia P,Uzonna JE,et al.Interaction of macrophage antigen 1and CD40ligand leads to IL-12production and resistance in CD40-deficient mice infected with leishmania major[J].Journal of Immunology,2015,195(7):3218-3226.
[16]董红锰,白纯,李先亮,等.CD40-CD40L信号通路在移植免疫中的应用与进展[J].器官移植,2016,7(4):308-311.
[17]李晓宁,王倩,刘双岭,等.头针治疗急性脑梗死大鼠不同时间窗的疗效观察[J].针灸临床杂志,2014,30(11):57-59.
[18]张纯,刘健,林秋虹,等.电针治疗急性脑梗死疗效观察[J].现代诊断与治疗,2013,24(13):2913-2914.
[19]武廷格,宿宝贵,吕来清,等.电针下调脑梗死大鼠皮质缺血半影区Bax蛋白质表达[J].中华临床医师杂志,2009,3(2):222-228.