针刺不同时间干预对TBI模型大鼠BDNF蛋白表达及mRNA水平调控的实验研究

Study on the Regulating BDNF and mRNA of Neuron with TBI Rats Model in Different Periods

目的研究不同时间点针刺介入治疗TBI的效果,揭示针刺干预对TBI动物神经元的保护作用及部分机理,为临床针刺治疗TBI的最佳介入时间提供实验依据。方法将SD大鼠随机分为空白组、针刺1组、针刺2组、针刺3组和模型组,使用eCCI仪器制备TBI动物模型。取百会、人中、内关(左侧)和足三里(左侧),分别在脑损伤后第1日、第3日、第5日开始针刺治疗,各组治疗持续10次,在第15日取材,观察、检测相关指标:免疫组化法、Western-blot法检查脑组织中BDNF蛋白的定位、含量及荧光定量PCR法检测mRNA的表达。结果采用免疫组化检测脑组织中BDNF蛋白表达定位及含量,又结合Western-blot定量检测其蛋白含量变化,针刺可上调脑组织损伤皮质中NGF蛋白的含量;同时,荧光定量PCR检测也发现,针刺可上调脑组织损伤皮质中BDNF相应基因mRNA含量,且针刺1组优于针刺2组、针刺3组,组间比较具有统计学意义(P<0.01)。结论 TBI后针刺介入治疗的时间点应尽可能早,更有利于发挥针刺上调脑组织神经元的神经生长因子BDNF及mRNA表达,达到保护脑组织神经元的作用。

Objective To prepare TBI model of SD rat by the method, and study the effects of acupuncture treatment of TB1 at different periods. To reveal protective effects and mechanism of the acupuncture to the injured nerve cells of TBI brain,provide experimental basis of acupuncture treatment of acute phase TBI at the best time,and to add the scientific contents of acupuncture treatment of TBI. Methods 70 SD rats were randomly divided into blank control group,acupuncture group 1 ,acupuncture group 2,acupuncture group 3 and model control group. The TBI animal models were prepared with the eCCI of the speed of 4m/s and the depth of 3ram. Than rats were treated by aeupuneturing Baihui （DU20）, Renzhong（ DU26 ）, the left Neiguan （PC6） and the left Zusanli（ST36） ） after the brain injured at 1 day,3 days,5 days. The treatments of each group were repeated 10 times. The materials were collected at the 15th day and to observe and examine the related index signs：By immunohistochemical and Western -blot to examine the localization and content of BDNF in the brain tissue protein, to detect the mRNA changes of BDNF by Fluorescence metered PCR method. Results The expression and content of BDNF were detected by immunohistochemistry and Western - blot in brain tissue. Acupuncture can upregulate NGF. And by PCR, acupuncture can increase the mRNA of BDNF. After TBI occurred,while acupuncture treatment was used as soon as possible,the acupuncture was more obvious on the regulation effect. And there was significant differences among the three groups （P 〈O. 01 ）. Conclusion Acupuncture treated TBI as early as possible. The results of this study showed that after 1 day of TBI occurred if acupuncture was used immediately, the curative effect was better than 3 days and 5 days latter.