针刺对幼鼠HIBD脑组织发育及NGF蛋白表达的实验研究(1)

The effect of acupuncture on the brain growth and expression of NGF in brain tissue of the HIBD newborn rats

目的探讨针刺对新生大鼠缺氧缺血性脑损伤(HIBD)脑组织重量及NGF表达的作用及其机制。方法选用85只7日龄清洁级SD大鼠,将大鼠结扎左侧颈总动脉后恢复2h,置于透明密闭容器中,并入37℃恒温水中以1L/min的速度通入低氧气体,2.5h后将动物取出,将存活者继续保温1h时后作行为测定,翻身不能、平衡异常或左旋者视为成功的模型。共72只,随机分为3组,A组为HIBD+针刺Ⅰ组,B组为HIBD+针刺Ⅱ组,C组为HIBD对照组。另设假手术组D组,每组24只。选用幼鼠百会、患侧颞Ⅰ针、内关、曲池、足三里、涌泉,A组于造模后24h开始针刺,前7d仅针四肢部穴位,第8d开始加针头部穴位,B组造模后第8d开始针刺,头、体针同步。针刺均为1次/d,A组连续针刺20d,B组连续针刺13d,假手术组干预措施与A组相同,至HIBD后第21d处死取材,测量各组脑组织重量,采用免疫组化方法测定针刺对HIBD大鼠海马区NGF表达,用图像分析仪在光镜下计算及NGF阳性表达的细胞数目。结果①动物模型的存活率、前肢触觉刺激试验和脑重量的变化结果,都是针刺组优于对照组(P<0.01);②4组左侧海马CA1区神经元NGF阳性数差异有统计学意义,其中D<C<B<A(P<0.001)。结论电针对HIBD确有保护作用,机制与其增强脑内NGF生物活性有关。

Objective To study the protective effect and the mechanism of acupuncture on hypoxia-- ischemic brain damage （HIBD） in newborn rats. Methods Eighty-five clean conventional SD rats, aged 7 days, were selected. The rats were operated by ligation of left common carotid artery and recovered after 2 hours. Then they were put in hyaline airtight vessel in the homeothermal water of 37℃. Hypoxia gas was infused to the vessel at the speed of 1L/min. The animal were took out after 2.5 hours. The successful models were evaluated by the act of inability of roll over, abnormal balance or turning left. Seventy-two successful HIBD models were randomly divided into 3 groups. Group A was HIBD＋ acupuncture Ⅰ . Group B was HIBD＋ acupuncture Ⅱ. In addition, Group C was control group of HIBD. Sham--operated group was included into group D. The number of the rats in each group was 24. The acupoints of Bailaui, Ⅰ needle of temple in the affected lateral, Neiguan, Quchi, Zusanli, Yongquan were selected. Group A were acupunctured at 24 hours after the models were made. Only four extremities were acupunctured in initial 7 days. The headacupoints were acupunctured in addition at 8th day. Group B began to be acupunctured at 8th day after the models made. Head and body acupuncture were operated at the same time. In both of the two grus. The acupuncture operated once a day. Group A continued 20 days. Group B continued 13 days. Sham--operated group was interfered as same as group A. The rats were put to death 21 days after the HIBD, the brain weight were measured and the expression of NGF in the hippocampus of HIBD rat were observed by im- munohistochemistry staining assay respectively. NGF positive cell numbers were counted by the equipment of picture analysis at the light microscope. The difference between the groups was analyzed by analysis of variance. Results The difference of expression of NGF in the CA1 zone of hippocampus of all the four groups has obvious significance in statistics, D 〈 C 〈 B〈 A （P〈 0.001） . Conclusion The results verify the protective efficacy of acupuncture on HIBD, which has relation to the enhancement of the expression of NGF in the brain.