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针刺联合大鼠神经生长因子对脑瘫幼鼠神经行为能力及其脑组织生长与代谢相关蛋白的研究

[Study of acupuncture combined with rat nerve growth factor on neurobehavioral ability of cerebral palsy infant rats and its brain tissue growth and metabolism associated proteins].

作者信息

Zhao Pengju, Li Enyao

机构信息

Department of Children Rehabilitation Medicine, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China.

出版信息

Zhongguo Zhen Jiu. 2018 Jun 12;38(6):631-6. doi: 10.13703/j.0255-2930.2018.06.018.

Abstract

OBJECTIVE

To study the effects of acupuncture combined with rat nerve growth factor (NGF) on the cerebral palsy infant rats and the proteins which associated with growth, apoptosis and metabolism.

METHODS

Seventy infant rats were selected, Fifty infant rats of which were made the cerebral palsy infant model by the ligation of unilateral carotid artery for cerebral ischemia and oxygen-deficient environment, then the 30 model rats were randomly divided into a model group, a NGF group and a combined group, 10 rats in each group. Twenty infant rats were used in the sham-operated group and the blank control group, 10 rats in each group. The treatment was not given in the blank control group. The rats in the sham-operated group were cut the neck skin and separated the left carotid artery, and then sutured and disinfected the wound. The intraperitoneal injection of NGF (2000 U•kg•d) was used in the NGF group. Based on the injection in the NGF group, acupuncture was used in the combined group, once a day, and the acupoints were "Baihui" (GV 20), left "nieⅠ" (extra), "Dazhui" (GV 14), "Jizhong" (extra), "Quchi" (LI 11), "Yongquan" (KI 1), "Hegu" (LI 4), "Zhoujie" (extra) and "Xiqianxue" (extra). The same volume of saline was intraperitoneally injected in the model group for continuous 14 days. Neurobehavioral ability score was evaluated after treatment. TUNEL were conducted to detect the brain cell apoptosis rate and the expressions of apoptosis associated gene Bax, Bcl-2 and Casp3 were detected by PCR. The level of nerve growth associated protein (GAP-43) and energy metabolism-related protein monocarboxylate transporter protien 1(MCT 1) were detected by Western blot.

RESULTS

After intervention, the neurobehavioral ability of baby rats in the blank control and sham-operated group was normal, but there was various degrees of abnormity in the model group, NGF group and combined group. The scores of neurobehavioral ability of the combined group and NGF group were better than those of the model control (all <0.05), and the scores in the combined group was better than those in the NGF group (all <0.05). The left brain cell apoptosis rate, expressions of Bax and Casp3 in the combined group and NGF group were lower and the expressions of Bcl-2 were higher than those of the model group (all <0.05), with more obvious results of Bax and Gasp3 in the combined group than those in the NGF group (all <0.05). The protein levels of GAP-43 and MCT 1 in the combined group and NGF group were higher than those in the model group (all <0.05), with higher expressions in the combined group compared with those in the NGF group (both <0.05).

CONCLUSION

Acupuncture combined with NGF could improve the neurobehavioral ability of cerebral palsy infant rats, inhibit the nerve cell apoptosis and improve the brain tissue injure and energy metabolism by up-regulating the expressions of GAP-43 and MCT 1.

摘要

目的

研究针刺联合大鼠神经生长因子(NGF)对脑瘫幼鼠及与生长、凋亡和代谢相关蛋白质的影响。

方法

选取70只幼鼠,其中50只通过结扎单侧颈动脉造成脑缺血缺氧环境制备脑瘫幼鼠模型,将30只模型大鼠随机分为模型组、NGF组和联合组,每组10只。假手术组和空白对照组各20只幼鼠,每组10只。空白对照组不给予处理。假手术组切开颈部皮肤,分离左侧颈动脉,然后缝合伤口并消毒。NGF组腹腔注射NGF(2000 U•kg•d)。联合组在NGF组注射基础上给予针刺治疗,每日1次,穴位为“百会”(GV 20)、左侧“颞Ⅰ”(奇穴)、“大椎”(GV 14)、“脊中”(奇穴)、“曲池”(LI 11)、“涌泉”(KI 1)、“合谷”(LI 4)、“周节”(奇穴)和“膝前穴”(奇穴)。模型组腹腔注射等体积生理盐水,连续14天。治疗后评估神经行为能力评分。采用TUNEL法检测脑细胞凋亡率,并通过PCR检测凋亡相关基因Bax、Bcl-2和Casp3的表达。采用Western blot检测神经生长相关蛋白(GAP-43)和能量代谢相关蛋白单羧酸转运蛋白1(MCT 1)的水平。

结果

干预后,空白对照组和假手术组幼鼠神经行为能力正常,而模型组、NGF组和联合组存在不同程度异常。联合组和NGF组神经行为能力评分均优于模型对照组(均P<0.05),且联合组评分优于NGF组(均P<0.05)。联合组和NGF组左侧脑细胞凋亡率、Bax和Casp3表达低于模型组,Bcl-2表达高于模型组(均P<0.05),联合组Bax和Gasp3结果比NGF组更明显(均P<0.05)。联合组和NGF组GAP-43和MCT 1蛋白水平高于模型组(均P<0.05),联合组表达高于NGF组(均P<0.05)。

结论

针刺联合NGF可提高脑瘫幼鼠神经行为能力,抑制神经细胞凋亡,上调GAP-43和MCT 1表达改善脑组织损伤和能量代谢。

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