脑复合剂对创伤性脑损伤模型大鼠脑内NO、nNOS活性及表达的影响

中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (12): 1340-1346.

脑复合剂对创伤性脑损伤模型大鼠脑内NO、nNOS活性及表达的影响

苗静琨, 陈启雄, 吴小玫, 刘振球, 张晓萍

重庆医科大学附属儿童医院新生儿科,儿童发育疾病研究省部共建教育部重点实验室,儿科学重庆市重点实验室,重庆市儿童发育重大疾病诊治与预防国际科技合作基地,重庆 400014

收稿日期:2011-09-13 修回日期:2011-11-18 出版日期:2011-12-26 发布日期:2012-01-07
通讯作者: 陈启雄,通信作者,男,博士,研究方向:脑损伤发病机制及防治。Tel: 13193039866 E-mail: chengqixiong@126.com
作者简介:苗静琨,女,硕士,研究方向:脑损伤的发病机制及防治。Tel: 13110207118 E-mail: jennamiao@yahoo.com.cn
基金资助:
重庆市卫生局中医药科研资助项目(渝中医20024270)

Effects of compound decotion on activity and expression of nitric oxide and neuronal nitric oxide synthase in the brain of traumatic brain injury rat model

MIAO Jing-kun, CHEN Qi-xiong, WU Xiao-mei, LIU Zhen-qiu, ZHANG Xiao-ping

Department of Neonatology, the Children's Hospital, Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing 400014, China

Received:2011-09-13 Revised:2011-11-18 Online:2011-12-26 Published:2012-01-07

摘要/Abstract

摘要： 目的: 探讨脑复合剂治疗对创伤性脑损伤(traumatic brain injury, TBI)模型大鼠脑保护作用可能的分子机制。方法: 用自由落体打击(Feeney法)建立TBI模型,分别设立假手术组、TBI模型组及中药治疗组。中药治疗组给予脑复合剂 10 g·kg^-1·d^-1,假手术组及TBI模型组给予同等剂量的等渗盐水,2 次/d,连续　7 d。分别于TBI后 24 h、72 h、1周等3个时相处死大鼠,用HE染色法观察大脑皮层的形态学变化,化学法检测NO、一氧化氮合酶(NOS)含量、神经元型NOS(nNOS)活性,免疫组化检测nNOS蛋白表达,原位杂交检测nNOS-mRNA表达。结果: TBI模型组各时相大鼠脑内NO、NOS含量、nNOS活性及表达、nNOS-mRNA表达均有不同程度增高,于TBI后 24 h 增高最为明显,持续至1周仍高于假手术组。而中药治疗组各时相大鼠脑组织NO、NOS含量、nNOS活性及表达、nNOS-mRNA表达均明显低于TBI模型组(P＜0.05),并于伤后1周接近正常水平。结论: 脑复合剂对TBI的保护作用可能与抑制TBI后nNOS-mRNA表达,降低脑组织nNOS活性,从而抑制NO的异常增高,减轻NO介导的神经细胞毒性有关。

关键词: 创伤性脑损伤, 一氧化氮, 神经元型一氧化氮合酶

Abstract: AIM: To explore the possible mechanism of the neuroprotective effect of compound decotion on traumatic brain injury model.METHODS: The traumatic brain injury rat model was induced by improved Feeneys fall weight method and divided into sham operation group, TBI model group and compound decotion treatment group, compound decotion treatment group rats were intragastrically administered with compound decotion at 10 g·kg^-1·d^-1. Sham operation group and TBI model group were treated with same dose of normal saline, twice per day for seven days. The brains were taken out at 24, 72 h and one week after injury, respectively. The morphological changes were observed by HE staining in the cerebral cortex. The NO and NOS concentration and neuronal nitric oxide synthase (nNOS) activity were detected by neurochemistry stain method. nNOS expression were detected by immunohistochemical method. nNOS-mRNA expression were detected by in situ hybridization.RESULTS: In TBI model group, the concentration of NO and NOS and the activity and expression of nNOS and the expression of nNOS- mRNA were markedly elevated at different degrees at different time point and especially 24 hours after injury and still higher than those of sham operation group one week after injury. However, compared with the TBI model group, the concentration of NO and NOS and the activity and expression of nNOS and the expression of nNOS-mRNA in compound decotion treatment group were obviously decreased and reach normal level one week after injury (P＜0.05).CONCLUSION: NO and nNOS play an important role in the pathophysiological process of TBI. Inhibition of activated NO and nNOS and mitigate secondary brain injury may represent a potential neuroprotective mechanism for the treatment of TBI with compound decotion.

Key words: Traumatic brain injury, Nitric oxide, Neuronal nitric oxide synthase

中图分类号:

R965.2

苗静琨, 陈启雄, 吴小玫, 刘振球, 张晓萍. 脑复合剂对创伤性脑损伤模型大鼠脑内NO、nNOS活性及表达的影响[J]. 中国临床药理学与治疗学, 2011, 16(12): 1340-1346.

MIAO Jing-kun, CHEN Qi-xiong, WU Xiao-mei, LIU Zhen-qiu, ZHANG Xiao-ping. Effects of compound decotion on activity and expression of nitric oxide and neuronal nitric oxide synthase in the brain of traumatic brain injury rat model[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(12): 1340-1346.

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