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[电针对脑缺血再灌注损伤大鼠一氧化氮合酶的影响]

[Effect of electroacupuncture on nitric oxide synthase in rats with cerebral ischemia-reperfusion injury].

作者信息

Chen Shi-xin, Ding Mao-chao, Dai Kai-yu

机构信息

Department of Anatomy, Wenzhou Medical College, Wenzhou 325035.

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2011 Jun;31(6):784-8.

Abstract

OBJECTIVE

To study the effect of electroacupuncture on nitric oxide synthase (NOS) in rats with cerebral ischemia-reperfusion injury.

METHODS

Focal cerebral ischemia-reperfusion model was established using modified intravascular suture technique. The NO content in the brain tissue was detected by nitrite reduction and the expressions of nNOS and iNOS were detected by immunohistochemistry. Eighty rats in this experiment were divided into the normal group, the cerebral ischemia-reperfusion injury model group (as the model group), the cerebral ischemia-reperfusion injury + electroacupuncture group (as the acupuncture group), and the cerebral ischemia-reperfusion injury + phosphatidylinositol 3 kinase (PI3-K) inhibitor group (as the inhibitor group). Each group consisted of twenty rats. Five microL PI3-K inhibitor LY294002 (400 microL) was slowly injected at the lateral cerebral ventricle of rats in the inhibitor group at a constant speed using microinjector according to Konig Klippel atlas of the stereotaxis instrument. Shuigou (DU26) and Chengjiang (RN24) were selected to determine levels of NO and NOS.

RESULTS

After 24-h ischemia-reperfusion, the NO levels of the hippocampus and the cerebral cortex increased abnormally, and the expressions of nNOS and iNOS increased, showing significant difference when compared with those of the normal group (P<0.05). By electroacupuncture at Shuigou (DU26) and Chengjiang (RN24), the ischemic cerebral ischemia-reperfusion injury neuron loss was inhibited. Meanwhile, the high levels of NO, nNOS and iNOS in the cerebral cortex and the hippocampus were significantly inhibited (P<0.05). The abnormally increased expressions of nNOS and iNOS were reversed, showing significant difference when compared with the model group (P<0.05). But when compared with the normal group, there was no significant difference (P>0.05). The effects of electroacupuncture reversed the abnormally increased NO levels of the hippocampus and the cerebral cortex and expressions of nNOS and iNOS after LY294002 oppressed anti-PI3K to block the TrkA acceptor circuit. The NO levels of the hippocampus and the cerebral cortex and expressions of nNOS and iNOS increased again, showing significant difference when compared with the acupuncture group (P<0.05).

CONCLUSIONS

Acupuncture fought against cerebral ischemia and reperfusion in the loss of neurons, at the same time, the abnormal regulation of NOS had reverse effect partly through TrkA/PI3K mediated signal transduction pathway.

摘要

目的

研究电针对脑缺血再灌注损伤大鼠一氧化氮合酶(NOS)的影响。

方法

采用改良血管内缝合技术建立局灶性脑缺血再灌注模型。采用亚硝酸盐还原法检测脑组织中NO含量,免疫组织化学法检测nNOS和iNOS的表达。本实验80只大鼠分为正常组、脑缺血再灌注损伤模型组(模型组)、脑缺血再灌注损伤+电针组(针刺组)、脑缺血再灌注损伤+磷脂酰肌醇3激酶(PI3-K)抑制剂组(抑制剂组)。每组20只。按照立体定向仪的Konig Klippel图谱,用微量注射器将5 μL PI3-K抑制剂LY294002(400 μL)以恒定速度缓慢注入抑制剂组大鼠侧脑室。选取水沟(DU26)和承浆(RN24)测定NO和NOS水平。

结果

缺血再灌注24小时后,海马和大脑皮质NO水平异常升高,nNOS和iNOS表达增加,与正常组比较差异有统计学意义(P<0.05)。针刺水沟(DU26)和承浆(RN24)抑制了缺血性脑缺血再灌注损伤神经元丢失。同时,大脑皮质和海马中高水平的NO、nNOS和iNOS被显著抑制(P<0.05)。nNOS和iNOS异常增加的表达得到逆转,与模型组比较差异有统计学意义(P<0.05)。但与正常组比较,差异无统计学意义(P>0.05)。LY294002抑制抗PI3K阻断TrkA受体通路后,电针的作用逆转了海马和大脑皮质中异常升高的NO水平以及nNOS和iNOS的表达。海马和大脑皮质的NO水平以及nNOS和iNOS的表达再次升高,与针刺组比较差异有统计学意义(P<0.05)。

结论

针刺对抗脑缺血再灌注所致的神经元丢失,同时,NOS的异常调节部分通过TrkA/PI3K介导的信号转导通路产生逆转作用。

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