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[褪黑素在电针抗大鼠脑缺血再灌注损伤氧应激调节作用中的参与]

[Involvement of melatonin in the adjusting effect of electroacupuncture in resisting oxygen stress in cerebral ischemia-reperfusion injury rats].

作者信息

Li Zhong-Ren, Shen Mei-Hong, Niu Wen-Min

机构信息

Second Clinical Medical College of Nanjing University of Chinese Medicine, Nanjing 210029, China.

出版信息

Zhen Ci Yan Jiu. 2008 Jun;33(3):164-8.

Abstract

OBJECTIVE

To explore the effect of electroacupuncture (EA) in resisting acute cerebral ischemia-reperfusion injury (CI-RI) via anti-oxidation of melatonin (MT).

METHODS

A total of 52 male SD rats were randomized into Sham-operation (sham, n=8), CI-RI (model, n=8), EA (n=8), MT (3.2 mg/Kg, i.p., n=8), EA+MT (n=8), EA+Luzindole (L, a blocker of MT, 1 mg/100 microl/rat, n=6), EA+PD 98059 (a blocker of mitogen-activated protein kinase, MAPK, 1 mg 100 microl/rat, n=6) groups. EA (3 Hz,1-3 mA, continuous waves) was applied to "Dazhui" (GV 14) and "Baihui" (GV 20) for 30 min. CI-RI model was induced by right middle cerebral artery occlusion (MCAO) for 60 min and reperfusion for 24 h except sham-operation group. MT content of pineal gland tissue was determined by high performance liquid chromatography (HPLC) and neuroethological scores of the rats were evaluated by Kuluz's and Julio's methods. Bax and Bcl-2 gene protein expression of the striate body was detected by immunohistochemistry.

RESULTS

In comparison with sham group, the number of Bax immunoreactive (IR) positive cells and Bax/Bcl-2 in striate body increased significantly (P<0.001), and neuroethological score and Bcl-2 IR-positive cells decreased considerably in model group (P<0.01). Compared with model group, pineal MT content, neuroethological score and Bcl-2 IR-positive cells in EA group, neuroethological scores in EA+L and EA+P groups, and Bcl-2 expression in MT and EA+MT groups increased considerably (P<0.01, 0.001); while Bax IR-positive cells and Bax/Bcl-2 in EA, MT and EA+MT groups decreased very significantly (P<0.001). It showed that EA could obviously increase pineal MT content, improve CI-RI rats' activity score, upregulate Bcl-2 expression and down-regulate Bax expression in striate body. In comparison with simple EA group, neuroethological scores of EA+L and EA+P groups, Bax IR-positive cells and Bax/Bcl-2 of EA+MT group were significant lower (P<0.05, 0.01). No significant differences were found between EA+L and EA+P groups in behavior scores, and between MT and EA+MT groups in Bax and Bcl-2 IR-positive cell number and Bax/Bcl-2 (P>0.05).

CONCLUSION

EA has a good effect of anti-oxygen stress, protecting the brain from ischemic damage directly or indirectly, which may be related to its effect in upregulating the synthesis and release of MT.

摘要

目的

探讨电针(EA)通过褪黑素(MT)抗氧化作用抵抗急性脑缺血再灌注损伤(CI-RI)的效果。

方法

将52只雄性SD大鼠随机分为假手术组(假手术,n = 8)、CI-RI组(模型,n = 8)、EA组(n = 8)、MT组(3.2 mg/Kg,腹腔注射,n = 8)、EA + MT组(n = 8)、EA + 鲁辛朵尔组(L,MT拮抗剂,1 mg/100 μl/大鼠,n = 6)、EA + PD 98059组(丝裂原活化蛋白激酶,MAPK,拮抗剂,1 mg/100 μl/大鼠,n = 6)。采用3 Hz、1 - 3 mA连续波电针“大椎”(GV 14)和“百会”(GV 20)30分钟。除假手术组外,其余组采用右侧大脑中动脉闭塞(MCAO)60分钟再灌注24小时诱导CI-RI模型。采用高效液相色谱法(HPLC)测定松果体组织MT含量,采用Kuluz和Julio法评估大鼠神经行为学评分。采用免疫组织化学法检测纹状体Bax和Bcl-2基因蛋白表达。

结果

与假手术组相比,模型组纹状体Bax免疫反应性(IR)阳性细胞数及Bax/Bcl-2显著增加(P < 0.001),神经行为学评分及Bcl-2 IR阳性细胞数显著降低(P < 0.01)。与模型组相比,EA组松果体MT含量、神经行为学评分及Bcl-2 IR阳性细胞数,EA + L组和EA + P组神经行为学评分,MT组和EA + MT组Bcl-2表达显著增加(P < 0.01,0.001);而EA组、MT组和EA + MT组Bax IR阳性细胞数及Bax/Bcl-2显著降低(P < 0.001)。表明EA可明显增加松果体MT含量,改善CI-RI大鼠活动评分,上调纹状体Bcl-2表达,下调Bax表达。与单纯EA组相比,EA + L组和EA + P组神经行为学评分,EA + MT组Bax IR阳性细胞数及Bax/Bcl-2显著降低(P < 0.05,0.01)。EA + L组和EA + P组行为评分之间,MT组和EA + MT组Bax和Bcl-2 IR阳性细胞数及Bax/Bcl-2之间差异无统计学意义(P > 0.05)。

结论

EA具有良好的抗氧应激作用,可直接或间接保护脑免受缺血损伤,这可能与其上调MT的合成和释放有关。

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