眼针疗法对急性脑缺血再灌注损伤大鼠脑组织水通道蛋白4表达的影响

[Effects of eye-acupuncture therapy on the expression of AQP4 in brain tissue of rats with acute cerebral ischemia-reperfusion injury].

作者信息

Wang Zhe, Ma Xian-De, Liu Hui-Hui, Liu Xu-Dong, Gao Yuan, Guan Hong-Quan, Wang De-Shan

机构信息

Liaoning University of TCM, Shenyang 110032, China.

出版信息

Zhongguo Zhen Jiu. 2011 Aug;31(8):727-31.

PMID:21894701

Abstract

OBJECTIVE

To explore the mechanism of the eye-acupuncture for treatment of acute cerebral ischemia-reperfusion injury.

METHODS

Thirty-two healthy SD rats were randomly divided into a normal group, a sham operation group, a model group and an eye-acupuncture group, 8 rats in each group. The rat model of cerebral ischemia-reperfusion was established with thread occlusion method in the model group and the eye-acupuncture group. The eye-acupuncture group was treated by eye-acupuncture at "liver region", "upper energizer area", "lower energizer area" and "kidney region" for 20 min immediately after reperfusion and at 30 min before sampling. No treatment was done in the normal group and the sham operation group, and no thread occlusion was performed in the sham operation group. The Neurologic impairment was scored and the methods of immunohistochemistry staining, western-blotting and real-time fluorescent quantitation polymerase chain reaction (RQ-PCR) were taken to detect the expression of the aquaporin protein 4 (AQP4) and its mRNA in cerebral cortex after reperfusion for 3 hours.

RESULTS

The neurologic impairment score of 1.50 +/- 0.54 in the eye-acupuncture group was significant lower than 2.63 +/- 0.92 in the model group (P < 0.01). The expression of the AQP4 protein by immunohistochemistry and western-blot respectively were 116.33 +/- 10.24 and 0.53 +/- 0.04 in the normal group, 118.97 +/- 12.72 and 0.55 +/- 0.07 in the sham operation group, and 129.30 +/- 18.36 and 0.67 +/- 0.08 in the eye-acupuncture group, with statistical significance compared to 150.88 +/- 15.82 and 0.94 +/- 0.04 in the model group (all P < 0.01), and there were significant differences between the eye-acupuncture group and the normal group (both P < 0.01). The tendency in the expression of AQP4 protein and its mRNA in all the group were almost the same.

CONCLUSION

The eye-acupuncture therapy can relieve the cerebral ischemia-reperfusion injury and the protective mechanism is related to the downregulation of the cerebral AQP4 expression.

摘要

目的

探讨眼针治疗急性脑缺血再灌注损伤的机制。

方法

将32只健康SD大鼠随机分为正常组、假手术组、模型组和眼针组，每组8只。模型组和眼针组采用线栓法制备脑缺血再灌注大鼠模型。眼针组于再灌注后立即针刺“肝区”“上焦区”“下焦区”及“肾区”20分钟，并于取材前30分钟针刺一次。正常组和假手术组不做处理，假手术组不进行线栓。对神经功能缺损进行评分，采用免疫组织化学染色、蛋白质免疫印迹法及实时荧光定量聚合酶链反应（RQ-PCR）检测再灌注3小时后大脑皮质水通道蛋白4（AQP4）蛋白及其mRNA的表达。

结果

眼针组神经功能缺损评分为1.50±0.54，明显低于模型组的2.63±0.92（P<0.01）。免疫组织化学法及蛋白质免疫印迹法检测正常组AQP4蛋白表达分别为116.33±10.24、0.53±0.04，假手术组分别为118.97±12.72、0.55±0.07，眼针组分别为129.30±18.36、0.67±0.08，与模型组的150.88±15.82、0.94±0.04相比差异均有统计学意义（均P<0.01），且眼针组与正常组相比差异均有统计学意义（均P<0.01）。各组AQP4蛋白及其mRNA表达趋势基本一致。