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针刺百会(GV 20)和足三里(ST 36)对脑缺血再灌注损伤大鼠外周血清中微小RNA 124、层粘连蛋白和整合素β1表达的影响。

Effects of acupuncture at Baihui (GV 20) and Zusanli (ST 36) on peripheral serum expression of MicroRNA 124, laminin and integrin β1 in rats with cerebral ischemia reperfusion injury.

作者信息

Chen Su-Hui, Sun Hua, Zhang Ya-Min, Xu Hong, Yang Yang, Wang Fu-Ming

机构信息

Department of Traditional Chinese Medicine, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100730, China.

出版信息

Chin J Integr Med. 2016 Jan;22(1):49-55. doi: 10.1007/s11655-015-2112-7. Epub 2015 Oct 1.

Abstract

OBJECTIVE

To explore the effects of acupuncture at Baihui (GV 20) and Zusanli (ST 36) on the peripheral serum expression of microRNA 124 (miRNA 124), laminin and integrin β1 in rats with cerebral ischemia reperfusion injury (CIRI).

METHODS

Seventy-two healthy male Sprague-Dawley rats were randomized into a model group, an acupuncture group, and a sham-operated group using a random digits table, with 24 rats per group. Each group was further randomly divided into 1-, 3-, 5-, and 7-day subgroups based on the reperfusion time according to a random digits table, with 6 rats in each subgroup. In the model and acupuncture groups, CIRI was induced using the thread occlusion method. Electroacupuncture stimulation was applied daily to GV 20 and left ST 36 for 20 min at the indicated time points after successful operations. Serum was sampled for detecting laminin and integrin β1 protein via enzyme-linked immunosorbent assay, and serum miRNA 124 was examined using quantitative polymerase chain reaction.

RESULTS

The serum level of miRNA 124 in the cerebral ischemia rats increased significantly, and the peak expression of miRNA 124 in both the model and acupuncture groups occurred at 3 days. The expression of miRNA 124 in the acupuncture group was higher than in the model group at the same time point (5.96±0.01 vs. 3.11±0.04, P <0.05). Laminin expression in serum from the cerebral ischemia group was higher than that in the sham-operated group. Compared with the model group, the level of laminin in the serum of the acupuncture group was significantly lower at each time point, especially at the 3-day, and 7-day time points (589.12±3.57 vs. 793.05±5.28, and 600.53±3.05 vs. 899.06±5.74, P <0.05). The level of integrin β1 in the serum from the acupuncture group was lower than that in the model group particularly at the 3-day and 7-day time points (208.66±0.95 vs. 280.83±1.77, and 212.36±0.95 vs. 316.77±2.42, P <0.05). Additionally, the model group and the acupuncture group showed dual peaks of integrin β1 and laminin expression at 3-day and 7-day.

CONCLUSIONS

Acupuncture at GV 20 and ST 36 in rats alleviated CIRI and was associated with upregulated expression of miRNA 124 and with downregulated expression of integrin β1 and laminin in peripheral serum. These changes may represent one of the mechanisms underlying acupuncture's attenuation of CIRI.

摘要

目的

探讨针刺百会(GV 20)和足三里(ST 36)对脑缺血再灌注损伤(CIRI)大鼠外周血清中微小RNA 124(miRNA 124)、层粘连蛋白和整合素β1表达的影响。

方法

将72只健康雄性Sprague-Dawley大鼠采用随机数字表法随机分为模型组、针刺组和假手术组,每组24只。再根据再灌注时间,每组按随机数字表法进一步随机分为1天、3天、5天和7天亚组,每个亚组6只。模型组和针刺组采用线栓法制备CIRI模型。造模成功后,在各时间点对GV 20和左侧ST 36每日进行电针刺激20分钟。采集血清,采用酶联免疫吸附测定法检测层粘连蛋白和整合素β1蛋白,采用定量聚合酶链反应检测血清miRNA 124。

结果

脑缺血大鼠血清miRNA 124水平显著升高,模型组和针刺组miRNA 124的表达峰值均出现在3天。针刺组在同一时间点miRNA 124的表达高于模型组(5.96±0.01 vs. 3.11±0.04,P <0.05)。脑缺血组血清中层粘连蛋白的表达高于假手术组。与模型组相比,针刺组血清中层粘连蛋白水平在各时间点均显著降低,尤其是在3天和7天时间点(589.12±3.57 vs. 793.05±5.28,600.53±3.05 vs. 899.06±5.74,P <0.05)。针刺组血清中整合素β1水平低于模型组,尤其在3天和7天时间点(208.66±0.95 vs. 280.83±1.77,212.36±0.95 vs. 316.77±2.42,P <0.05)。此外,模型组和针刺组整合素β1和层粘连蛋白表达在3天和7天出现双峰。

结论

针刺大鼠GV 20和ST 36可减轻CIRI,且与外周血清中miRNA 124表达上调、整合素β1和层粘连蛋白表达下调有关。这些变化可能是针刺减轻CIRI的机制之一。

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