针刺对脑卒中大鼠炎症损伤及miR-214的影响

目的　探究针刺对脑卒中大鼠炎症损伤及微小RNA（miR）-214的影响。方法　改良Longa线栓法建立脑卒中大鼠模型36只，建模成功后随机数字表法分为模型组、针刺组（针刺阳陵泉、曲池穴）、阳性对照组（0.4 mg/kg尼莫地平），另设假手术组，每组12只。Longa评分评价大鼠行为学情况；2，3，5-氯化三苯基四氮唑（TTC）染色检测大鼠脑梗死体积；苏木精-伊红（HE）染色观察大鼠大脑冠状部情况；实时荧光定量PCR（qPCR）检测脑组织miR-214水平；蛋白免疫印迹检测大鼠大脑冠状部第10号染色体上磷酸酶和张力蛋白同源缺失的基因（PTEN）、丝苏氨酸蛋白激酶（AKT）、磷酸化AKT（p-AKT）、核因子-κB（NF-κB）、白细胞介素-6（IL-6）蛋白水平。结果　与假手术组比较，模型组炎症细胞浸润、纤维化现象严重，Longa评分、脑梗死体积、miR-214表达水平、p-AKT/AKT以及IL-6、细胞核NF-κB蛋白表达水平升高（P＜0.05）；PTEN、细胞浆NF-κB蛋白表达水平降低（P＜0.05）。与模型组比较，针刺组和阳性对照组炎症和纤维化现象减轻，Longa评分、miR-214表达水平、p-AKT/AKT、细胞核NF-κB以及IL-6蛋白表达水平降低，且针刺组脑梗死体积减小（P＜0.05）；PTEN、细胞浆NF-κB蛋白表达水平升高（P＜0.05）。结论　针刺脑卒中大鼠可通过抑制miR-214及下游信号通路来减轻大脑冠状部炎症损伤。

The effect of acupuncture on inflammatory damage and miR-214 in stroke model rats

Objective To explore the effects of acupuncture on inflammatory damage and microRNA (miR)-214 in stroke model rats. Methods Modified Longa thread suppository method was used to establish 36 rat models of stroke. After successful modeling, rats were randomly divided into model group, acupuncture group (acupuncture at Yanglingquan and Quchi points) and positive control group (0.4 mg/kg nimodipine). Twelve rats were used as the sham operation group. Longa score was used to evaluate the behavior of rats. The volume of cerebral infarction was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin eosin (HE) staining was used to observe the coronary region of the brain of rats. The level of miR-214 in brain tissue was detected by real-time fluorescence quantitative PCR (qPCR). Western blot assay was used to detect the phosphatase and tensin homologous deletion gene on chromosome ten (PTEN), threonine protein kinase (AKT), phosphorylated AKT (p-AKT), nuclear factor κB (NF-κB) and interleukin-6 (IL-6) proteins in the coronary region of rat brain. Results Compared with the sham operation group, the inflammatory cell infiltration and fibrosis were severe in the model group. Longa score, cerebral infarction volume, miR-214 level, p-AKT/AKT, IL-6 and nuclear NF-κB protein levels increased (P＜0.05). PTEN and cytoplasmic NF-κB protein levels decreased (P＜0.05). Compared with the model group, the inflammation and fibrosis were reduced in the acupuncture group and positive control group. Longa score, miR-214 level, p-AKT/AKT, nuclear NF-κB and IL-6 protein levels decreased, and cerebral infarction volume decreased in the acupuncture group (P＜0.05). PTEN and cytoplasmic NF-κB protein levels increased (P＜0.05). Conclusion The acupuncture can reduce the inflammatory damage of cerebral coronary brain tissue by inhibiting miR-214 and downstream signal pathways in stroke model rats.