电针脑缺血大鼠水沟穴调节微小RNA-328促进血管新生的机制研究

摘要

目的 观察电针脑缺血大鼠水沟穴对缺血半暗带区血管新生及微小RNA(miR-328)、CD44mRNA和蛋白表达的影响,探讨电针脑缺血大鼠水沟穴调节miR 328促血管新生的机制.方法 SD雄性大鼠80只,随机分为正常纽、假手术组、模型纽、电针组,每组各5只.Longa法评估各组大鼠神经功能缺损,免疫组织化学染色法观察CD34变化,实时荧光定量PCR技术检测miR-328、CD44 mRNA表达水平,Western blot技术检测CD44蛋白表达水平.结果 假手术组与正常组大鼠在神经功能缺损、CD34为标记的血管新生、miR 328和CD44表达无差异(P＞0.05).与假手术组比较,模型组神经功能缺损、血管新生、miR-328、CD44表达明显增加(P＜＜0.01);与模型组比较,电针刺组神经功能缺损程度改善[(2.6±0.6)分vs (3.4±0.6)分,P＜0.01],血管新生增加显著[(80.40±3.85) vs (67.60±2.79)分,P＜＜0.01],miR-328 1.22±0.37vs 2.02±0.22和CD44 mRNA 4.35±1.33 vs 7.16±1.83,CD44蛋白0.42±0.04 vs 0.55±0.06表达降低(P＜0.05,P＜＜0.01).结论 电针水沟穴可通过调节miR-328及靶基因CD44表达水平,促进缺血半暗带区的血管新生.%Objective To study the effect of electricalal acupuncture of Shuigou (Du 26) on angiogenesis and expression of miR-328,CD44 mRNA and protein in rats with cerebral ischemia.Methods Eighty SD male rats were randomly divided into normol group,sham operation group,model group and electrical acupuncture group (20 in each group).Their neurological function was assessed by Longa occlusion,their CD34 was observed with immunohistochemical staining.Expressions of miR-328,CD44 mRNA and protein were detected by RT-PCR and Western blot respectively.Results The neurological function and angiogenesis scores,the miR-328 and CD44 mRNA and protein expression levels were significantly higher in model group than in sham operation group (P＜0.01).The neurological function scores and angiogenesis were significantly higher while the expression levels of miR-328,CD44 mRNA and protein were significantly lower in electrical acupuncture group than in model group (2.6±0.6 vs 3.4±0.6,P＜0.01;80.40±3.85 vs 67.60±2.79,P＜0.01;1.22±0.37 vs 2.02±0.22,4.35±1.33 vs 7.16±1.83,0.42±0.04 vs 0.55±0.06,P＜0.05,P＜0.01).Conclusion Electricalal acupuncture of Shuigou (Du 26) stimulates angiogenesis in rats with cerebral ischemia by regulating the expression of miR-328 and CD44 mRNA and protein.