白藜芦醇通过介导eNOS表达促进局灶脑缺血/再灌注大鼠脑内血管再生  

Resveratrol promotes angiogenesis via eNOS in the brains of rats with focal cerebral ischemia and reperfusion

作　　者：李杏芮 陈清[1] 盛华均[1] Li Xingrui Chen Qing Sheng Huajun(Anatomy Teaching and Research Section of Chongqing Medical University, Chongqing 400016, China)

机构地区：[1]重庆医科大学解剖学教研室,重庆400016

出　　处：《中国组织化学与细胞化学杂志》2016年第6期506-511,522,共7页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：目的探讨eNOS在白藜芦醇促进局灶脑缺血/再灌注大鼠大脑缺血皮质区血管再生中的作用。方法 80只SD雄性大鼠随机分为假手术组(Sham组)、模型组(I/R组)、模型+白藜芦醇组(I/RB组)、模型+白藜芦醇+eNOS特异性拮抗剂L-NAME组(I/RBL组)。采用线栓法制备大鼠局灶脑缺血/再灌注模型,再灌注后2h后腹腔注射白藜芦醇,连续7d,以再灌注后24h、48h、7d为观察时相点。对I/R、I/RB和I/RBL组大鼠行改良神经功能缺损程度评分,HE染色观察大脑缺血皮质区病理结构变化,Western Blot检测eNOS蛋白表达,免疫组织化学检测VEGF、CD34表达情况,荧光定量PCR检测eNOS mRNA表达。结果 I/RB组再灌注后各时间点大鼠大脑缺血皮质区eNOS蛋白及mRNA、VEGF蛋白表达较I/R组明显升高,侧脑室注射L-NAME阻断eNOS作用后,I/RBL组eNOS、VEGF表达较I/RB组降低。同时,白藜芦醇可有效促进缺血后神经功能恢复、改善缺血损伤后脑组织病理变化,增加CD34^+微血管密度。结论白藜芦醇可能通过上调局灶脑缺血/再灌注大鼠大脑缺血皮质区eNOS和VEGF表达,促进脑微血管再生,发挥缺血损伤后脑保护作用。Objective To investigate the role of eNOS in the promotion of angiogenesis by resveratrol in the iscbemic cerebral cortex of rats after focal cerebral ischemia and reperfusion. Methods 80 healthy male adult Sprague Dawley （SD） rats were randomly divided into 4 groups： the sham group （Sham）, model group （I/R）, resveratrol group （I/RB） and I/RB plus L-NAME （ a specific antagonist of eNOS） group （ I/RBL）. The SD rat models of focal cerebral ischemia and reperfusion were prepared by the thread embolism method. Resveratrol was given by intra-peritoneal injection once a day for 7 days, the first at 2h after reperfusion. Tests were performed at 24h, 48h and 7d after reperfusion. The score of modified neurological severity was assessed in I/R, I/RB and I/RBL groups, and the pathological change in the structure of the ischemic cerebral cortex was observed by HE staining. The eNOS protein and mRNA were detected by Western Blot and FQ-PCR, respectively, and the expression of VEGF and CD34 was examined by immunohistochemistry. Results The expression of eNOS protein and mRNA and VEGF protein in the ischemic cerebral cortex of rats was significantly higher in I/RB than in I/R at all the three time points, and that in I/RBL with lateral ventricle injection of L-NAME to block the effect of eNOS was obviously lower than in I/RB. Moreover, resveratrol treatment after occlusion facilitated the recovery of neurological functions, improved the structure of ischemic brain tissue and increased the density of CD34^＋ microvessels. Conclusion Resveratrol can promote angiogenesis in the brains of rats with focal cerebral ischemia and reperfusion to protect isehemic brain tissue by up-regulating the expression of eNOS and VEGF in the ischemic cerebral cortex.

关 键 词：局灶脑缺血/再灌注 白藜芦醇 L-NAME ENOS VEGF 脑保护 

分 类 号：R743[医药卫生—神经病学与精神病学]