大麻素WIN55-212-2对脑出血大鼠皮质内PKAC-β、c-Fos及BDNF表达量的影响(英文)

Changes of PKAC-β,c-Fos and BDNF in cerebral cortex after intracerebral hemorrhage in rats treated with WIN55-212-2

目的:探讨WIN55-212-2对大鼠局灶性脑出血后大脑皮质内PKAC-β、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)和c-Fos mRNA和蛋白质表达的影响。方法:利用VII型胶原酶制作脑出血模型,半小时后腹腔注射WIN55-212-2。实验动物均在术后24h取出脑组织,采用半定量RT-PCR检测大鼠脑皮质内蛋白激酶A(PKAC-β)、c-Fos和BDNF mNRA表达量;采用Western blotting检测PKAC-β和BDNF的蛋白表达量;采用免疫组化方法定位PKAC-β、c-Fos和BDNF蛋白在神经细胞的表达。结果:WIN55-212-2能明显改善脑出血的神经缺失症状,并上调脑出血侧皮质内BDNF和c-Fos的表达,但是抑制PKAC-β的表达。免疫组化结果显示PKAC-β、c-Fos和BDNF蛋白分别在神经元胞膜上、神经元的细胞核上或胶质细胞的胞质中表达。结论:WIN55-212-2不仅可以通过上调转录因子c-Fos mRNA从而增加BDNF的表达量,还可能通过抑制PKA诱导BDNF的表达,发挥神经保护作用。

AIM： To observe the effect of cannabinoid receptor （ CB1R） agonist WIN55-212-2 on the expression of brain-derived neurotrophic factor （ BDNF） ,c-Fos and protein kinase A beta-catalytic subunit （ PKAC-β） in cerebrum cortex after intracerebral hemorrhage （ ICH） in rats. METHODS： The intracerebral hemorrhage model of rat was made by the injection of collagenase Ⅶ,and WIN55-212-2 was intraperitoneally （ ip） injected 30 min later. The rats were killed for sampling the brain tissues as specimens 24 h after ICH. The methods of immunohistochemical analysis and Western blotting were adopted to detect the expression of PKAC-β and BDNF. The mRNA expression of PKAC-β, c-Fos and BDNF was determined by RT-PCR. RESULTS： WIN55-212-2 obviously improved some nervous deficit symptoms and increased the expression of BDNF at mRNA and protein levels with upregulating the mRNA expression of cFos and downregulating the expression of PKA at mRNA and protein levels in the ipsilateral cerebral cortex. The proteins of PKAC-β,c-Fos and BDNF were expressed on the membrane or nucleus of the neuron or in the cytoplasm of glial cells, respectively. CONCLUSION： The expression of BDNF is induced not only by upregulation of c-Fos,but also by downregulation of PKA in WIN55-212-2 treated rats.