大麻素对脑出血大鼠皮质内PKAC-β、c-fos及BDNF表达量的影响

Effects of cannabinoid on the expression of PKAC-β,c-fos and BDNF in cerebral cortex after ICH

目的探讨大麻素(WIN55,212-2)对大鼠局灶性脑出血后同侧大脑皮质内PKAC-β、c-fos和BDNF mRNA及蛋白质表达量的影响。方法采用Ⅶ型胶原酶制作大鼠脑出血模型,术后0.5h后腹腔注射药物,均在24h后取脑组织;采用免疫组化方法定位PKAC-β、c-fos和BDNF蛋白;采用RT-PCR检测各组大脑皮质内PKAC-β、c-fos和BDNF mRNA的表达;采用Western blot检测各组大脑皮质内PKAC-β和BD-NF蛋白质的表达。结果WIN55,212-2上调同侧大脑皮质内BDNF mRNA和蛋白质的表达,同时也上调c-fos mRNA表达量,但抑制PKAC-β mRNA和蛋白质的表达;免疫组化结果显示PKAC-β,c-fos和BDNF蛋白分别位于神经元胞膜、细胞核或胶质细胞的胞质中。结论WIN55,212-2促进大脑皮质内BDNF蛋白的合成,为其临床上治疗脑血管病提供理论依据。

Aim To explore the effects of cannabinoid （WIN55,212-2） on mRNA and protein expressions of PKAC-β,c-fos and BDNF in cerebral cortex after intracerebral hemorrhage（ICH）in rats.Methods ICH model of rats were made by Ⅶ Collagenase,which were injected by the intraperitoneal route,thirty minutes after the operations.The rats were killed for brain tissue as specimens with 24 hours.The localizations of PKAC-β,c-fos and BDNF were assessed by immunohistochemistry;The expressions of PKAC-β,c-fos and BDNF mRNA were detected by RT-PCR;The expressions of PKAC-β and BDNF protein were revealed by Western blot.Results WIN55,212-2 increased not only the levels of BDNF mRNA and protein,but also c-fos mRNA in ipsilateral cerebral cortex.However,it decreased the levels of PKAC-β mRNA and protein.PKAC-β,c-fos,and BDNF proteins were expressed on membrane of neurons,nucleus of neurons or the cytoplasm of glial cells respectively.Conclusion WIN55,212-2 induces the expression of BDNF in the cerebral cortex,which provides a theoretical basis for the treatment of cerebrovascular disease.