盐酸法舒地尔对脂多糖致大鼠大脑皮质神经元损伤的保护作用及机制研究

摘要

Objective To explore the protective effects of fasudil hydrochloride on injured rat cerebral cortical neurons by lipopolysaccharide ( LPS ) and the possible mechanism. Methods Neurons isolated from newborn rat cerebral cortex were cultured in vitro for seven days , and were divided into five groups randomly : control group, LPS group , and three fasudil hydrochloride of different concentrations ( 50 μmol/L, 100 μmol/L, and 200 μmol/L ) plus LPS groups. The morphologic changes of neurons were observed with inverted phase contrast microscope. The apoptosis and necrosis of neurons were detected by fluorescein stain of Hoechst 33258 , fluorescein stain of AO/EB, and measurement of LDH. The expressions of c - Jun N - terminal kinase ( JNK1 ) were measured by fluorescein stain of fluorescein isothiocyanate. Results Compared with the control group, the neuronal apoptotic ratios were significantly increased in LDP group ( P ＜ 0. 05 ) and the expressions of JNK1 were significantly up - regulated ( P ＜ 0. 05 ). Compared with the LDP group, the neuronal apoptotic ratios were significantly decreased and the expressions of JNK1 were significantly inhibited in three fasudil hydrochloride of different concentrations ( 50 μmol/L, 100 μmol/L, and 200 μmol/L ) plus LPS groups. Conclusion Fasudil hydrochloride has a neuroprotective effect on injured rat cerebral cortical neurons, which may be associated with its effectiveness in resisting apoptosis and inhibiting JNK signaling pathway.%目的 探讨盐酸法舒地尔对脂多糖致大鼠大脑皮质神经元损伤的保护作用及可能机制.方法 提取新生大鼠大脑皮质神经元体外培养,7 d后将培养的神经元随机分为对照组、脂多糖组、不同浓度盐酸法舒地尔(50 μmol/L、100 μmol/L、200 μmol/L)+脂多糖组;采用倒置相差显微镜观察神经元形态学变化,测定乳酸脱氢酶(LDH)活性,并用AO/EB荧光染色、Hoechst 33258荧光染色检测神经元损伤和凋亡情况,FITC荧光染色法测定磷酸化c-Jun氨基端激酶1(JNK1)细胞内表达变化.结果 与对照组相比,脂多糖组可使体外培养神经元凋亡率明显增加(P＜0.05),JNK1表达显著升高(P＜0.05);与脂多糖组比较,不同浓度盐酸法舒地尔+脂多糖组可明显对抗脂多糖引起的神经元凋亡率,JNK1表达上调明显受抑.结论 盐酸法舒地尔对脂多糖诱导大鼠大脑皮质神经元损伤具有保护作用,这可能与抗凋亡、抑制JNK信号传导通路有关.