李氏5号方对D-半乳糖老化小鼠海马神经元NGF信号转导的影响

通过检测D 半乳糖老化小鼠海马神经元NGF(神经生长因子 )、PI3K(磷脂酰肌醇酯 3激酶 )、Akt/PKB(蛋白激酶B)、CREB(cAMP应答元件结合蛋白 )的表达 ,观察李氏 5号方对海马神经元NGF信号转导的影响。将小鼠随机分为 5组 :正常对照组 (C组 )、D 半乳糖模型组 (D组 )、李氏 5号方大剂量药物治疗组 (L组 )、中剂量药物治疗组(M组 )、小剂量药物治疗组 (S组 )。D、L、M、S 4组每日皮下注射半乳糖 65mg/kg。L、M、S 3个治疗组分别灌胃李氏5号方 0 .6、0 .3、0 .1 5g/kg ,连续 3个月。免疫组织化学染色方法和Westernblotting检测海马神经元NGF、PI3K、Akt/PKB、CREB的表达。结果 :4种抗体C组深染的阳性细胞在海马广泛分布 ,细胞计数与D组有显著性差异 ;D组海马阳性反应细胞着色浅 ,少有突起 ,细胞数目少。 3个剂量李氏 5号方治疗组染色结果与C组相似 ,与半乳糖老化组比较均有显著性差异 ,李氏 5号方水提取液能明显上调NGF的PI3K通路中的一些信号转导蛋白 ,提示 :李氏 5号方水提取液中可能含有能激活PI3K的活性物质 ,为深入研究李氏 5号方的有效成分打下基础。

Effect of Lishi No.5 Formula Liquid Extract on NGF Signal Transduction Pathway of D-galactose Induced Aging of Murine Brain

The aim was to observe the the effect of Lishi No.5 Formula liquid extract on NGF signal transduction pathway of D-galactose induced aging of murine brain(DGAM). Kunming mice were divided into 5 groups: control group(C), DGAM group(D), Lishi No.5 Formula large dose treatment group(L), medium dose treatment group(M), and small dose treatment group(S). Immunohistochemical staining and Western Blotting for NGF, PI3K, Akt/PKB, CREB expression of hippocampal neurons were done. The expression level of NGF, PI3K, Akt/PKB, CREB in the hippocampal decreased obviously in DGAM group compared with control group, while the result of Chinese medicine groups were the same as that in normal mice. Chinese medicine Lishi No.5 Formula up-regulate PI3K signal transduction pathway of NGF in the hippocampal of DGAM group, which showed Lishi No.5 Formula liquid might contain active substance which could stimulate PI3K. This research forms basis for studying the utility elements of Lishi No.5 Formula.