摘要
目的研究山茱萸活性成分马钱苷、莫诺苷对D-半乳糖致衰大鼠星形胶质细胞的保护作用。方法常规方法体外培养乳大鼠大脑皮层星形胶质细胞,采用免疫荧光方法进行星形胶质细胞鉴定;MTT法对比不同浓度D-半乳糖对星形胶质细胞的增殖抑制作用,以及马钱苷、莫诺苷对D-半乳糖致衰星形胶质细胞活力的影响;以超氧化物歧化酶(SOD)、丙二醛(MDA)为观察指标,观察马钱苷、莫诺苷对D-半乳糖致衰星形胶质细胞的影响;ELISA法检测胶质细胞源性神经营养因子(GDNF)、碱性成纤维细胞生长因子(bFGF)含量变化;Western blot法检测Bax、caspase-3、磷酸化细胞外信号调节激酶(p-ERK1/2)、磷酸化丝裂原活化蛋白激酶/ERK激酶(p-MEK1/2)的表达水平,以做进一步机制研究。结果马钱苷、莫诺苷组与模型组比较,细胞的增殖能力提高,SOD含量增加,MDA活性降低,且差异有显著性(P<0.05);马钱苷、莫诺苷组生长因子(GDNF、bFGF)的分泌高于模型组;Western blot结果表明,D-半乳糖致衰老的星形胶质细胞中Bax、caspase-3蛋白表达无明显变化,模型组pERK1/2、p-MEK1/2蛋白表达降低,马钱苷、莫诺苷增加pERK1/2、p-MEK1/2蛋白的表达。结论山茱萸活性成分马钱苷、莫诺苷可通过提高D-半乳糖致衰星形胶质细胞的增殖能力,提高衰老细胞的抗氧化能力,发挥抗衰老作用,其机制可能与MEK/ERK介导的信号通路调控有关。
Aim To study effects of loganin and morro- niside on aging astrocytes induced by D-galactose. Methods Cortex astrocytes of newly born rats were cultured in vivo and indentified by immunofluorescence method. Firstly, appropriate D-galactose concentrationwas selected and effects of loganin and morroniside on proliferation activity of aging astrocytes induced by D- galactose were determined by MTT test. Then SOD, MDA were taken as indicators to study the effects of loganin and morroniside on aging astrocytes inducedby D-galactose. Thirdly, growth factors like gliar cell line-derived neurotrophic factor （GDNF）, basic fibro- blast growth factor （bFGF） tested by ELISA method and expression of Bax, caspase-3, phospho-extracellu- lar signal-regulated kinese （p-ERK1/2） , phospho-mi- togen-actived protein kinase/ extracellular signal-regu- lated kinase kinase （p-MEK1/2） were taken as indi- cators to discuss the potential protection mechanism. Results Loganin and morroniside exerted certain effects on proliferation of aging astrocytes induced by D-galactose, improving SOD, GDNF, bFGF release and lowering MDA release significantly （ P 〈 0. 05 ）. The expressions of Bax and caspase-3 proteins had nodifference between model group and loganin group, morroniside group. While the expressions of p-ERK1/ 2, p-MEK1/2 of loganin group, morroniside group were improved significantly compared with model group. Conclusion Loganin and morroniside have protective effects on aging astrocytes induced by D-ga- lactose, and increase the proliferation ability. Protec- ting their antioxidant systems and improving the expres- sion of p-ERK1/2, p-MEK1/2 proteins are possible mechanisms.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2014年第12期1688-1692,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 30772851)
江苏省“六大人才高峰”第六批资助项目
江苏高校优势学科建设工程资助项目(PAPD)
