摘要
目的观察叶酸对同型半胱氨酸(homocysteine,Hcy)诱导的人肾脏系膜细胞增殖作用的影响及其可能机制。方法人肾脏系膜细胞采用含5%胎牛血清的RPMI1640培养,采用不同浓度Hcy(200、400、600、800、1 000μmol/L)处理细胞,MTT法观察Hcy对人肾脏系膜细胞增殖的影响。选取理想的Hcy浓度,加入不同浓度叶酸(50、100、200、400μmol/L)处理细胞,观察叶酸对细胞增殖的干预作用。相同方法处理细胞后提取总蛋白,Western blot检测转化生长因子-β(transforming growth factor-β,TGF-β)和细胞外调节蛋白激酶(extracellularregulated protein kinases,ERK)表达。结果 Hcy能诱导人肾脏系膜细胞的增殖,促进TGF-β和ERK蛋白表达,呈现剂量依赖性。叶酸干预后能抑制Hcy诱导的细胞增殖,下调TGF-β和ERK蛋白表达。结论叶酸能抑制Hcy诱导的人肾脏系膜细胞增殖,可能与抑制TGF-β和ERK蛋白表达有关。
Objective This study aimed to investigate the effect of folic acid（FA）on homocysteine（Hcy）- induced proliferation of human renal mesangial cells （HMCs）and possible signaling mechanisms. Methods HMCs were cultured in RPMI1640 medium supplemented with 5 % fetal bovine serum. HMCs were treated with PBS, Hcy （200,400,600,800 and 1 000 υmol/L）or Hcy（400 υmol/L）plus folic acid（50,100,200,400 υmol/L）for 24 hours. Cell viability was assessed by using Methylthiazolyldiphenyl-tetrazolium bromide（MTT）assay and the alterations of transforming growth factor-β（TGF-β）and extracellular regulated protein kinases（ERK）were detected by western blot analysis. Results Hcy significantly stimulated the proliferation of HMCs in a dose dependent manner. However, Hcy-induced proliferation of HMCs was attenuated by additional supplement of FA. Protein levels of TGF- β and ERK were increased in the presence of Hcy,which was reversed by addition of FA. Conclusion FA attenuated Hcy-induced proliferation of HMCs,which was associated with decreased expression of TGF-β and ERK orotein.
出处
《成都医学院学报》
CAS
2013年第2期149-152,共4页
Journal of Chengdu Medical College
基金
国家自然科学基金项目(NO:30600250)
广东省自然科学基金项目(NO:10151008901000146)
高校基本科研业务费中山大学青年教师培育项目(NO:10ykpy33)
关键词
同型半胱氨酸
叶酸
转化生长因子-Β
细胞外调节蛋白激酶
人肾脏系膜细胞
Homocysteine
Folic acid
Transforming growth factor-!3
Extracellular regulated protein kinasesHuman renal mesangial cells
