摘要
目的研究高糖环境下大鼠肾小球系膜细胞(RMC)的增殖、转化生长因子β1(TGF-β1)的分泌、以及肝细胞生长因子(HGF)的干预作用。方法①将RMC分为3组:正常对照组;高糖组;高糖+HGF组(25.0mmol/L葡萄糖,50ng/mlHGF)。分别培养不同时间(12,24,48,72,96h)。运用MTT法测定细胞的增殖情况。②将RMC分为3组:正常对照组;高糖组;甘露醇对照组:(20.0mmol/L甘露醇)。分别于培养的12、24、48、96h收集细胞及上清夜,用ELISA法来检测TGF-β1分泌量。③将RMC分为:正常对照组;高糖组;高糖+HGF组(HGF浓度分别为25、50、100、200ng/ml)。分别培养48h后,用ELISA法测定此时TGF-β1分泌情况。以上各组正常组中葡萄糖浓度5.5mmol/L,高糖浓度25.0mmol/L。结果①25.0mmol/L高糖在24h促进RMC增殖,肝细胞生长因子(HGF)可以抑制细胞增殖。在48,72和96h高糖抑制细胞增殖。HGF对抗高糖对细胞增殖的抑制作用;而且呈时间依赖性。②高糖促进TGF-β1分泌,HGF可以抑制TGF-β1的分泌,且呈剂量依赖性。结论高糖刺激肾小球系膜细胞TGF-β1出现稳定高表达,而给予外源性的HGF后,系膜细胞TGF-β1的分泌减少,而且呈时间依赖性。
Objective To investigate the effect of high glucose on proliferation and secretion of Transforming growth factor β1（TGF-β1） and in rat mesangial cells（RMC）,and to further study the intervention of Hepatocyte growth factor（HGF）.Methods ①The RMC were allocated into 3 groups randomly：normal glucose group（NG）;high glucose group（HG）;HG ＋ HGF group（25.0 mmol/L glucose＋ 50 ng/ml HGF）.After RMC was cultured for 12,24,48,72,96 h,the proliferation activity of RMC was observed by MTT.②The RMC were allocated into 3 groups：normal contrast group;high glucose group;mannitol contrast group（20.0 mmol/L mannitol）.Collect cells respectively in 12,24,48,96 h of culture,be used to check TGF-β1 secreting amounts.③The RMC were allocated into 3 groups：normal contrast group;high glucose group;HGF group（25.0 mmol/L glucose＋different concentrations of HGF：25.0,50.0,100.0,200.0 ng/ml）.After RMC was cultured with upper three different kinds of environments for 48 hours,the secretion of TGF-β1 was measured by ABC-ELISA.In upper every group normal glucose group（5.5 mmol/L glucose,NG）,high glucose group（25.0 mmol/L glucose,HG）.Results ①25.0 mmol/L glucose could increase the proliferation activity of RMC in 24 h,and HGF could inhibit the proliferation activity of RMC.While high glucose could inhibit the proliferation activity of RMC in 48,72 and 96 h.HGF could increase the proliferation activity of RMC.and shows time-dependently.②The high glucose could promote the secretion of TGF-β1.Conclusion The HGF could promote the proliferation activity of RMC,while HGF could inhibit the secretion of TGF-β1,and shows time-dependently.
出处
《中国现代药物应用》
2011年第17期3-5,共3页
Chinese Journal of Modern Drug Application
