摘要
背景:血小板衍化生长因子可诱导肾小球系膜细胞及肾间质多种细胞增殖、迁移、转化及细胞外基质的过表达。目的:观察血小板衍化生长因子DD/血小板衍化生长因子βR信号通路激活对大鼠肾成纤维细胞细胞增殖及α-平滑肌肌动蛋白表达的影响。方法:体外培养正常大鼠肾成纤维细胞(NRK-49F),按照血小板衍化生长因子DD刺激质量浓度分为对照组、1μg/L组、10μg/L组、50μg/L组、100μg/L组,然后根据50μg/L血小板衍化生长因子DD刺激不同时间分为对照组、12 h组、24 h组、48 h组。CCK8检测不同血小板衍化生长因子DD质量浓度及血小板衍化生长因子DD作用不同时间对NRK-49F增殖的影响;Real-time PCR检测各质量浓度组血小板衍化生长因子βR、α-平滑肌肌动蛋白m RNA变化;Western blot检测各浓度组血小板衍化生长因子βR、α-平滑肌肌动蛋白蛋白表达变化。结果与结论:与对照组相比,血小板衍化生长因子DD可明显刺激NRK-49F细胞增殖,呈现剂量依赖性及时间依赖性。血小板衍化生长因子DD可剂量依赖性刺激其相应血小板衍化生长因子βR及α-平滑肌肌动蛋白在m RNA水平及蛋白水平上的表达增加。提示血小板衍化生长因子DD/血小板衍化生长因子βR信号通路的激活可显著刺激NRK-49F细胞的增殖及向肌成纤维细胞的转化。
BACKGROUND：Platelet-derived growth factors can induce proliferation, migration, transformation and extracelular matrix expression of glomerular mesangial cels and renal interstitial cels. OBJECTIVE：To investigate the effect of activation of platelet-derived growth factor-DD/βR signaling pathway on the proliferation and α-smooth muscle actin expression in rat renal fibroblasts. METHODS： Normal rat kidney interstitial fibroblasts （NRK-49F） cultured in vitro were divided into the folowing groups by platelet-derived growth factor-DD concentrations： control, 1 μg/L, 10 μg/L, 50 μg/L, 100 μg/L. And NRK-49Fs were then divided into four groups according to the stimulation time of 50 μg/L platelet-derived growth factor-DD：control, 12 hours, 24 hours, 48 hours. The cel viability of the NRK-49Fs was assessed by Cel Counting Kit-8 after platelet-derived growth factor-DD administration. The mRNA and protein expression levels of
platelet-derived growth factor-βR and α-smooth muscle actin in al groups stimulated by different concentrations of platelet-derived growth factor-DD were determined by RT-PCR and western blot, respectively. RESULTS AND CONCLUSION：Platelet-derived growth factor-DD could facilitate the proliferation rates of the NRK-49Fs at a dose- and time-dependent manner as compared with the control groups. Platelet-derived growth factor-DD could stimulate the mRNA and protein expressions of platelet-derived growth factor-βR and α-smooth muscle actin in a dose-dependent manner. This suggest that the activation of platelet-derived growth facto-DD/βR signaling pathway can obviously promote the proliferation of NRK-49Fs as wel as tranformation into myofibroblasts.
出处
《中国组织工程研究》
CAS
北大核心
2015年第11期1688-1693,共6页
Chinese Journal of Tissue Engineering Research
基金
青岛市科技计划基础研究项目[11-2-4-2-(12)-jch]~~
