CAJ | 학술논문

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mRNA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mRNA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。
배경：혈소판연화생장인자가유도신소구계막세포급신간질다충세포증식、천이、전화급세포외기질적과표체。목적：관찰혈소판연화생장인자DD/혈소판연화생장인자βR신호통로격활대대서신성섬유세포세포증식급α-평활기기동단백표체적영향。방법：체외배양정상대서신성섬유세포(NRK-49F)，안조혈소판연화생장인자DD자격질량농도분위대조조、1μg/L조、10μg/L조、50μg/L조、100μg/L조，연후근거50μg/L 혈소판연화생장인자DD자격불동시간분위대조조、12 h조、24 h조、48 h조。CCK8검측불동혈소판연화생장인자DD질량농도급혈소판연화생장인자DD작용불동시간대NRK-49F증식적영향；Real-time PCR검측각질량농도조혈소판연화생장인자βR、α-평활기기동단백mRNA변화；Western blot검측각농도조혈소판연화생장인자βR、α-평활기기동단백단백표체변화。결과여결론：여대조조상비，혈소판연화생장인자DD가명현자격NRK-49F세포증식，정현제량의뢰성급시간의뢰성。혈소판연화생장인자DD가제량의뢰성자격기상응혈소판연화생장인자βR급α-평활기기동단백재mRNA수평급단백수평상적표체증가。제시혈소판연화생장인자DD/혈소판연화생장인자βR신호통로적격활가현저자격NRK-49F세포적증식급향기성섬유세포적전화。
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 <br> 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.