CAJ | 학술논문

目的探讨甲状旁腺激素(PTH)对人近曲小管上皮细胞系HK-2细胞分泌结缔组织生长因子(CTGF)及细胞转分化的影响.方法应用实时定量PCR和Western印迹观察PTH诱导HK-2细胞CTGF mRNA和蛋白表达情况,从转录水平探讨PTH对CTGF基因启动子活性的调控作用.应用免疫荧光技术检测PTH作用下HK-2细胞中α平滑肌肌动蛋白(α-SMA)的表达.结果 HK-2细胞有基础水平的CTGF mRNA和蛋白表达,PTH刺激后其表达量显著增加.FTH最佳刺激浓度是10-10 mol/L,最佳刺激时间是72 h.10-10 mol/L PTH干预HK-2细胞12 h后,荧光素酶活性较对照组显著升高(1 .888±0 .078比0 .989±0 .030,P＜O .01);光镜下可见细胞由立方形铺路石样转变为梭形,随作用时间延长转分化现象更显著.PIM刺激HK-2细胞12 h后,免疫荧光检测可见细胞质中有α-SMA表达;PTH刺激24 h后,α-SMA表达明显增多,与CTGF mRNA和蛋白表达的时间效应一致.结论 PTH可上调HK-2细胞CTGF表达,并具有促进HK-2细胞转分化的作用.
목적 탐토갑상방선격소(PTH)대인근곡소관상피세포계HK-2세포분비결체조직생장인자(CTGF)급세포전분화적영향.방법 응용실시정량PCR화Western인적관찰PTH유도HK-2세포CTGF mRNA화단백표체정황,종전록수평탐토PTH대CTGF기인계동자활성적조공작용.응용면역형광기술검측PTH작용하HK-2세포중α평활기기동단백(α-SMA)적표체.결과 HK-2세포유기출수평적CTGF mRNA화단백표체,PTH자격후기표체량현저증가.FTH최가자격농도시10-10 mol/L,최가자격시간시72 h.10-10 mol/L PTH간예HK-2세포12 h후,형광소매활성교대조조현저승고(1 .888±0 .078비0 .989±0 .030,P＜O .01);광경하가견세포유립방형포로석양전변위사형,수작용시간연장전분화현상경현저.PIM자격HK-2세포12 h후,면역형광검측가견세포질중유α-SMA표체;PTH자격24 h후,α-SMA표체명현증다,여CTGF mRNA화단백표체적시간효응일치.결론 PTH가상조HK-2세포CTGF표체,병구유촉진HK-2세포전분화적작용.
Objective To investigate the effect of parathyroid hormone (PTH) on the transition and connective tissue growth factor (CTGF) expression of human renal proximal tubular epithelial cell line HK-2 . Methods The expression of CTGF mRNA and protein of HK-2 cells were measured by real time RT-PCR and Western blot respectively . The effect of PTH on the phenotypic transformation of HK-2 cells was examined by light microscopy . The expression of α-smooth muscle actin (α-SMA) in HK-2 cells was detected by immunofluorescence . Results Basal level of CTGF mRNA and the protein expression were detected in HK-2 ceils . PTH upregulated the expression of CTGF mRNA and protein with the maximal response at the concentration of 10-10 mol/L and the best stimulating time was at 72 h . After exposure to PTH (10-10tool/L) for 12 hours, the highest level of luciferase activity was 1 .96 fold as compared to control (1 .888±0 .078 vs 0 .989±0 .030, P＜0 .01 ) . Untreated cells showed negligible expression of ±-SMA,whereas ±-SMA expression was significantly increased in cells treated with PTH . Conclusion PTH up-regulates CTGF expression and induces transition of HK-2 cells .