CAJ | 학술논문

目的观察甲状旁腺激素(PTH)对人肾小管上皮细胞分泌结缔组织生长因子(CTGF)的影响,并探讨丝裂原活化蛋白激酶(MAPK)信号途径在此过程中的作用.方法采用实时定量PCR、Western印迹、报告基因等技术,观察PTH诱导人近端肾小管上皮细胞系HK-2细胞CTGF表达的情况.使用信号通路抑制剂PD98059、U0126阻断信号通路以明确PIH发挥作用的信号途径.结果正常HK-2细胞有基础水平的CTGF mRNA和蛋白表达,PIH刺激后其表达水平显著增加.10-10mol/L PTH作用12 h后,荧光素酶活性较对照组明显升高[(1.8884±0.0780)比(0.9891±0.0300)A,P<0.01].正常HK-2细胞有少量p-ERK1/2表达,PTH刺激后p-ERK1/2表达明显升高,以10-10mol/L PTH作用30 min时效应最强;MAPK通路抑制剂PD98059、U0126作用后,CTGF mRNA、蛋白、基因启动子表达均明显下降.结论 PrH可诱导HK-2细胞CTGF表达,其作用可能是通过MAPK信号通路来实现的.
목적 관찰갑상방선격소(PTH)대인신소관상피세포분비결체조직생장인자(CTGF)적영향,병탐토사렬원활화단백격매(MAPK)신호도경재차과정중적작용.방법 채용실시정량PCR、Western인적、보고기인등기술,관찰PTH유도인근단신소관상피세포계HK-2세포CTGF표체적정황.사용신호통로억제제PD98059、U0126조단신호통로이명학PIH발휘작용적신호도경.결과 정상HK-2세포유기출수평적CTGF mRNA화단백표체,PIH자격후기표체수평현저증가.10-10mol/L PTH작용12 h후,형광소매활성교대조조명현승고[(1.8884±0.0780)비(0.9891±0.0300)A,P<0.01].정상HK-2세포유소량p-ERK1/2표체,PTH자격후p-ERK1/2표체명현승고,이10-10mol/L PTH작용30 min시효응최강;MAPK통로억제제PD98059、U0126작용후,CTGF mRNA、단백、기인계동자표체균명현하강.결론 PrH가유도HK-2세포CTGF표체,기작용가능시통과MAPK신호통로래실현적.
Objective To evaluate the effect of parathyroid hormone (PTH) on the expression of connective tissue growth factor (CTGF) in human renal tubular epithelial cells, and to explore the role of MAPK signaling pathway. Methods Real time RT-PCR, Western blot, and reporter gene assay were employed to detect PTH-induced CTGF expression in HK-2 cells. Inhibitors (PD98059 and U0126) of MAPK signaling pathway were used to confirm involved signal pathway. Results HK-2 cells had basic expression level of CTGF mRNA and protein, which were increased significantly after treatment with PTH. The luciferase activity was up-regulated to a higher level as compared with control group after treatment with 10-10 mol/L PTH for 12 h [(1.8884±0.0780) vs (0.9891±0.0300) A, P<0.01]. Moreover, a small amount of p-ERK1/2 was detected in normal HK-2 cells, but it was increased significantly in response to PTH activation, most remarkably when treated with 10-10 mol/L PTH for 30 min. Inhibitors of MAPK signaling pathway, PD98059 and U0126, noticeably inhibited the expression of CTGF mRNA and protein as well as gene promoters in HK-2 cells. Conclusion PTH can induce higher expression of CTGF in HK-2 cells probably via MAPK signaling pathway.