金雀异黄素在甲状旁腺激素诱导的人近曲小管上皮细胞结缔组织生长因子表达中的调控作用

目的 探讨金雀异黄素（Gen）对甲状旁腺激素（PTH）引起的人近曲小管上皮细胞分泌结缔组织生长因子（CTGF）的调控作用。 方法 应用实时定量-聚合酶链反应（real time-PCR）、Western蛋白印迹、报告基因等技术，观察Gen对PTH诱导人近端肾小管上皮细胞系HK-2细胞CTGF表达的影响。使用MAPK通路抑制剂U0126阻断信号通路以明确Gen发挥作用的机制。 结果 HK-2细胞有基础量的CTGF mRNA和蛋白表达，PTH刺激后其表达量显著增加（P ＜ 0.05）。10-10 mol/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，P ＜ 0.01）。Gen剂量依赖性下调PTH诱导的HK-2细胞CTGF表达。正常HK-2细胞有少量磷酸化（p）ERK1/2表达，PTH刺激后p-ERK1/2表达明显升高，以10-10 mol/L PTH作用30 min时效应最强。U0126作用后，CTGF mRNA、蛋白表达均明显下降（P ＜ 0.05）。Gen抑制PTH所致的HK-2细胞ERK1/2活化。 结论 Gen可通过阻断MAPK信号通路抑制PTH诱导的HK-2细胞CTGF表达。

Genistein regulates connective tissue growth factor expression induced by parathyroid hormone in human renal proximal tubular cells

Objective To investigate the role of genistein(Gen) in the expression of connective tissue growth factor (CTGF) induced by parathyroid hormone (PTH) in human renal tubular epithelia cells. Method Real time-PCR, Western blotting, and reporter gene assay were employed to detect the effect of Gen on the expression of CTGF induced by PTH in HK-2 cells. Inhibitor of MAPK signaling pathway was used to ascertain which signal pathway was involved. Results HK-2 cells had basic CTGF mRNA and protein expression, which increased significantly after treatment with PTH. The maximal level of luciferase activity increased 1.96-fold 12 hours after exposure to PTH (10-10 mol/L). Gen dose-dependently decreased the expression of CTGF induced by PTH. Low level of p-ERK1/2 existed in normal HK-2 cells and which increased significantly in response to PTH (10-10 mol/L) and became most remarkable at 30 min. ERK1/2 antagonist U0126 noticeably inhibited the expression of CTGF in HK-2 cells. Gen blunted the phosphorylation of ERK1/2 induced by PTH. Conclusion Gen regulates CTGF expression induced by PTH through inhibiting MAPK signaling pathway in human renal tubular epithelia cells.