Niaoduqing Granules Inhibits TGF-β1-induced Epithelial-Mesenchymal Transition in Human Renal Tubular Epithelial HK-2 Cells

Chuying Huo, Huayi Yang, W. Ning, Lin-zhong Yu, Chunlin Fan, Jingyu Quan, L. Deng, Zhi-Ling Yu, Junshan Liu, Huihui Cao
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Abstract

Background: Chronic renal failure (CRF) is a worldwide public health burden. Niaoduqing granules (NDQ) is widely used for CRF treatment in China. However, the underlying mechanism of NDQ is not fully studied. This study is aimed to investigate whether NDQ ameliorate CRF by inhibiting TGF-β1-induced EMT in human renal tubular epithelial HK-2 cells.Methods: MTT assay and colony formation assay were used to investigate the cytotoxicity of NDQ in HK-2 cells. Morphological changes of HK-2 cells after TGF-β1 or/and NDQ treatment were observed under a microscope. Wound-healing, migration and invasion assays were performed to determine the cell movement, migratory and invasive abilities, respectively. Western blot analysis was carried out to examine the protein levels of TGF-β type I receptor (TβRI) and EMT-associated factors. Fluorescence confocal microscopy was applied to observe the organization of F-actin.Results: NDQ suppressed TβRI expression dose-dependently. NDQ inhibited TGF-β1-stimulated EMT in HK-2 cells, supported by the evidences that NDQ prevented morphology change, attenuated cell migration and invasion, downregulated EMT factors and reorganized F-actin distribution in TGF-β1-stimulated HK-2 cells.Conclusions: NDQ attenuates chronic renal failure which may be associated with inhibition of TβRI expression and EMT process.
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尿毒清颗粒抑制TGF-β1诱导的人肾小管上皮HK-2细胞上皮间质转化
背景:慢性肾衰竭(CRF)是一个世界性的公共卫生负担。脑毒清颗粒(NDQ)在国内被广泛应用于慢性肾功能衰竭的治疗。然而,NDQ的潜在机制尚未得到充分的研究。本研究旨在探讨NDQ是否通过抑制TGF-β1诱导的人肾小管上皮HK-2细胞EMT来改善CRF。方法:采用MTT法和集落形成法观察NDQ对HK-2细胞的细胞毒性。显微镜下观察TGF-β1或/和NDQ处理后HK-2细胞的形态学变化。分别进行伤口愈合、迁移和侵袭实验,以确定细胞的运动、迁移和侵袭能力。Western blot检测TGF-β I型受体(TβRI)蛋白表达水平及emt相关因子水平。采用荧光共聚焦显微镜观察F-actin的组织结构。结果:NDQ抑制TβRI表达呈剂量依赖性。NDQ对TGF-β1刺激的HK-2细胞EMT有抑制作用,证明NDQ能抑制TGF-β1刺激的HK-2细胞的形态学改变,减弱细胞迁移和侵袭,下调EMT因子,重组F-actin分布。结论:NDQ减轻慢性肾功能衰竭可能与抑制TβRI表达和EMT过程有关。
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