Abstract

The purpose of this study was to investigate the potential effects of Huaiqihuang (HQH) granule, a Chinese herbal medicine, in treating proteinuria and to reveal its possible mechanism. MPC5 podocytes were cultured in vitro at 37°C and induced with tunicamycin (TM). The TM-induced cells were treated with HQH at different concentrations. The cell proliferation was detected using the MTT assay. The optimal effective dose of HQH for MPC5 cells was determined by the MTT assay and LDH assay respectively. The influences of HQH on the proteinuria-related protein expression and the signaling pathway associated protein expression were also detected using quantitative reverse transcription PCR and Western blotting analysis. The results showed that the MPC5 cell model was successfully constructed in vitro. The HQH application could improve the harmful effects induced by TM on the MPC5 cells, including promoted cell proliferation and suppressed cell apoptosis. Furthermore, the protein expression, including podocin, nephrin, and synaptopodin was down-regulated by the TM treatment in the MPC5 cells. On contrary, the expression of these proteins was up-regulated after the HQH application. Also, the effect of TM on integrin α3 and integrin β1 expressions was also reversed by the HQH treatment. Moreover, the HQH application decreased the expression of p-ERK and DNA-damage-inducible transcript 3 (DDIT3 or CHOP) in the MPC5 cells, which was opposite to the effect observed in the cells treated with TM. Taken together, our study suggest that HQH application may protect podocytes from TM damage by suppressing the p-ERK/CHOP signaling pathway.

Highlights

  • Proteinuria is a typical symptom of chronic kidney disease, which is caused by the dysfunction of glomerulus filtration membrane [1]

  • The Western blotting showed that the expression of Cullin-5, a cell proliferation indicator, was higher in the MPC5 cells treated at 33°C compared to the cells treated at 37°C (Figure 1B)

  • We observed that the protective effect of HQH on the MPC5 cells was more apparent with longer HQH application

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Summary

Introduction

Proteinuria is a typical symptom of chronic kidney disease, which is caused by the dysfunction of glomerulus filtration membrane [1]. Previous studies have demonstrated that the pathological changes in podocytes remain to be the major cause of proteinuria [3,4]. The major treatment methods for proteinuria are medications, including adrenal cortical hormones and immune inhibitors [5,6]. Some of these medications are prohibitive and they may produce large side effects such as high blood pressure, slow growth, diabetes, and femoral head necrosis [7,8]. It is necessary to Submitted: 19 November 2015/Accepted: 08 March 2016 develop useful and inexpensive medications for the proteinuria control

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