Dandelion sterol improves diabetes mellitus-induced renal injury in in vitro and in vivo study

蒲公英甾醇在体外和体内研究中均能改善糖尿病引起的肾损伤

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Abstract

The purpose of our research was to evaluate Dandelion sterol's treatment effects on diabetes mellitus-induced renal injury in in vitro and in vivo study. The rats were divided into five groups as normal control (Ctrl), diabetic nephropathy model (Model), Dandelion sterol low-dose treated (Dan-Low), Dandelion sterol middle-dose treated (Dan-Middle), and Dandelion sterol high-dose treated (Dan-High). Measuring serum TNF-α, IL-1β, and IL-6 concentrations by Elisa assay, evaluate kidney pathology by HE staining, kidney cell apoptosis of TUNEL, TLR4, and NF-κB(p65) proteins expression by IHC assay, and relative gene expressions by RT-qPCR assay. In the following step, using HK-2 treated with high glucose to model DN cell model to discuss the relative mechanisms, evaluate TNF-α, IL-1β, and IL-6 concentrations by Elisa assay, evaluate cell apoptosis by flow cytometry, evaluate TLR4 and NF-κB(p65) proteins expression by WB assay, relative gene expression by RT-qPCR assay, and NF-κB(p65) nuclear volume by cellular immunofluorescence. Compared with Ctrl group, TNF-α, IL-1β, and IL-6 concentrations and apoptosis cell number were significantly increased, TLR4/NF-κB(p65) pathway was significantly stimulated in Model rats and cell groups. With Dan supplement, the diabetic-induced renal injury was significantly improved (p < .05, respectively). By cell experiment, Dan improved cell apoptosis and inflammatory factors via miR-140-5p. Dan improved diabetes mellitus-induced renal injury via regulation of miR-140-5p/TLR4 axis in in vitro and in vivo study.

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