Dandelion sterol improves diabetes mellitus–induced renal injury in in vitro and in vivo study
2021
Lin Tian | Peng Fu | Min Zhou | Jiping Qi
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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