Interventional Effects Of Artesunate On The Renal Tubular Epithelial Cells Injury By High Glucose And Its Mechanisms

Objective: To investigate whether artesunate(Art) can inhibit the pro-apoptotic effect of high glucose on rat renal tubular epithelial cells(NRK-52E), and the possible mechanisms in which Art exerts its anti-apoptotic role.Methods : NRK-52 E cells were cultured and divided into six groups: normal control group,high glucose group,Art in low dose group,Art in medium dose group, Art in high dose group, and Enalapril group. After culture for 48 h respectively, MTT assay was used to detect the cell proliferation. The rate of apoptosis was evaluated by flow cytometry AnnexinV-FITC/PI double stains. The protein levels of tumor necrosis factor-alpha(TNF-α), interleukin-8(IL-8) and monocyte chemoattractant protein-1(MCP-1) in the cell culture supernatant were determined using ELISA. The expressions of toll-like receptor-4(TLR4) and nuclear factor-κB(NF-κB) at mRNA and protein levels were detected by RT-PCR and Western blot.Results: ① Compared with the normal control group, high glucose can inhibit NRK-52 E proliferation, induce its apoptosis, and increase the expressions of TNF-α,IL-8 and MCP-1 in the supernatant. The expressions of TLR4 and NF-κB at mRNA and protein in the high glucose group gradually increased(P<0.05). ② Compared with the high glucose group, Art can induce NRK-52 E proliferation, inhibit its apoptosis, and decrease the expressions of TNF-α,IL-8 and MCP-1 in the supernatant. The expressions of TLR4 and NF-κB at mRNA and protein in the Art in low, medium and high dose groups gradually reduced(P<0.05). The interventional effect of Art is shown in dose-dependent manner.Conclusions: Art can inhibit high glucose-induced renal tubular epithelial cells apoptosis. The mechanism of this action probably involves that Art blocks TLR4/NF-κB signaling pathway, reduce expressions of TLR4, NF-κB and inflammatory factors such as TNF-α, IL-8 and MCP-1 in NRK-52 E cells. The anti-inflammatory action and immune regulation of Art could be a novel approach of treating the diabetic nephropathy.