Diabetes, Metabolic Syndrome and Obesity (Oct 2020)
Standard Puerarin Prevents Diabetic Renal Damage by Inhibiting miRNA-140-5p Expression
Abstract
Yanmei Xu, Yan Xiong, Chen Xu, Chuanwen Xu Department of Nephrology, Wuhan Fourth Hospital, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan City, Hubei Province 430033, People’s Republic of ChinaCorrespondence: Chuanwen XuDepartment of Nephrology, Wuhan Fourth Hospital, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 473 Hanzheng Street, Qiaokou District, Wuhan City, Hubei Province 430033, People’s Republic of ChinaTel +86-18607171819Email [email protected]: This study was designed to use in vivo and in vitro approaches to evaluate puerarin in diabetes-induced renal injury.Materials and Methods: SD rats were divided into NC (normal control), Model (diabetic induced renal injury model), SP-L (model rats treated with low-dose standard puerarin), SP-M (model rats treated with middle-dose standard puerarin), and SP-H (model rats treated with high-dose standard puerarin) groups. We evaluated fasting blood-glucose (FBG), urinary albumin/creatinine ratio (UACR), body weight, and kidney index (KI) in the different groups. TNF-α, IL-1β, and IL-6 concentrations were measured using Elisa assays. HE staining and TUNEL assays were used to evaluate pathology and apoptosis in kidney tissues, respectively. Relative gene and protein expression was measured using RT-qPCR and Western blot assays. Apoptosis was measured using flow cytometry. The correlation between miRNA-145-5p and TLR4 was assessed using dual-luciferase reporter gene assays.Results: The pathology and apoptosis cell number were deteriorate in Model group; TNF-α, IL-1β and IL-6 concentrations, FGB, UACR and KI were increased and body weight was depressed; meanwhile, relative gene and proteins expressions (miRNA-145-5p, TLR4, MyD88 and NF-κB p65) were significantly different in Model group in vivo and vitro study compared with NC group. SP treatment significantly improved the pathology and apoptosis levels in the tissues, as well as TNF-α, IL-1β and IL-6 concentrations, FGB, UACR, body weight, and KI. In vitro cell studies revealed that SP could prevent renal injury induced by diabetes through the miRNA-145-5p/TLR4 axis.Conclusion: SP prevents diabetes-induced renal damage via miRNA-145-5p overexpression and reduces TLR4/MyD88/NF-κB (p65) pathway activation in vitro and in vivo.Keywords: SP, renal injury, apoptosis, miRNA-145-5p, TLR4/MyD88/NF-κB (p65)
