Standard Puerarin Prevents Diabetic Renal Damage by Inhibiting miRNA-140-5p Expression

Abstract

Aim

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)

Research Involving Human Participants and/or Animals

All procedures performed in studies involving human participants were performed in accordance with the ethical standards of the institutional and/or national research committee and the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. This study was approved by the ethics committee of Wuhan Fourth Hospital (No.)

Disclosure

The authors declare that they have no conflicts of interest for this work.

Additional information

Funding

There is no funding to report.