Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Diabetes. Nov 15, 2023; 14(11): 1672-1692
Published online Nov 15, 2023. doi: 10.4239/wjd.v14.i11.1672
Exploring the targets and molecular mechanism of glycyrrhetinic acid against diabetic nephropathy based on network pharmacology and molecular docking
Fan-Di Meng, Ling Yuan, Duo-Jie Xu, Meng-Ying Che, Shao-Zhang Hou, Dou-Dou Lu, Wen-Jing Liu, Yi Nan
Fan-Di Meng, Duo-Jie Xu, Meng-Ying Che, Dou-Dou Lu, Yi Nan, Traditional Chinese Medicine College, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Fan-Di Meng, Wen-Jing Liu, Yi Nan, Key Laboratory of Ningxia Ethnomedicine Modernization, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Ling Yuan, College of Pharmacy, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Shao-Zhang Hou, Ningxia Key Laboratory of Craniocerebral Diseases, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Author contributions: Meng FD, Yuan L, Xu DJ, Che MY, Hou SZ, and Lu DD designed the study; Meng FD, Yuan L, Lu DD, and Yi Nan conducted the study; Che MY and Hou SZ contributed new reagents and analytical tools; Meng FD, Yuan L, and Xu DJ analyzed the data and wrote the manuscript; and all authors have read and approved the final manuscript.
Supported by Ningxia Natural Science Foundation, No. 2022AAC02039; National Natural Science Foundation of China, No. 81860894, 82260879, and 81674096; Ningxia Innovation Team of the Foundation and Clinical Researches of Diabetes and its Complications, No. NXKJT2019010.
Institutional review board statement: The study was reviewed and approved by the Medical Ethics Review Committee of Ningxia Medical University (No. 2018-144).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: Technical appendix, statistical code, and dataset available from the corresponding author at 20080011@nxmu.edu.cn. Participants gave informed consent for data sharing.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Yi Nan, MD, PhD, Professor, Traditional Chinese Medicine College, Ningxia Medical University, No. 1160 Shengli Street, Yinchuan 750004, Ningxia Hui Autonomous Region, China. 20080011@nxmu.edu.cn
Received: August 23, 2023
Peer-review started: August 23, 2023
First decision: September 14, 2023
Revised: September 25, 2023
Accepted: October 23, 2023
Article in press: October 23, 2023
Published online: November 15, 2023
Processing time: 78 Days and 17.2 Hours
Abstract
BACKGROUND

Diabetic nephropathy (DN) stands as the most prevalent chronic microvascular complication of diabetes mellitus. Approximately 50% of DN patients progress to end-stage renal disease, posing a substantial health burden.

AIM

To employ network pharmacology and molecular docking methods to predict the mechanism by which glycyrrhetinic acid (GA) treats DN, subsequently validating these predictions through experimental means.

METHODS

The study initially identified GA targets using Pharm Mapper and the TCMSP database. Targets relevant to DN were obtained from the Genecards, OMIM, and TTD databases. The Venny database facilitated the acquisition of intersecting targets between GA and DN. The String database was used to construct a protein interaction network, while DAVID database was used to conducted Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Ontology (GO) analysis. Molecular docking experiments were performed using Autodock software with selected proteins. Experimental validation was conducted using renal proximal tubular cells (HK-2) as the study subjects. A hyperglycemic environment was simulated using glucose solution, and the effect of GA on cell viability was assessed through the cell counting kit-8 method. Flow cytometry was employed to detect cell cycle and apoptosis, and protein immunoblot (western blot) was used to measure the expression of proteins of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway and insulin resistance pathway, including insulin receptor (INSR), PI3K, p-PI3K, AKT, p-AKT, and glycogen synthase kinase-3 (GSK3).

RESULTS

A total of 186 intersecting targets between GA and DN were identified, which were associated with 144 KEGG-related enrichment pathways, 375 GO biological process entries, 45 GO cellular component entries, and 112 GO cellular function entries. Molecular docking demonstrated strong binding of GA to mitogen-activated protein kinase (MAPK)-1, SRC, PIK3R1, HSP90AA1, CASPASE9, HARS, KRAS, and MAPK14. In vitro experiments revealed that GA inhibited HK-2 cell viability, induced cell cycle arrest at the G2/M phase, and reduced apoptosis with increasing drug concentration. Western blot analysis showed that GA differentially up-regulated GSK3 protein expression, up-regulated AKT/p-AKT expression, down-regulated INSR, AKT, p-AKT, PI3K, and p-PI3K protein expression, and reduced p-PI3K/PI3K levels under high glucose conditions.

CONCLUSION

GA may protect renal intrinsic cells by modulating the PI3K/AKT signaling pathway, thereby inhibiting HK-2 cell viability, reducing HK-2 cell apoptosis, and inducing cell cycle arrest at the G0/G1 phase.

Keywords: Network pharmacology, Molecular docking, Diabetic nephropathy, Glycyrrhetinic acid, Mechanism of action

Core Tip: Diabetes nephropathy (DN) brings a huge burden to human health. Through network pharmacology, we found that glycyrrhetinic acid (GA) has a therapeutic effect on DN, and found 186 therapeutic targets. We speculated and verified that GA plays a role in treating DN by regulating phosphatidylinositol 3-kinase/protein kinase B signaling pathway, inhibiting the proliferation of HK-2 cells, blocking the cell cycle in the G2/M phase, and reducing apoptosis of HK-2 cells. This study provides a new development direction for the treatment of DN.