Potential application of Nardostachyos Radix et Rhizoma-Rhubarb for the treatment of diabetic kidney disease based on network pharmacology and cell culture experimental verification

ARTICLE HIGHLIGHTS

Research background

Diabetic kidney disease (DKD) is one of the serious complications of diabetes mellitus. It has a poor prognosis and is one of the causes of end-stage renal disease. Existing treatments can improve the symptoms of DKD to some extent. However, they have the disadvantages of side effects and high price.

Research motivation

We performed in vitro cellular experiments to validate the effectiveness of the Nardostachyos Radix et Rhizoma-rhubarb drug pair (NRDP) and to provide new ideas for clinical treatment of DKD.

Research objectives

In this study, we used network pharmacology and molecular docking to predict the targets of NRDP for the treatment of DKD and validated the prediction findings using cellular experiments.

Research methods

Targets for NRDP and DKD were obtained using databases such as TCMSP, Genecards, OMIM, and TTD. Drug-disease intersection targets were obtained based on the VENNY 2.1 database and "drug-component-target-disease" network was constructed. Afterward, Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology enrichment analyses were performed to further observe the relationship between targets and pathways. Finally, molecular docking was performed on the active ingredients of NRDP. Experiments such as the CCK-8 method, flow cytometry, and Western Blot were used to verify the molecular mechanism of NRDP for DKD.

Research results

NRDP may inhibit the viability of high glucose-induced TCMK-1 cells by modulating the advanced glycation end products (AGEs)-receptor for AGEs (RAGE) signaling pathway, thereby blocking cell cycle progression in the G0/G1 phase and reducing apoptosis. It also downregulated the protein expression of p-STAT3, BAX, Caspase3, and Caspase9, and up-regulated the protein levels of BCL-2 and STAT3. These findings verified that NRDP could reduce high glucose-induced TCMK-1 cell injury, thereby restoring their function.

Research conclusions

NRDP may achieve its therapeutic effect on DKD by modulating the AGE-RAGE signaling pathway. NRDP arrests the cell cycle progression at the G0/G1 phase by inhibiting the proliferation of high glucose-induced TCMK-1 cells and reducing their apoptosis. NRDP inhibits the expression of proteins related to the AGE-RAGE signaling pathway in high glucose environment, which delays the progression of DKD.

Research perspectives

We next plan to conduct in vivo animal and omics experiments. To determine the specific components of NRDP in the blood for the treatment of DKD, gene detection will be performed by high-throughput validation methods such as transcriptomics, in order to provide a safe and effective method for clinical treatment of DKD.