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. 2022 Jul 20:2022:8336131.
doi: 10.1155/2022/8336131. eCollection 2022.

Mechanisms of Qing-Gan Li-Shui Formulation in Ameliorating Primary Open Angle Glaucoma: An Analysis Based on Network Pharmacology

Lin Mu  1 , Zhiguo Dong  1 , Yinjian Zhang  1
Affiliations

Mechanisms of Qing-Gan Li-Shui Formulation in Ameliorating Primary Open Angle Glaucoma: An Analysis Based on Network Pharmacology

Lin Mu et al. Evid Based Complement Alternat Med. .

Abstract

Objective: In this study, we investigated the mechanism of Qing-Gan Li-Shui formulation (QGLSF) in treating primary open glaucoma (POAG) by network pharmacology and in vitro experiments.

Methods: The active pharmaceutical ingredients (APIs) of GLQSF (prepared with Prunella vulgaris, Kudzu root, Plantago asiatica, and Lycium barbarum) were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Yet Another Traditional Chinese Medicine database (YATCM). The targets of POAG were screened out with GeneCards, OMIM, PharmGKB, Therapeutic Target Database (TTD), and DrugBank databases. The Venny platform was used to summarize the core targets. Topological analysis was performed using Cytoscape3.8.0. A protein-protein interaction network was plotted by STRING online. The key targets were subjected to GO and KEGG enrichment analyses. Finally, the effects of APIs were verified by a model of chloride hexahydrate (CoCl2)-induced retinal ganglion cells-5 (RGC-5).

Results: The main APIs were selected as quercetin (Que) by network pharmacology. Nine clusters of QGLSF targets were obtained by the PPI network analysis, including AKT-1, TP53, and JUN. KEGG enrichment analysis showed that these targets were mainly involved in the AGE-RAGE signaling pathway. By in vitro experiments, Que promoted cell proliferation. The secretion of AKT-1, TP53, JUN, AGE, and RAGE in the cell culture supernatant decreased, as shown by ELISA. The mRNA levels of AKT-1, TP53, JUN, and RAGE decreased, as shown by RT-PCR. QGLSF may employ the AGE-RAGE signaling pathway to counter POAG.

Conclusion: This study preliminarily elucidates the efficacy and mechanism of QGLSF in the treatment of POAG.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Figure 1
Figure 1
Screening of POAG and QGLSF-related targets and APIs. (a) API-target network. Red icons are targets and yellow icons are APIs. (b) Intersections in the venny diagram of POAG-related targets from Genecards, OMIM, PharmGKB, TTD, and Drugbank databases. (c) Intersections in the venny diagram of QGLSF and POAG targets.
Figure 2
Figure 2
The API-target-POAG network. Rectangles represent targets and circles represent active ingredients. Different colors in the circle represent different herbs (red represents Plantago asiatica, purple represents Kudzuvine root, orange represents Lycium barbarum, and green represents Prunella vulgaris).
Figure 3
Figure 3
PPI networks. (a) Initial PPI network; (b) secondary PPI network; (c) PPI network after final screening.
Figure 4
Figure 4
GO and KEGG enrichment analysis (a) GO enrichment analysis (b) KEGG enrichment analysis.
Figure 5
Figure 5
API-target-pathway network (a) API-target-pathway network. Yellow indicates target, green indicates API, and blue indicates pathway. (b) AGE-RAGE signaling pathways.
Figure 6
Figure 6
Cell viability and apoptosis assay. (a) Effect of different concentrations of que on RGC-5 cell viability. (b) Effect of que on CoCl2-inhibited RGC-5 cell viability. (c) Effect of que on CoCl2-induced apoptosis in RGC-5 cells. Values are represented as mean ± S.D. #p < 0.05 versus control group. ∗p < 0.05 versus CoCl2 group. ∗∗p < 0.01 versus CoCl2 group.
Figure 7
Figure 7
Que regulated the expression of hub genes. (a) Expression of RAG in the cell supernatant. (b) Expression of AGEs in the cell supernatant. (c) Expression of AKT-1 in the cell supernatant. (d) Expression of JUN in the cell supernatant. (e) Expression of TP53 in the cell supernatant. Values are represented as mean ± S.D. #p < 0.05 versus control group. ∗p < 0.05 versus CoCl2 group. ∗∗p < 0.01 versus CoCl2 group. ∗∗∗p < 0.001 versus CoCl2 group.
Figure 8
Figure 8
Que regulated the mRNA expression of core targets. (a) mRNA expression of RAG. (b) mRNA expression of TP53. (c) mRNA expression of JUN. (d) mRNA expression of AKT-1. Values are represented as mean ± S.D. #p < 0.05 versus control group. ∗p < 0.05 versus CoCl2 group. ∗∗p < 0.01 versus CoCl2 group.
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