Mechanism of Huashi Xingyu Qingre recipe (化湿行淤清热方) in treating oral lichen planus based on network pharmacology and clinical trial verification

doi:10.19852/j.cnki.jtcm.20210707.001

Abstract

Abstract:

OBJECTIVE: To identify the main active components and targets of Huashi Xingyu Qingre recipe (化湿行淤清热方, HXQR) and to investigate its mechanism in the treatment of oral lichen planus (OLP).

METHODS: The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was searched to identify the active ingredients and corresponding targets of HXQR. Disease genes were obtained from the GeneCards database, and a “drug-disease regulatory network” was constructed using Cytoscape software and PERL programming language. The STRING database was used to build a protein-protein interaction network. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) terms were analyzed using R software with a Bioconducter plugin. Finally, the results and the efficacy of HXQR in treating OLP were validated in a clinical trial that included enzyme-linked immunosorbent assay (ELISA) testing and observations of the post-treatment changes in clinical symptoms.

RESULTS: HXQR contained 167 active components and 261 targets, with 391 disease targets. The intersection of these two categories in a Venn diagram revealed 57 drug-disease common targets. A compound-target network was constructed and revealed that the six key pharmaceutical ingredients of HXQR were quercetin, luteolin, wogonin, kaempferol, beta-carotene, and baicalein. The protein-protein interaction network mainly involved core proteins such as ALB, interleukin-6, and AKT1. Drug-disease common targets were enriched in 1628 GO terms and 117 KEGG terms, mainly involving inflammatory responses, viral infections, and tumor-related pathways. ELISA testing indicated that HXQR inhibited the tumor necrosis factor (TNF) signaling pathway by reducing the expression of interleukin-6, matrix metalloproteinase-9, and intercellular adhesion molecule-1. The clinical symptoms of the patients with OLP were significantly improved after 8 weeks of treatment with HXQR.

CONCLUSION: HXQR treats OLP by regulating the TNF signaling pathway, resulting in a marked treatment effect with few adverse effects.

Key words: lichen planus, oral, network pharmacology, clinical trial, Huashi Xingyu Qingre recipe

DENG Qianlan, LU Yueting, YAN Lijuan, LU Hualin, JIN Ruizhe, XU Yanzhi, SONG Jing, LIU Tiejun. Mechanism of Huashi Xingyu Qingre recipe (化湿行淤清热方) in treating oral lichen planus based on network pharmacology and clinical trial verification[J]. Journal of Traditional Chinese Medicine, 2022, 42(2): 304-313.

Figures/Tables 12

Figure 1 Proportions of active components in each ingredient of Huashi Xingyu Qingre recipe

Figure 2 Venn diagram of the active components in each ingredient of Huashi Xingyu Qingre recipe The left bar chart represents each drug ingredient and the number of active components it contains, the top bar represents the number of active components shared by the drug, and the black dot in the middle represents each drug ingredient that contains each active component.

Figure 3 Venn diagram of drug and disease targets The blue part represents 204 drug targets, the yellow part represents 334 disease targets, and the gray part represents 57 drug-disease common targets.

Figure 4 Network of active compounds and common targets The light blue rectangles in the outer circle represent genes. The rectangles in the middle of the circle represent active compounds. The rectangles represent the compounds in Baixianpi (white), Chishao (orange), Fuling (purple), Gancao (green), Honghua (red), Huangbai (brown), Jinyinhua (yellow), Peilan (dark blue), and Yiyiren (pink). The indigo blue rectangles represent common components of two or more ingredients in Huashi Xingyu Qingre recipe.

Figure 5 Protein-Protein Interaction network

Figure 6 Top 20 biological process terms

Figure 7 Top 20 cellular component terms

Figure 8 Top 20 molecular function terms

Figure 9 Top 20 Kyoto Encyclopedia of Genes and Genomes pathways

Figure 10 TNF signaling pathway The red rectangles represent the central nodes in the tumor necrosis factor (TNF) signaling pathway.

Table 1 Pre- and post-treatment serum levels of IL-6, MMP-9, and ICAM-1 ($\bar{x}±s$)

Group	n	IL-6 (pg/mL)	MMP-9 (pg/mL)	ICAM-1 (ng/mL)
Control	28	9.91±1.12	1128.63±30.25	3.97±0.20
Pre-treatment	30	23.79±2.36^a	1634.82±75.60^a	9.03±0.21^a
Post-treatment	30	10.08±1.60^b	1178.03±67.29^b	4.05±0.19^b

Figure 11 Pre- and post-treatment photographs of typical cases of oral lichen planus

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