基于网络药理学、分子对接和细胞试验探究麻杏石甘汤对牛支原体感染的治疗机制

doi:10.16431/j.cnki.1671-7236.2026.02.043

Abstract

Abstract:

Objective Through the use of network pharmacology， molecular docking， and cell experiments， this study aimed to explore the therapeutic mechanism of Maxing Shigan decoction against Mycoplasma bovis infection， providing scientific basis for the modernization of traditional Chinese medicine and new strategies for the treatment of Mycoplasma bovis infection. Method Relying on bioinformatics platforms such as TCMSP， PubChem and SwissTargetPrediction， the active ingredients of the constituent herbs in Maxing Shigan decoction and their related targets were screened. The targets of Mycoplasma bovis related diseases were obtained through GeneCards and OMIM databases. The STRING database was used to construct protein-protein interaction network， and Cytoscape 3.9.1 software was used for network topology analysis. Meanwhile， the DAVID platform was used for GO function and KEGG pathway enrichment analysis. Molecular docking and visualization were completed using PyMOL 2.4.0 and AutoDockTool 1.5.6 software. MDBK cells were divided into blank control group， model group， and low， medium， and high dose groups （2.5， 5 and 10 mg/mL） of Maxing Shigan decoction. Except for blank control group， all other groups were infected with Mycoplasma bovis PG45 with a multiplicity of infection of 1 000 for 4 h. After cleaning， they were replaced with drug containing medium or complete medium， and continued to be cultured for 24 h. Real-time quantitative PCR was used to detect the Ct value and the mRNA expression of relevant inflammatory factors in each group of cells. Result Network pharmacology analysis revealed that Maxing Shigan decoction contained 134 active ingredients， and there were 93 targets related to Mycoplasma bovis-related diseases. A total of 20 intersection targets were identified between them， among which interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， matrix metallo proteinase 9（MMP9）， caspase 3 （CASP3） and prostaglandin peroxide synthase 2 （PTGS2） were the key core targets. GO function enrichment analysis mainly involved positive regulation of transcription by RNA polymerase Ⅱ， extracellular space， cytoplasm， nucleus， cytokine activity， etc. KEGG pathway enrichment analysis mainly covered signaling pathways related to IL-17， TNF， mitogen-activated protein kinase （MAPK），etc. The molecular docking results showed that the binding energy of the active ingredients of Maxing Shigan decoction was less than -17.8 kJ/mol， and had good binding ability. Among them， quercetin had the smallest binding energy to CASP3， which was -28.87 kJ/mol. Cell experiments showed that compared with model group， the Ct values were significantly increased （P<0.05）， the expression of IL-2 and IL-10 genes were significantly upregulated （P<0.05）， while those of IL-6， IL-8， TNF-α， IL-17A，CASP3， MMP9 and PTGS2 genes were significantly downregulated （P<0.05）， in each dose group of Maxing Shigan decoction. Conclusion Through network pharmacology prediction and cell experimental validation， this study revealed that Maxing Shigan decoction exered anti-Mycoplasma bovis infection effects by acting on targets such as IL-6， TNF and MMP9， regulating the secretion of inflammatory factors， and inhibiting inflammatory responses.

Key words: Maxing Shigan decoction; Mycoplasma bovis; network pharmacology; molecular docking

CLC Number:

S853.74

YU Zhaizhuo, ZHANG Liang, CHEN Jiu, WU Zhiyong, ZHU Yangdong, YAN Bin, YANG Hongjun. Exploring the Therapeutic Mechanism of Maxing Shigan Decoction Against Mycoplasma bovis Infection Through Network Pharmacology， Molecular Docking， and Cellular Experiments[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(2): 995-1008.

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基因 Genes	引物序列 Primer sequences （5'→3'）		退火温度 Annealing temperature/℃	GenBank登录号 GenBank accession No.
TNF-α	F： CGGTGGTGGGACTCGTATG	R： GCTGGTTGTCTTCCAGCTTCA	60	NM_173966.3
IL-6	F： TCTGGGTTCAATCAGGCGAT	R： GTCAGTGTTTGTGGCTGGAG	56	NM_173923.2
IL-8	F： TGGGCCACACTGTGAAAAT	R： TCATGGATCTTGCTTCTCAGC	57	NM_173925.2
IL-10	F： TGTTGACCCAGTCTCTGCTG	R： GGCATCACCTCTTCCAGGTA	57	NM_174088.1
IL-17A	F： GAACTTCATCTATGTCACTGC	R： TGGACTCTGTGGGATGATGA	54	NM_001008412.2
IL-2	F： AACCTCAACTCCTGCCACAA	R： TGTGTTCCCCGTAGAGCTTG	56	NM_180997.2
CASP3	F：AGAACTTGGTGGTCCATACA	R：CTCAACCCGTCTCCCTTTAT	54	NM_001077840.1
MMP9	F：GGAGAACCACGAACCAACCT	R：CCAGGAGTGTAGCCATAGCG	58	NM_174744.2
PTGS2	F：GCATAAGCTGCGCCTTTTCA	R：TCTGTTGTGTTCCCGTAGCC	56	NM_174445.2
β-actin	F： TGCTGTCACCTTCACCGTTC	R： CTGTTAGCTGCGTTACACCCT	57	NM_173979.3

No.	靶点 Targets	度值 Degree	介数中心性 Betweenness centrality	接近中心性 Closeness centrality
1	IL-6	17	26.356	0.947
2	TNF	17	26.356	0.947
3	MMP9	16	18.356	0.900
4	CASP3	15	12.489	0.857
5	PTGS2	15	12.489	0.857
6	STAT3	14	7.180	0.818
7	PPARG	13	35.765	0.782
8	IL-2	13	5.275	0.782
9	NOS2	11	1.698	0.720
10	ARG1	11	3.309	0.720

靶点 Targets	结合能Binding energy
靶点 Targets	柚皮素 Naringenin	山柰酚 Kaempferol	槲皮素 Quercetin
IL-6	－21.25	－24.43	－24.31
TNF	－18.12	－19.83	－20.55
MMP9	－17.86	－23.22	－17.95
PTGS2	－22.09	－27.91	－28.45
CASP3	－23.26	－26.86	－28.87
STAT3	－22.81	－22.09	－21.63

Exploring the Therapeutic Mechanism of Maxing Shigan Decoction Against Mycoplasma bovis Infection Through Network Pharmacology， Molecular Docking， and Cellular Experiments

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