Mechanism underlying efficacy of Shugan Sanjie decoction (疏肝散结汤) on plasma cell mastitis, based on network pharmacology and experimental verification

doi:10.19852/j.cnki.jtcm.20220311.004

Abstract

Abstract:

OBJECTIVE: To investigate the mechanism underpinning the effeicay of Shugan Sanjie decoction (疏肝散结汤, SGSJD) on plasma cell mastitis (PCM) based on network pharmacology, and to verify it through in vitro.

METHODS: Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine were used to screen effective compounds and drug targets; Online Mendelian Inheritance in Man and GeneCards were used to search for PCM targets. The potential targets of SGSJD in treating PCM were obtained after the drug targets and disease targets were crossed. Cytoscape software was used to establish and analyze the network of Chinese medicines-active compounds-targets-disease; STRING database platform was used to analyze Protein Protein Interaction network; Bioconductor software package was used to perform Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment for potential targets. Western blot analysis was used to verify the janus kinase/signal transducer and activator of transcription (JAK-STAT) pathway in vitro.

RESULTS: (a) 47 potential pharmacological components of SGSJD treatment of PCM were screened including quercetin, luteolin, kaempferol and others; 20 common targets were obtained, including interleukin-6 (IL-6), epidermal growth factor receptor, estrogen receptor 1, nitric oxide synthase 3 and others; a number of signal pathways were available, of which advanced glycation end product/ receptor for advanced glycation end products signaling pathway in diabetic complications, hypoxia-inducible factor 1 signaling pathway and janus tyrosine kinase-signal transducer and transcription activator (JAK-STAT) signaling pathway were the main signal pathways related to PCM. (b) Compared with the Blank group, the expressions of p-JAK2/JAK2, p-STAT3/STAT3 and IL-6 protein in the Model group were significantly increased (P < 0.01); Compared with the Model group, the expression of p-JAK2/JAK2, p-STAT3/STAT3, and IL-6 protein in the treatment group were significantly reduced in a dose-dependent manner (P < 0.05). Compared with the Model group, the dexamethasone significantly reduced the expression of p-JAK2/JAK2, p-STAT3/STAT3, and IL-6 (P < 0.01).

CONCLUSIONS: The SGSJD may regulate the JAK-STAT signaling pathway to achieve the effect of treating PCM by reducing the expression of p-JAK2/JAK2, p-STAT3/STAT3 and IL-6 in a dose-dependent manner.

Key words: plasma cells, mastitis, network pharmacology, molecular mechanism, Shugan Sanjie decoction

QIAO Nan, WANG Qinnan, MA Chaoqun, CHEN Dexuan, CHEN Haidong, LU Yaoyao. Mechanism underlying efficacy of Shugan Sanjie decoction (疏肝散结汤) on plasma cell mastitis, based on network pharmacology and experimental verification[J]. Journal of Traditional Chinese Medicine, 2022, 42(3): 400-407.

Figures/Tables 8

Figure 1 Targets of drugs and disease

Table 1 Active components of SGSJD

TCMSP Number	Components	TCMSP Number	Components
MOL000449	Stigmasterol	MOL003283	(2R,3R,4S)-4-(4-Hydroxy-3-Methoxy-Phenyl)-7-Methoxy-2,3-Dimethylol-Tetralin-6-Ol
MOL000354	Isorhamnetin	MOL003290	(3R,4R)-3,4-Bis[(3,4-Dimethoxyphenyl)Methyl]Oxolan-2-One
MOL000422	Kaempferol	MOL003315	3beta-Acetyl-20,25-Epoxydammarane-24alpha-Ol
MOL004598	3,5,6,7-Tetramethoxy-2-(3,4,5-Trimethoxyphenyl) Chromone	MOL000211	Mairin
MOL004718	Α-Spinasterol	MOL003347	Hyperforin
MOL000098	Quercetin	MOL000522	Arctiin
MOL000282	Ergosta-7,22E-Dien-3beta-Ol	MOL000006	Luteolin
MOL000283	Ergosterol Peroxide	MOL003044	Chryseriol
MOL000296	Hederagenin	MOL003542	8-Isopentenyl-Kaempferol
MOL004355	Spinasterol	MOL004074	Stigmasterol Glucoside_Qt
MOL006756	Schottenol	MOL001406	Crocetin
MOL007165	10α-Cucurbita-5,24-Diene-3β-Ol	MOL004561	Sudan III
MOL007171	5-Dehydrokarounidiol	MOL003095	5-Hydroxy-7-Methoxy-2-(3,4,5-Trimethoxyphenyl)Chromone
MOL001689	Acacetin	MOL000797	(22e,24r)-Ergosta-7,22-Dien-3-One
MOL000173	Wogonin	MOL000798	Ergosta-7,22-Diene-3β-Ol
MOL000228	(2R)-7-Hydroxy-5-Methoxy-2-Phenylchroman-4-One	MOL000816	Ergosta-7,22-Dien-3-One
MOL002714	Baicalein	MOL000817	Ergosta-5,7,22-Trien-3-Ol
MOL002928	Oroxylin A	MOL004085	Taraxasterol
MOL002933	5,7,4'-Trihydroxy-8-Methoxyflavone	MOL000394	Choline
MOL002934	Neobaicalein	MOL003837	Esculetin
MOL000358	Beta-Sitosterol	MOL000414	Caffeic Acid
MOL000359	Sitosterol	MOL006554	Taraxerol
MOL000073	Ent-Epicatechin
MOL001458	Coptisine
MOL002897	Epiberberine

Figure 2 Network of herbal medicine-component-target-disease CRP: c-reactive protein; DHFR: dihydrofolate reductase; EGFR: epidermal growth factor receptor; ERBB2: erb-b2 receptor tyrosine kinase 2; ESR1: estrogen receptor 1; ICAM1: intercellular adhesion molecule 1; IFNG: interferon gamma; IL6: interleukin 6; IRF1: interferon regulatory factor 1; MCL1: myeloid cell leukemia sequence 1; MUC1: Mucin 1; NOS3: nitric oxide synthase 3; NR3C1: nuclear receptor subfamily 3 group c member 1; PGR: progesterone receptor; RASA1: ras p21 protein activator 1; SELE: selectin e; SRD5A1: steroid 5 alpha-reductase 1; TP63: tumor protein p63; VCAM1: vascular cell adhesion molecule 1; ADH1B: alcohol dehydrogenase 1b (class i), beta polypeptide. CH: Chaihu (Radix Bupleuri Chinensis); FL: Fuling (Poria); GL: Gualou (Fructus et Semen Trichosanthis); HQ: Huangqin (Radix Scutellariae Baicalensis); LQ: Lianqiao (Fructus Forsythiae Suspensae); PGY: Pugongying (Herba Taraxaci Mongolici); XF: Xiangfu (Rhizoma Cyperi); ZL: Zhuling (Polyporus); ZZ: Zhizi (Fructus Gardeniae).

Figure 3 Network of protein protein interaction RASA1: ras p21 protein activator 1; MCL1: myeloid cell leukemia sequence 1; ERBB2: erb-b2 receptor tyrosine kinase 2; TP63: tumor protein p63; SRD5A1: steroid 5 alpha-reductase 1; ICAM1: intercellular adhesion molecule 1; IL6: interleukin 6; EGFR: epidermal growth factor receptor; DHFR: dihydrofolate reductase; IFNG: interferon gamma; ESR1: estrogen receptor 1; PGR: progesterone receptor; MUC1: Mucin 1; NOS3: nitric oxide synthase 3; SELE: selectin e; IRF1: interferon regulatory factor 1; VCAM1: vascular cell adhesion molecule 1; NR3C1: nuclear receptor subfamily 3 group c member 1; CRP: c-reactive protein.

Figure 4 Ranking of key targets EGFR: epidermal growth factor receptor; IL6: interleukin 6; ERBB2: erb-b2 receptor tyrosine kinase 2; ESR1: estrogen receptor 1; ICAM1: intercellular adhesion molecule 1; IFNG: interferon gamma; NOS3: nitric oxide synthase 3; CRP: c-reactive protein; MUC1: Mucin 1; VCAM1: vascular cell adhesion molecule 1; NR3C1: nuclear receptor subfamily 3 group c member 1; SELE: selectin e; IRF1: interferon regulatory factor 1; PGR: progesterone receptor; MCL1: myeloid cell leukemia sequence 1; RASA1: ras p21 protein activator 1.

Figure 5 Analysis of GO function DNA: deoxyribonucleic acid; RNA: ribonucleic acid;NADP: nicotinamide adenine dinucleotide phosphate; NADPH: nicotinamide adenine dinucleotide phosphate (reduced state).

Figure 6 Analysis of KEGG pathway enrichment KEGG：Kyoto encyclopedia of genes and genomes; AGE-RAGE: advanced glycation end products/receptor for advanced glycation end products; HIF-1: hypoxia inducible factor 1; TNF: tumor necrosis factor; JAK-STAT: janus kinase/signal transducer and activator of transcription; EGFR: epidermal growth factor receptor; PI3K-Akt: phosphatidylinositol 3-kinase/Akt.

Figure 7 Western blot analyses A: Western blot picture. Bl: culture solution. Mo: culture solution and LPS, 1.0 μg/mL; Lo: LPS, 1.0 μg/mL and SGSJD extract, 250 μg/mL; Me: LPS, 1.0 μg/mL and SGSJD extract, 500 μg/mL; Hi: LPS, 1.0 μg/mL and SGSJD extract, 750 μg/mL; Po: dexamethasone, 50 μg/mL. B: comparison of the relative content of IL-6 in each group. C: comparison of relative expression of JAK2. D: comparison of relative expression of STAT3. LPS: lipopolysaccharide; Bl: blank group; Mo: model group; Lo: low group; Me: medium group; Hi: high group; Po: positive group; SGSJD: Shugan Sanjie decoction; JAK: janus kinase. Compared with Blank, aP < 0.01; compared with Model, cP < 0.01, bP < 0.05.

References 16

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