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. 2022 Jun;42(3):400-407.
doi: 10.19852/j.cnki.jtcm.20220311.004.

Mechanism underlying efficacy of Shugan Sanjie decoction on plasma cell mastitis, based on network pharmacology and experimental verification

Qiao Nan  1 , Wang Qinnan  2 , M A Chaoqun  3 , Chen Dexuan  1 , Chen Haidong  3 , L U Yaoyao  3
Affiliations

Mechanism underlying efficacy of Shugan Sanjie decoction on plasma cell mastitis, based on network pharmacology and experimental verification

Qiao Nan et al. J Tradit Chin Med. 2022 Jun.

Abstract

Objective: To investigate the mechanism underpinning the effeicay of Shugan Sanjie decoction (, SGSJD) on plasma cell mastitis (PCM) based on network pharmacology, and to verify it through .

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 .

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 ( < 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 ( < 0.05). Compared with the Model group, the dexamethasone significantly reduced the expression of p-JAK2/JAK2, p-STAT3/STAT3, and IL-6 ( < 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.

Keywords: Shugan Sanjie decoction; mastitis; molecular mechanism; network pharmacology; plasma cells.

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Figures

Figure 1
Figure 1. Targets of drugs and disease
Figure 2
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
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
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
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
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
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.
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