Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (1): 110-118.DOI: 10.19852/j.cnki.jtcm.2026.01.010
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Network pharmacological and experimental validation of the mechanism of Chaihu Guizhi Ganjiang decoction (柴胡桂枝干姜汤) regulating T helper cell 17/regulatory T cell balance to improve autoimmune hepatitis
LIANG Zihao1,2, GAO Jie1,2, SONG Jinyun1,2, ZHENG Qin2(
), ZHAO Hongyu1,2()
- 1 Clinical Research Center, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210023, China
2 Department of Oncology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210023, China
-
Received:2024-10-14Accepted:2025-05-29Online:2026-02-15Published:2026-01-28 -
Contact:ZHAO Hongyu, Clinical Research Center, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210023, China; Department of Oncology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210023, China.njdie001@njucm.edu.cn ; ZHENG Qin, Department of Oncology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210023, China.njyy040@njucm.edu.cn ;Telephone: +86-13814065775 -
Supported by:Nanjing Health Science and Technology Key Medical Science and Technology Development Program: Mechanism of Action of the Modified Si-Miao Powder with Sanguisorba Carbonisata in Regulating Microbiota and Microecology for the Treatment of Recurrent Vulvovaginal Candidiasis(ZKX22039)
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LIANG Zihao, GAO Jie, SONG Jinyun, ZHENG Qin, ZHAO Hongyu. Network pharmacological and experimental validation of the mechanism of Chaihu Guizhi Ganjiang decoction (柴胡桂枝干姜汤) regulating T helper cell 17/regulatory T cell balance to improve autoimmune hepatitis[J]. Journal of Traditional Chinese Medicine, 2026, 46(1): 110-118.
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Figure 1 Venn diagram of the AIH and CGGD target genes ETCM: Encyclopedia of Traditional Chinese Medicine; TCMSP: Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform; AIH: autoimmune hepatitis; CGGD: Chaihu Guizhi Ganjiang decoction.
Figure 1 Venn diagram of the AIH and CGGD target genes ETCM: Encyclopedia of Traditional Chinese Medicine; TCMSP: Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform; AIH: autoimmune hepatitis; CGGD: Chaihu Guizhi Ganjiang decoction.
Figure 2 Enrichment analysis of common target genes of AIH and CGGD A: top five significantly enriched terms (P < 0.05) in the biological process (red), cellular component (green), and molecular function (purple) categories according to GO analysis; B: the top 15 significantly enriched KEGG pathways (P < 0.05). GO: Gene Ontology; NMDA: N-methyl-D-aspartic acid; PI3K: phosphoinositide 3-kinase; AKT: protein kinase B; cAMP: cyclic adenosine monophosphate; Rap 1: ras-proximate-1; NOD: nucleotide-binding Oligomerization; MAPK: mitogen-activated protein kinase; NF-kapper B: nuclear factor kappa-B; Th17: T helper cell 17; AIH: autoimmune hepatitis; CGGD: Chaihu Guizhi Ganjiang decoction; KEGG: kyoto encyclopedia of genes and genomes.
Figure 2 Enrichment analysis of common target genes of AIH and CGGD A: top five significantly enriched terms (P < 0.05) in the biological process (red), cellular component (green), and molecular function (purple) categories according to GO analysis; B: the top 15 significantly enriched KEGG pathways (P < 0.05). GO: Gene Ontology; NMDA: N-methyl-D-aspartic acid; PI3K: phosphoinositide 3-kinase; AKT: protein kinase B; cAMP: cyclic adenosine monophosphate; Rap 1: ras-proximate-1; NOD: nucleotide-binding Oligomerization; MAPK: mitogen-activated protein kinase; NF-kapper B: nuclear factor kappa-B; Th17: T helper cell 17; AIH: autoimmune hepatitis; CGGD: Chaihu Guizhi Ganjiang decoction; KEGG: kyoto encyclopedia of genes and genomes.
Figure 3 CGGD alleviates ConA-induced liver injury A: histological changes in the liver were assessed using HE staining. Scale bars = 100 μm; A1: Ctrl group; A2: ConA group; A3: CGGD-L group; A4: CGGD-M group; A5: CGGD-H group; B: activities of and AST; C: activities of ALT. Ctrl Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of 10 μL of normal saline via the tail vein on the eighth day; ConA Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of ConA at a dose of 20 mg/kg according to body weight on the eighth day; CGGD-L Group: received ConA by gavage at doses of 2000, mg/kg once daily for one week prior to model induction; CGGD-M Group: received ConA by gavage at doses of 4000 mg/kg once daily for one week prior to model induction; CGGD-H Group: received ConA by gavage at doses of 8000 mg/kg once daily for one week prior to model induction. AST: aspartate aminotransferase; ALT: alanine aminotransferase; CGGD: Chaihu Guizhi Ganjiang decoction. Statistical differences between groups were analyzed using a one-way analysis of variance. Data were presented as mean ± standard error of the mean (n = 6). Compared with the Ctrl group, aP < 0.01; compared with the ConA group, bP < 0.01; compared with the CGGD-L group, cP < 0.01; compared with the CGGD-M group, dP < 0.01.
Figure 3 CGGD alleviates ConA-induced liver injury A: histological changes in the liver were assessed using HE staining. Scale bars = 100 μm; A1: Ctrl group; A2: ConA group; A3: CGGD-L group; A4: CGGD-M group; A5: CGGD-H group; B: activities of and AST; C: activities of ALT. Ctrl Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of 10 μL of normal saline via the tail vein on the eighth day; ConA Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of ConA at a dose of 20 mg/kg according to body weight on the eighth day; CGGD-L Group: received ConA by gavage at doses of 2000, mg/kg once daily for one week prior to model induction; CGGD-M Group: received ConA by gavage at doses of 4000 mg/kg once daily for one week prior to model induction; CGGD-H Group: received ConA by gavage at doses of 8000 mg/kg once daily for one week prior to model induction. AST: aspartate aminotransferase; ALT: alanine aminotransferase; CGGD: Chaihu Guizhi Ganjiang decoction. Statistical differences between groups were analyzed using a one-way analysis of variance. Data were presented as mean ± standard error of the mean (n = 6). Compared with the Ctrl group, aP < 0.01; compared with the ConA group, bP < 0.01; compared with the CGGD-L group, cP < 0.01; compared with the CGGD-M group, dP < 0.01.
Table 1 mRNA expression levels of cytokine levels
| Group | n | IL-1β | IL-6 | TNF-α | IL-10 |
|---|---|---|---|---|---|
| Ctrl | 6 | 1.00±0.46a | 1.00±0.30a | 1.00±0.22a | 1.00±0.32c |
| ConA | 6 | 27.22±5.50 | 32.98±4.95 | 17.56±3.34 | 2.08±0.39 |
| GCCD-L | 6 | 16.67±4.21a | 25.44±2.26b | 8.82±1.93a | 2.23±0.51 |
| GCCD-M | 6 | 13.16±3.05a | 19.32±2.82a | 6.57±1.01a | 2.41±0.55 |
| GCCD-H | 6 | 5.43±2.06a | 13.99±2.95a | 2.83±0.90a | 4.13±0.91a |
Table 1 mRNA expression levels of cytokine levels
| Group | n | IL-1β | IL-6 | TNF-α | IL-10 |
|---|---|---|---|---|---|
| Ctrl | 6 | 1.00±0.46a | 1.00±0.30a | 1.00±0.22a | 1.00±0.32c |
| ConA | 6 | 27.22±5.50 | 32.98±4.95 | 17.56±3.34 | 2.08±0.39 |
| GCCD-L | 6 | 16.67±4.21a | 25.44±2.26b | 8.82±1.93a | 2.23±0.51 |
| GCCD-M | 6 | 13.16±3.05a | 19.32±2.82a | 6.57±1.01a | 2.41±0.55 |
| GCCD-H | 6 | 5.43±2.06a | 13.99±2.95a | 2.83±0.90a | 4.13±0.91a |
Figure 4 CGGD regulates Th17/Treg homeostasis in ConA-induced AIH in mice A: flow cytometry analysis of CD4+IL-17+ (Th17) in mouse spleen; A1: Ctrl group; A2: ConA group; A3: CGGD-L group; A4: CGGD-M group; A5: CGGD-H group; A6: the quantitative frequency analysis of Th17 cells; B: flow cytometry analysis of CD4+Foxp3+ (Treg) cells in mouse spleen; B1: Ctrl group; B2: ConA group; B3: CGGD-L group; B4: CGGD-M group; B5: CGGD-H group; B6: the quantitative frequency analysis of Treg cells. Control Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of 10 μL of normal saline via the tail vein on the eighth day; ConA Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of ConA at a dose of 20 mg/kg according to body weight on the eighth day; CGGD-L, CGGD-M & CGGD-H Group: received ConA by gavage at doses of 2000, 4000, and 8000 mg/kg once daily for one week prior to model induction. Th17: T helper cell 17; Treg: regulatory T cell. CGGD: Chaihu Guizhi Ganjiang decoction. Statistical differences between groups were analyzed using a one-way analysis of variance. Data were presented as mean ± standard error of the mean (n = 6). Compared with the Ctrl group, aP < 0.01; compared with the ConA group, bP < 0.01; compared with the CGGD-L group, cP < 0.01; compared with the CGGD-M group, d P < 0.01.
Figure 4 CGGD regulates Th17/Treg homeostasis in ConA-induced AIH in mice A: flow cytometry analysis of CD4+IL-17+ (Th17) in mouse spleen; A1: Ctrl group; A2: ConA group; A3: CGGD-L group; A4: CGGD-M group; A5: CGGD-H group; A6: the quantitative frequency analysis of Th17 cells; B: flow cytometry analysis of CD4+Foxp3+ (Treg) cells in mouse spleen; B1: Ctrl group; B2: ConA group; B3: CGGD-L group; B4: CGGD-M group; B5: CGGD-H group; B6: the quantitative frequency analysis of Treg cells. Control Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of 10 μL of normal saline via the tail vein on the eighth day; ConA Group: 200 μL of normal saline was administered daily for one week, followed by an intravenous injection of ConA at a dose of 20 mg/kg according to body weight on the eighth day; CGGD-L, CGGD-M & CGGD-H Group: received ConA by gavage at doses of 2000, 4000, and 8000 mg/kg once daily for one week prior to model induction. Th17: T helper cell 17; Treg: regulatory T cell. CGGD: Chaihu Guizhi Ganjiang decoction. Statistical differences between groups were analyzed using a one-way analysis of variance. Data were presented as mean ± standard error of the mean (n = 6). Compared with the Ctrl group, aP < 0.01; compared with the ConA group, bP < 0.01; compared with the CGGD-L group, cP < 0.01; compared with the CGGD-M group, d P < 0.01.
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