Atractylenolide I ameliorates post-infectious irritable bowel syndrome by inhibiting the polymerase I and transcript release factor and c-Jun N-terminal kinase/inducible nitric oxide synthase pathway

doi:10.19852/j.cnki.jtcm.2025.01.006

Abstract

Abstract:

OBJECTIVE: To explore the therapeutic effect and target of atractylenolide I (AT-I) on post-infectious irritable bowel syndrome (PI-IBS) rats.

METHODS: Therefore, the preliminarily mechanism of AT-I in anti-PI-IBS were first predicted by network pharmacology and molecular docking, then the possible signaling pathways were systematically analyzed. Finally, the potential therapeutic targets and possible signaling pathways of AT-I on PI-IBS in Sprague-Dawley (SD) rat model were verified by experiments.

RESULTS: AT-I could alleviate PI-IBS symptoms and reduce the expression of tumor necrosis factor α, interleukin-6 and Interferon-gamma in PI-IBS SD rat model and inhibit the c-Jun N-terminal kinase/inducible nitric oxide synthase (JNK/iNOS) pathway. Notably, AT-I treatment could inhibit the overexpression of polymerase I and transcript release factor (PTRF).

CONCLUSION: AT-I could alleviate PI-IBS symptoms through downregulation of PTRF and inhibiting the JNK/ iNOS pathway. This study not only provides a scientific basis to clarify the anti-PI-IBS effect of AT-I and its mechanism but also suggests a novel promising therapeutic strategy to treat the PI-IBS.

Key words: atractylenolide Ⅰ, post-infectious irritable bowel syndrome, polymerase Ⅰ and transcript release factor, network pharmacology, MAP kinase signaling system, nitric oxide synthase type Ⅱ

YUAN Jianan, CHENG Kunming, LI Chao, ZHANG Xiang, DING Zeyu, LI Bing, ZHENG Yongqiu. Atractylenolide I ameliorates post-infectious irritable bowel syndrome by inhibiting the polymerase I and transcript release factor and c-Jun N-terminal kinase/inducible nitric oxide synthase pathway[J]. Journal of Traditional Chinese Medicine, 2025, 45(1): 57-65.

Figures/Tables 6

Figure 1 PPI network of 59 Baizhu (Rhizoma Atractylodis Macrocephalae)-IBS common genes and the top 10 core genes A: PPI network of 59 Baizhu (Rhizoma Atractylodis Macrocephalae)-IBS common genes; B: top 10 core genes in the protein-protein interaction network based on the Degree and Betweenness analysis. IBS: irritable bowel syndrome; PPI: protein-protein interaction

Table 1 Potential active components of Baizhu (Rhizoma Atractylodis Macrocephalae)

MOLID	Compound	OB (%)	DL
MOL000022	14-acetyl-12-senecioyl-2E,8Z,10E-atractylentriol	63.37	0.30
MOL000072	8β-ethoxy atractylenolide III	35.95	0.21
MOL000049	3β-acetoxyatractylone	54.07	0.22
MOL000033	(3S,8S,9S,10R,13R,14S,17R)-10,13-dimethyl-17-[(2R,5S)-5-propan-2-yloctan-2-yl]-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol	36.23	0.78
MOL000043	Atractylenolide Ⅰ	37.37	0.15
MOL000044	Atractylenolide Ⅱ	47.5	0.15
MOL000045	Atractylenolide Ⅲ	68.11	0.17
MOL000062	Biatractylolide	17.45	0.81
MOL000057	Diisobutyl phthalate	49.63	0.13
MOL000008	Apigenin	23.06	0.21
MOL000006	Luteolin	36.16	0.25
MOL002902	Ethyl caffeate	103.85	0.07

Figure 2 AT-I ameliorates the symptoms of PI-IBS in rats A: AT-I treatment reduces fecal water content in PI-IBS rats; B: AT-I treatment increases the body weight of PI-IBS rats; C: sucrose consumption in PI-IBS rats increased after AT-I treatment for one day. PI-IBS group: rats are stimulated using a multistimulus pattern consisting of EPSD, TNBS, and CUMS; AT-I (2.5 mg/kg) + PI-IBS:PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 2.5 mg/kg; AT-I (5 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 5 mg/kg; AT-I (10 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 10 mg/kg; AT-I: atractylenolide I; PI-IBS: post-infectious irritable bowel syndrome. The data were presented as the mean ± standard error of mean deviation (n = 6). aP < 0.01, compared with the control group. bP < 0.05, compared with the PI-IBS group.

Table 2 Effects of AT-I on the behavior of rats in OFT

Group	n	Dose (mg·kg^-1·d^-1)	Total distance (cm)	Distance in center (cm)
Control	6	-	18178±1730	992±110^a
PI-IBS	6	-	12125±1203	281±54
PI-IBS+AT-I	6	2.5	14249±1035	531±87^a
PI-IBS+AT-I	6	5	15357±1275	599±72^a
PI-IBS+AT-I	6	10	17410±1317	874±102^a

Figure 3 Representative images of HE staining of the colon sections of PI-IBS rats after AT-I treatment A: Control group; B: PI-IBS group, rats are stimulated using a multistimulus pattern consisting of EPSD, TNBS, and CUMS;C:AT-I (2.5 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 2.5 mg/kg; D: AT-I (5 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 5 mg/kg; E: AT-I (10 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 10 mg/kg. HE: hematoxylin and eosin; PI-IBS: post-infectious irritable bowel syndrome; AT-I: atractylenolide I; EPSD: early postnatal sibling deprivation; TNBS: 2,4,6-trinitrobenzene sulfonic acid; CUMS: chronic unpredictable mild stress; Scale bar = 250 μm.

Figure 4 Western blot analysis of PTRF, P-JNK, JNK, and iNOS expression and relative expression of TNF-α, IL-6, IL-10, and IFN-γ A: Western blot analysis of the expression of PTRF, P-JNK, JNK and iNOS; B: quantitative evaluation of PTRF/GAPDH; C: quantitative evaluation of P-JNK/JNK; D: quantitative evaluation of iNOS/GAPDH; E: relative expression of TNF-α; F: relative expression of IL-6; G: relative expression of IL-10; H: relative expression of IFN-γ. PI-IBS group: rats are stimulated using a multistimulus pattern consisting of EPSD, TNBS, and CUMS; AT-I (2.5 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 2.5 mg/kg; AT-I (5 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 5 mg/kg; AT-I (10 mg/kg) + PI-IBS: PI-IBS rats followed by AT-I treatment intragastrically at a daily dose of 10 mg/kg. PTRF: polymerase I and transcript release factor; P-JNK: phosphorylated JNK; JNK: c-Jun N-terminal kinase; iNOS: inducible nitric oxide synthase; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; TNF-α: tumor necrosis factor α; IL-6: interleukin 6; IL-10: interleukin-10; IFN-γ: interferon-gamma. The data were presented as the mean ± standard error of mean (n = 6). aP < 0.01, compared with the control group; bP < 0.05, compared with the PI-IBS group.

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