The active ingredient β-sitosterol in the anti-inflammatory agents alleviates perianal inflammation in rats by inhibiting the expression of Srebf2, activating the PPAR signaling pathway, and altering the composition of gut microbiota

IF 4.7 2区医学 Q2 IMMUNOLOGY International immunopharmacology Pub Date : 2025-04-16 Epub Date: 2025-03-13 DOI:10.1016/j.intimp.2025.114470

Yanlan Wu , Hao Ge , Haoran Zhao , Kaiping Zou , Pei Wang , Yi Wang , Yang Zhang

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Abstract

Background

Anti-inflammatory herbal formulations are common in traditional Chinese medicine for clearing heat and detoxifying; however, the specific active components and their mechanisms remain unclear.

Objective

This study investigates the role of Sitosterol in alleviating perianal inflammation and its underlying mechanisms.

Methods

Sitosterol was identified as a key active ingredient through the TCMSP database. Its structure was analyzed using PubChem, target genes were explored with STITCH, and KEGG pathways related to Srebf2 were revealed by STRING. An animal model of perianal inflammation was induced with 75 % acetic acid and treated with Sitosterol, water, normal saline, or antibiotics. The effects on gut microbiota were assessed using 16S rRNA sequencing, and inflammation was evaluated through HE stains, IHC, and TUNEL assays. In vitro, LPS-treated Caco-2 cells were used to measure proliferation, apoptosis, and cytokine levels, with PPAR pathway involvement examined using GW6471.

Results

Sitosterol emerged as the primary active ingredient targeting Srebf2, with KEGG analysis highlighting the PPAR signaling pathway. In rats, Sitosterol reduced weight loss, inflammatory cell infiltration, edema, and vasodilation in perianal tissue. Additionally, it decreased PCNA levels, increased apoptosis, and elevated serum levels of IL-1β, IL-6, and TNF-α, particularly at high doses compared to antibiotics. Sitosterol also restored gut microbiota. Srebf2 knockdown improved tissue conditions and modulated cytokine levels, effects that were countered by GW6471. In LPS-treated Caco-2 cells, Sitosterol reversed reductions in cell viability and proliferation and modulated the expression of proteins and cytokines.

Conclusion

Sitosterol restores gut microbiota composition and further alleviates perianal inflammation in rats by inhibiting Srebf2 expression and activating the PPAR signaling pathway.

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抗炎剂中的有效成分β-谷甾醇通过抑制 Srebf2 的表达、激活 PPAR 信号通路和改变肠道微生物群的组成，缓解大鼠的肛周炎症

抗炎中药制剂在中药中用于清热解毒是很常见的；然而，具体的有效成分及其机制尚不清楚。目的探讨谷甾醇在缓解肛周炎症中的作用及其机制。方法通过TCMSP数据库鉴定谷甾醇为其关键活性成分。利用PubChem分析其结构，利用STITCH探索靶基因，利用STRING揭示与Srebf2相关的KEGG通路。采用75%醋酸致肛周炎症动物模型，谷甾醇、水、生理盐水或抗生素处理。使用16S rRNA测序评估对肠道微生物群的影响，并通过HE染色，IHC和TUNEL检测评估炎症。体外，使用lps处理的Caco-2细胞测量增殖、凋亡和细胞因子水平，并使用GW6471检测PPAR通路的参与情况。结果谷甾醇是靶向Srebf2的主要活性成分，KEGG分析突出了PPAR信号通路。在大鼠中，谷甾醇减轻了体重减轻、炎症细胞浸润、水肿和肛周组织血管舒张。此外，与抗生素相比，它降低了PCNA水平，增加了细胞凋亡，升高了血清IL-1β、IL-6和TNF-α水平，特别是在高剂量时。谷甾醇还能恢复肠道微生物群。Srebf2敲除改善了组织状况并调节了细胞因子水平，而GW6471则抵消了这一作用。在lps处理的Caco-2细胞中，谷甾醇逆转了细胞活力和增殖的降低，并调节了蛋白质和细胞因子的表达。结论谷甾醇通过抑制Srebf2的表达和激活PPAR信号通路，恢复大鼠肠道菌群组成，进一步缓解肛周炎症。

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来源期刊

International immunopharmacology 医学-免疫学

CiteScore

8.40

自引率

3.60%

发文量

935

审稿时长

53 days

期刊介绍： International Immunopharmacology is the primary vehicle for the publication of original research papers pertinent to the overlapping areas of immunology, pharmacology, cytokine biology, immunotherapy, immunopathology and immunotoxicology. Review articles that encompass these subjects are also welcome. The subject material appropriate for submission includes: • Clinical studies employing immunotherapy of any type including the use of: bacterial and chemical agents; thymic hormones, interferon, lymphokines, etc., in transplantation and diseases such as cancer, immunodeficiency, chronic infection and allergic, inflammatory or autoimmune disorders. • Studies on the mechanisms of action of these agents for specific parameters of immune competence as well as the overall clinical state. • Pre-clinical animal studies and in vitro studies on mechanisms of action with immunopotentiators, immunomodulators, immunoadjuvants and other pharmacological agents active on cells participating in immune or allergic responses. • Pharmacological compounds, microbial products and toxicological agents that affect the lymphoid system, and their mechanisms of action. • Agents that activate genes or modify transcription and translation within the immune response. • Substances activated, generated, or released through immunologic or related pathways that are pharmacologically active. • Production, function and regulation of cytokines and their receptors. • Classical pharmacological studies on the effects of chemokines and bioactive factors released during immunological reactions.