目的 通過(guò)網(wǎng)絡(luò)藥理學(xué)及實(shí)驗(yàn)驗(yàn)證探討苦參堿治療急性胰腺炎炎癥反應(yīng)的分子作用機(jī)制。方法 通過(guò)DisGeNET、GeneCards、OMIM數(shù)據(jù)庫(kù)獲取急性胰腺炎相關(guān)靶點(diǎn)，TCMSP數(shù)據(jù)庫(kù)篩選苦參堿作用靶點(diǎn)，取交集得到苦參堿治療急性胰腺炎的潛在靶點(diǎn)。利用DAVID和京都基因與基因百科全書(shū)（KEGG）數(shù)據(jù)庫(kù)進(jìn)行基因本體（GO）功能和KEGG通路富集分析，通過(guò)String數(shù)據(jù)庫(kù)構(gòu)建蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)，結(jié)合cytoHubba算法篩選核心靶點(diǎn)。采用在線(xiàn)分子對(duì)接工具驗(yàn)證苦參堿與核心靶點(diǎn)的結(jié)合能力。通過(guò)雨蛙肽誘導(dǎo)MPC83細(xì)胞損傷模型，CCK-8檢測(cè)細(xì)胞活力，RT-PCR檢測(cè)炎癥因子[白細(xì)胞介素（IL-6）、腫瘤壞死因子-α（TNF-α）、轉(zhuǎn)化生長(zhǎng)因子-β（TGF-β1）、IL-1β]mRNA表達(dá)；通過(guò)雨蛙素、脂肪酶聯(lián)合誘導(dǎo)C57BL/6小鼠急性胰腺炎模型，蘇木精–伊紅（HE）染色觀察胰腺組織病理變化，ELISA檢測(cè)小鼠血清淀粉酶（AMY）、IL-6、TNF-α水平。結(jié)果 網(wǎng)絡(luò)藥理學(xué)分析發(fā)現(xiàn)苦參堿治療急性胰腺炎的潛在靶點(diǎn)共30個(gè)，富集分析顯示，苦參堿治療急性胰腺炎的靶點(diǎn)顯著富集于細(xì)胞因子受體結(jié)合、炎癥反應(yīng)等GO功能，癌癥通路、IL-17信號(hào)通路等KEGG通路。PPI網(wǎng)絡(luò)篩選出前5位核心靶點(diǎn)為IL-6、TNF、TGF-β1、IL-1β、NFκBIA。分子對(duì)接顯示苦參堿與IL-6、TNF、TGFβ1、IL-1β結(jié)合效能穩(wěn)定。細(xì)胞實(shí)驗(yàn)表明，苦參堿可顯著提高損傷細(xì)胞活力（P＜0.05），下調(diào)IL-6、TNF-α、TGF-β1、IL-1β mRNA表達(dá)（P＜0.05）。動(dòng)物實(shí)驗(yàn)顯示，苦參堿高劑量組顯著降低急性胰腺炎小鼠血清AMY、IL-6、TNF-α水平（P＜0.05、0.01），減輕胰腺組織水腫、壞死及炎細(xì)胞浸潤(rùn)。結(jié)論 苦參堿通過(guò)靶向IL-6、TNF、TGF-β1、IL-1β等炎癥相關(guān)基因，抑制炎癥反應(yīng)，改善急性胰腺炎病理?yè)p傷，其作用機(jī)制與調(diào)控細(xì)胞因子信號(hào)通路密切相關(guān)，為急性胰腺炎治療提供了潛在藥物靶點(diǎn)和理論依據(jù)。

Objective To explore the molecular mechanism of matrine in alleviating inflammatory responses in acute pancreatitis by network pharmacology and experimental validation. Methods Acute pancreatitis -related targets were obtained from DisGeNET, GeneCards, and OMIM databases. Matrine’s action targets were screened using the TCMSP database, and the intersection of these targets was identified as potential targets for matrine in treating acute pancreatitis. GO functional and KEGG pathway enrichment analyses were performed using DAVID and KEGG databases. A protein-protein interaction (PPI) network was constructed via the String database, and core targets were screened using the cytoHubba algorithm. Online molecular docking tools were used to validate the binding ability of matrine to core targets. A cerulein-induced MPC83 cell injury model and cell viability were detected by CCK-8 assay. The expression of inflammatory cytokines (IL-6, TNF-α, TGF-β1, IL-1β) mRNA was detected by RT-PCR. A cerulein/lipase-induced C57BL/6 mice acute pancreatitis model was used. Pathological changes in pancreatic tissues were observed via HE staining. Serum levels of AMY, IL-6, and TNF-α in mice were measured by ELISA. Results A total of 30 potential targets for matrine in acute pancreatitis treatment were identified. Enrichment analysis showed that these targets were significantly enriched in GO such as cytokine receptor binding, inflammatory response, as well as KEGG pathways such as pathways in cancer, IL-17 signaling pathway. The top 5 core targets screened from the PPI network were IL-6, TNF, TGFβ1, IL-1β, and NFκBIA. Molecular docking showed that matrine had stable binding efficacy with IL-6, TNF, TGFβ1 and IL-1β. Cell experiments showed that matrine significantly enhanced the viability of injured cells (P< 0.05), and downregulated the mRNA expression of IL-6, TNF-α, TGF-β1, and IL-1β (P< 0.05). Animal experiments showed that matrine group significantly reduced serum AMY, IL-6, and TNF-α levels (P< 0.05, 0.01) in acute pancreatitis mice, and alleviated pancreatic tissue edema, necrosis, and inflammatory cell infiltration. Conclusion Matrine inhibits the inflammatory response and improves acute pancreatitis pathological damage by targeting inflammation-related genes such as IL-6, TNF, TGF-β1, and IL-1β. Its mechanism is closely related to regulating cytokine signaling pathways, providing potential drug targets and a theoretical basis for acute pancreatitis treatment.

R285.5

泰安市科技發(fā)展計(jì)劃（引導(dǎo)計(jì)劃）項(xiàng)目（2021NS421，2022NS152）