南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (7): 1685-1695.doi: 10.12122/j.issn.1673-4254.2026.07.21
• • 上一篇
张龙涛1,3(
), 章雨1, 乔通3, 张可妮1, 尹林1, 黄菊2, 李静1,2, 耿志军2, 胡建国1,2(
)
收稿日期:2026-01-30
出版日期:2026-07-20
发布日期:2026-07-20
通讯作者:
胡建国
E-mail:15055561902@163.com;jghu9200@bbmu.edu.cn
作者简介:张龙涛,在读硕士研究生,E-mail:15055561902@163.com
基金资助:
Longtao ZHANG1,3(
), Yu ZHANG1, Tong QIAO3, Keni ZHANG1, Lin YIN1, Ju HUANG2, Jing LI1,2, Zhijun GENG2, Jianguo HU1,2(
)
Received:2026-01-30
Online:2026-07-20
Published:2026-07-20
Contact:
Jianguo HU
E-mail:15055561902@163.com;jghu9200@bbmu.edu.cn
摘要:
目的 探讨丁香酸甲酯(MS)对葡聚糖硫酸钠(DSS)诱导小鼠结肠炎的保护作用及机制。 方法 将24只C57BL/6小鼠随机分为对照组(Con组)、造模组(DSS组)、药物处理组(MS组,100 mg/kg),8只/组。通过检测小鼠体质量、疾病活动度(DAI)评分、结肠长度、HE与AB-PAS染色及组织学评分,评估MS对结肠炎的治疗效果。采用ELISA和RT-qPCR检测结肠炎症因子IL-6、TNF-α和IL-10的表达,免疫组化检测髓过氧化物酶(MPO)在结肠组织中的表达,免疫荧光和Western blotting检测紧密连接蛋白ZO-1、Claudin-1的表达与分布,TUNEL染色检测结肠凋亡细胞。体外采用1% DSS诱导NCM460细胞构建凋亡模型,给予MS(50 μmol/L)干预后,通过流式细胞术检测细胞凋亡。采用网络药理学预测和Western blotting检测分析MS的作用机制。 结果 MS处理改善了DSS引起的小鼠体质量下降、结肠缩短、DAI评分和组织学评分升高,减轻肠绒毛结构损伤,增加杯状细胞数量(P<0.05)。同时MS可下调小鼠肠黏膜组织中IL-6、TNF-α和MPO的表达,并上调IL-10的表达(P<0.05)。免疫荧光与Western blotting表明MS可恢复紧密连接蛋白ZO-1、Claudin-1的表达与分布。TUNEL、流式细胞术及Western blotting结果一致表明,MS在体内外均能显著降低肠上皮细胞的凋亡比例,上调抗凋亡蛋白Bcl-2和XIAP,下调促凋亡蛋白C-Caspase3(P<0.05)。KEGG富集分析提示MAPK通路可能与MS疗效相关。Western blotting进一步证实MS能抑制体内外模型中p-JNK、p-ERK、p-p38的磷酸化水平(P<0.05)。 结论 MS通过减少肠上皮细胞凋亡和改善肠屏障损伤来缓解DSS诱导的小鼠结肠炎,其机制可能与抑制MAPK信号通路的表达有关。
张龙涛, 章雨, 乔通, 张可妮, 尹林, 黄菊, 李静, 耿志军, 胡建国. 丁香酸甲酯通过抑制MAPK通路调控肠上皮细胞凋亡减轻葡聚糖硫酸钠诱导的小鼠结肠炎[J]. 南方医科大学学报, 2026, 46(7): 1685-1695.
Longtao ZHANG, Yu ZHANG, Tong QIAO, Keni ZHANG, Lin YIN, Ju HUANG, Jing LI, Zhijun GENG, Jianguo HU. Methyl syringate alleviates DSS-induced colitis in mice by suppressing intestinal epithelial cell apoptosis via inhibiting the MAPK signaling pathway[J]. Journal of Southern Medical University, 2026, 46(7): 1685-1695.
图1 MS处理对DSS诱导的小鼠结肠炎疾病状态的影响
Fig.1 Effect of methyl syringate (MS) treatment on disease status in dextran sodium sulfate (DSS)-induced colitis in mice. A, B: Changes in body weight and DAI scores of the mice. C, D: Comparison of colon lengths among the groups. Data are presented as Mean±SD (n=8). *P<0.05 vs Con group; #P<0.05 vs DSS group.
图2 MS处理对DSS诱导的小鼠结肠组织的影响
Fig.2 Effect of MS treatment on colonic pathology in mice with DSS-induced colitis. A: HE staining of colon tissue in different groups. B: Changes in histological scores (Mean±SD, n=8). C: AB-PAS staining of colon tissue in different groups. D: Goblet cell count (Mean±SD, n=3). *P<0.05 vs Con group; #P<0.05 vs DSS group.
图3 MS处理对DSS模型小鼠肠道炎症因子水平的影响
Fig.3 Effect of MS treatment on intestinal inflammatory cytokines in the colon tissues of mice with DSS-induced colitis. A: ELISA results of IL-6, TNF-α and IL-10 in the intestinal mucosa of the mice. B: RT-qPCR results of IL-6, TNF-α and IL-10 in the intestinal mucosa of the mice. C: MPO immunohistochemical staining results. D: Relative integrated optical density value of MPO. Data are presented as Mean±SD (n=3). *P<0.05 vs Con group; #P<0.05 vs DSS group.
图4 MS处理对DSS模型小鼠结肠炎肠屏障的影响
Fig.4 Effect of MS treatment on intestinal barrier function of the mice with DSS-induced colitis. A-C: 3D immunofluorescence staining and the mean fluorescence intensities for ZO-1 and claudin-1. D: Protein levels of ZO-1 and claudin-1 detected by Western blotting. E: Relative protein expression levels of ZO-1 and claudin-1. Data are presented as Mean±SD (n=3). *P<0.05 vs Con group. #P<0.05 vs DSS group.
图5 网络药理学分析结果
Fig.5 Results of network pharmacology analysis. A: Venn diagram of MS and IBD. B: PPI network of intersecting targets between MS and IBD. C: Bar chart of GO enrichment analysis for MS and IBD. D: Bubble plot of KEGG pathway enrichment analysis for MS and IBD.
图7 MS处理对DSS模型小鼠结肠上皮细胞凋亡的影响
Fig.7 Effect of MS treatment on apoptosis of colon epithelial cells in DSS-treated mice. A: TUNEL staining for detecting apoptosis in intestinal epithelial cells. B: Apoptosis cell ratio of intestinal epithelial cells. C: Protein expression levels of XIAP, Bcl-2, and cleaved caspase-3 in mouse colon tissues detected by Western blotting. D: Relative protein expression levels of XIAP, Bcl-2, and cleaved caspase-3 in mouse colon tissue. Data are presented as Mean±SD (n=3). *P<0.05 vs Con group; #P<0.05 vs DSS group.
图8 MS处理对DSS诱导的NCM460细胞凋亡的影响
Fig.8 Effect of MS treatment on DSS-induced apoptosis in NCM460 cells. A: Apoptosis of NCM460 cells detected by flow cytometry. B: Apoptosis cell ratio of NCM460 cells. C: Protein expression levels of XIAP, Bcl-2, and cleaved caspase-3 in NCM460 cells detected by Western blotting. D: Relative protein expression levels of XIAP, Bcl-2, and cleaved caspase-3 in NCM460 cells. Data are presented as Mean±SD (n=3). *P<0.05 vs Con group; #P<0.05 vs DSS group.
图9 MS处理对MAPK通路活化的影响
Fig.9 Effect of MS treatment on MAPK pathway activation. A: Protein expression levels of p-ERK, ERK, p-JNK, JNK, p-p38, and p38 in mouse colon mucosal tissues detected by Western blotting. B: Relative protein expression levels of p-ERK, p-JNK, and p-p38 in mouse colon mucosal tissues. C: Protein expression levels of p-ERK, ERK, p-JNK, JNK, p-p38, and p38 in NCM460 cells detected by Western blotting. D: Relative protein expression levels of p-ERK, p-JNK, and p-p38 in NCM460 cells. Data are presented as Mean±SD (n=3). *P<0.05 vs Con group; #P<0.05 vs DSS group.
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