摘要
目的 探讨小檗碱对葡聚糖硫酸钠(DSS)诱导的Balb/c小鼠溃疡性结肠炎模型的保护作用,并采用蛋白质组学技术分析其分子作用机制。方法 将24只Balb/c雄性小鼠随机分为对照组、溃疡性结肠炎模型组及小檗碱干预组,每组8只,通过饮用3%DSS溶液建立溃疡性结肠炎模型,小檗碱干预组每日灌胃40mg/kg小檗碱,干预7d后处理各组小鼠。采用疾病活动指数(DAI)评估结肠炎严重程度,检测血清炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-1β]与氧化应激指标[超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)],结合异硫氰酸荧光素-葡聚糖检测结肠通透性评估小檗碱疗效。采用蛋白质组学技术筛选结肠组织差异蛋白,并采用RT-qPCR和蛋白质印迹法验证关键基因及蛋白表达。结果 小檗碱显著缓解了DSS诱导的溃疡性结肠炎小鼠的病理损伤,与溃疡性结肠炎模型组相比,小檗碱干预组小鼠的体重下降趋势明显改善,DAI评分显著降低。小檗碱有效抑制了血清TNF-α、IL-6和IL-1β的上升,并逆转了SOD、MDA及CAT的活性。此外,小檗碱通过上调紧密连接蛋白Occludin和Zo-1的表达,显著修复了肠上皮屏障功能,且降低了肠道通透性。蛋白质组学分析显示,小檗碱干预后结肠组织中鉴定出142个差异表达蛋白(77个上调,65个下调),并显著激活过氧化物酶体增殖物激活受体γ(PPARγ)信号通路。RT-qPCR和蛋白质印迹实验表明,小檗碱可通过上调肉碱棕榈酰转移酶1、脂肪酸转运蛋白2、PPARγmRNA和蛋白的表达调控代谢-炎症网络,从而治疗溃疡性结肠炎。结论 小檗碱通过抑制系统性炎症反应、缓解氧化应激损伤、修复肠上皮屏障完整性,并激活PPARγ信号通路,多靶点治疗DSS诱导的小鼠溃疡性结肠炎。
Objective To investigate the protective effect of berberine on dextran sulphate sodium salt(DSS)-induced ulcerative colitis(UC)in Balb/c mice and to analyze its molecular mechanism using proteomics technology.Methods Twenty-four male Balb/c mice were randomly divided into a control group,a UC model group,and a berberine intervention group.The UC model was established by administering a 3%DSS solution,and the berberine intervention group received daily oral administration of 40 mg/kg berberine.Mice in each group were treated after intervention for 7 d.The disease activity index(DAI)was used to assess the severity of colitis.Serum inflammatory factors,including tumour necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β,as well as oxidative stress markers,including superoxide dismutase(SOD),malondialdehyde(MDA),and catalase(CAT)were detected.Colon permeability was assessed using fluorescein isothiocyanate-dextran to evaluate the efficacy of berberine.Proteomics technology was used to screen for differentially expressed proteins in colon tissue,and RT-qPCR and Western blotting were employed to validate the expression of key genes and proteins.Results Berberine significantly alleviated pathological damage in DSS-induced UC mice.Compared with the UC model group,the weight loss trend in the berberine intervention group was significantly improved,and the DAI score was significantly reduced.Berberine effectively inhibited the elevation of serum pro-inflammatory factors TNF-α,IL-6,and IL-1β,and reversed the activity of SOD,MDA,and CAT.Additionally,berberine significantly repaired intestinal epithelial barrier function by upregulating the expression of tight junction proteins Occludin and Zo-1,and reduced intestinal permeability.Proteomics analysis revealed that 142 differentially expressed proteins(77 up-regulated and 65 down-regulated)were identified in colon tissue after berberine intervention,and the peroxisome proliferator-activated receptorγ(PPARγ)signaling pathway was significantly activated.RT-qPCR and Western blot experiments indicated that berberine could regulate the metabolism-inflammation network by upregulating the expressions of CPT1,FATP2,PPARγmRNA,and protein,thereby improving UC.Conclusion Berberine improves DSS-induced UC in mice through multiple mechanisms,including the inhibition of systemic inflammatory responses,alleviation of oxidative stress damage,repair of intestinal epithelial barrier integrity,and activation of the PPARγsignaling pathway.
作者
华宏军
徐可珍
杨超
张芸芸
王晓波
叶旭星
HUA Hongjun;XU Kezhen;YANG Chao;ZHANG Yunyun;WANG Xiaobo;YE Xuxing(Department of Gastroenterology,Jinhua Municipal Central Hospital,Jinhua 321000,China;不详)
出处
《浙江医学》
2025年第17期1799-1805,I0001,共8页
Zhejiang Medical Journal
基金
浙江省基础公益研究计划项目(LGD19H030002)。
作者简介
通信作者:叶旭星,E-mail:clive4343@163.com。
