Mechanistic study of the therapeutic effect of Portulaca oleracea L on colorectal inflammatory cancer transformation in mice by inhibiting Caspase 12 protein based on proteomics
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摘要: 目的 探讨马齿苋通过Caspase 12介导的NOD信号通路对结直肠炎癌转化小鼠的保护作用机制。方法 将24只6~8周龄的C57小鼠，随机分为正常对照组、模型组、马齿苋治疗组。模型组和马齿苋治疗组采用氧化偶氮甲烷(AOM)联合葡聚糖硫酸钠(DSS)诱导结直肠炎相关肿瘤发生小鼠模型，马齿苋治疗组每日按照人鼠等效剂量灌胃；通过各组小鼠的脾体比、结直肠长度、肠道息肉/肿瘤总数及肠道组织的苏木精-伊红染色，分析马齿苋抑制小鼠结直肠炎癌转化的药效；通过蛋白组分析、Western blot检测，验证马齿苋对NOD信号通路关键蛋白Caspase 12的调控；分别通过qPCR法和免疫荧光法检测小鼠组织中Caspase 12调控的细胞因子IL-1β、趋化因子和巨噬细胞标志物F4/80的表达；最后采用Alican blue和PAS病理染色检测肠道黏蛋白的水平，使用Western blot检测肠道紧密连接蛋白的水平，以评价马齿苋对肠道屏障的保护作用。结果 马齿苋明显抑制小鼠结直肠炎癌转化进展，减少肠道息肉/肿瘤数量和肠道Ki67水平，马齿苋可显著改善结直肠炎癌转化小鼠NOD样受体信号通路功能，抑制关键蛋白Caspase 12的水平，减少下游细胞因子IL-1β和趋化因子MCP-1含量，减少结直肠炎癌转化小鼠中肠道组织巨噬细胞的表达，改善结直肠炎癌转化小鼠肠道屏障功能。结论 马齿苋能够抑制小鼠结直肠炎癌转化进展，抑制炎症反应，改善肠道屏障，其作用可能与调控Caspase 12介导的NOD样受体信号通路有关。
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关键词:
- 结直肠炎癌转化 /
- 马齿苋 /
- Caspase 12 /
- 肠道屏障
Abstract: Objective To investigate the mechanism of the protective effect of Portulaca oleracea L on colorectal inflammatory cancer transformation model mice through the NOD signaling pathway mediated by Caspase 12.Methods Twenty-four C57 mice aged 6-8 W were randomly divided into normal control, model control, and portulaca oleracea L treatment groups. In the model control group and portulaca oleracea L treatment group, Azoxymethane (AOM) combined with Dextran sodium sulfate (DSS) was used to induce colitis-associated tumorigenesis mice model, and in the portulaca oleracea L treatment group, gavage the drug every day according to the equivalent dose of human mice. The efficacy of Portulaca oleracea L in inhibiting colorectal cancer transformation mice was analyzed by ratio of spleen/body, colorectal length, total number of polyps/tumors, and HE staining of intestinal tissues in each group. Proteomic analysis and Western blot were used to clarify the therapeutic effect of the NOD signaling pathway, especially the key protein Caspase 12. The expression of cytokine IL-1β and macrophage marker F4/80 in mice tissues were detected by qPCR and immunofluorescence, respectively. Finally, the level of intestinal mucin was detected by Alican blue and PAS pathological staining, and the level of intestinal tight junction protein was detected by Western blot, to evaluate the protective effect of Portulaca oleracea L on the intestinal barrier.Results Portulaca oleracea L significantly inhibited the progression of colorectal inflammatory cancer transformation in mice, and reduced the number of intestinal polyps/tumors and intestinal Ki67 levels. Portulaca oleracea L significantly improved the function of the NOD-like receptor signaling pathway in colorectal inflammatory cancer transformation mice, inhibited the level of the key protein Caspase 12, reduced the levels of the downstream cytokine IL-1β and chemokine MCP-1, and inhibited the expression of macrophages in the intestinal tissues. Portulaca oleracea L improved the intestinal barrier function in colorectal inflammation cancer-transformed mice.Conclusion Portulaca oleracea L significantly inhibited the progression of colorectal inflammatory cancer transformation in mice models, suppressed inflammatory responses, and improved the intestinal barrier, and its effects may be related to the modulation of the Caspase 12-mediated NOD-like receptor signaling pathway. -
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