YTHDC1介导的XBP1 m6A修饰在解毒化瘀汤促进自噬治疗溃疡性结肠炎中的机制研究OA
Study on the Mechanism of Jiedu Huayu Decoction Promoting Autophagy in the Treatment of Ulcerative Colitis Based on YTHDC1-Mediated XBP1 m6A Modification
目的 观察YTHDC1介导的XBP1 mRNA m6A甲基化修饰在解毒化瘀汤调控细胞自噬治疗溃疡性结肠炎(UC)中的作用机制.方法 将60只SD大鼠随机分为对照组、模型组、解毒化瘀汤低、中、高剂量组及阳性药物组.采用5％葡聚糖硫酸钠(DSS)自由饮水7 d建立UC模型,同时灌胃给药干预7 d.采用苏木精-伊红(HE)染色观察结肠组织病理变化;酶联免疫吸附分析(ELISA)检测血清炎症因子水平;透射电镜观察结肠组织超微结构;蛋白质印迹法(Western blotting)检测自噬相关蛋白、YTHDC1及XBP1s蛋白的表达;定量逆转录聚合酶链反应(qRT-PCR)检测YTHDC1和XBP1 mRNA表达;MeRIP-qPCR检测XBP1 mRNA的m6A修饰水平.结果 与模型组相比,解毒化瘀汤各剂量组能显著改善DSS诱导的结肠组织损伤,降低炎症评分及血清促炎因子水平(P＜0.01).同时,解毒化瘀汤呈剂量依赖性增加结肠上皮细胞中自噬体数量,上调LC3-Ⅱ/Ⅰ及p-AMPK/AMPK的比值,下调p62的表达及p-mTOR/mTOR的比值(P＜0.01).此外,解毒化瘀汤显著上调了模型大鼠结肠组织中YTHDC1的蛋白及mRNA表达、XBP1s蛋白及XBP1 mRNA的表达(P＜0.01),并逆转了 DSS导致的XBP1 mRNA m6A修饰水平的下调,差异均有统计学意义(P＜0.01).结论 解毒化瘀汤可能通过上调YTHDC1表达,增加XBP1 mRNA的m6A修饰,进而增加XBP1s蛋白的水平,激活AMPK/mTOR通路介导的自噬,抑制肠道炎症,发挥治疗UC的作用.
Objective:To explore the mechanism of Jiedu Huayu Decoction regulating autophagy in the treat-ment of ulcerative colitis(UC)based on YTHDC1-mediated XBP1 mRNA m6A methylation modification.Meth-ods:Sixty SD rats were randomly divided into the control group,model group,low-,medium-and high-dose Jie-du Huayu Decoction groups,and positive drug group.UC model was established by allowing free access to 5％dex-tran sulfate sodium(DSS)for 7 days,while intragastric administration was given simultaneously during the same period.Pathological changes of colon tissue were observed by HE staining;serum inflammatory factor levels were detected by enzyme-linked immunosorbent assay(ELISA);ultrastructure of colon tissue was observed by transmis-sion electron microscope;the expression of autophagy-related proteins,YTHDC1 and XBP1s was detected by West-ern blotting;the mRNA expression of YTHDC1 and XBP1 was detected by quantitative reverse transcription-poly-merase chain reaction(qRT-PCR);the m6A modification level of XBP1 mRNA was detected by MeRIP-qPCR.Results:Compared with the model group,all dose groups of Jiedu Huayu Decoction significantly improved DSS-in-duced colon tissue injury,and reduced inflammatory score and serum pro-inflammatory factor levels(P＜0.01).Meanwhile,Jiedu Huayu Decoction dose-dependently increased the number of autophagosomes in colonic epitheli-al cells,up-regulated the ratios of LC3-Ⅱ/Ⅰ and p-AMPK/AMPK,and down-regulated the expression of p62 and the ratio of p-mTOR/mTOR(P＜0.01).In addition,Jiedu Huayu Decoction significantly up-regulated the pro-tein and mRNA expression of YTHDC1,the protein expression of XBP1s and mRNA expression of XBP1 in colon tissue of model rats(P＜0.01),and reversed the down-regulation of XBP1 mRNA m6A modification level induced by DSS,with statistically significant differences(P＜0.01).Conclusion:Jiedu Huayu Decoction may up-regulate YTHDC1 expression,increase m6A modification of XBP1 mRNA,thereby elevate the level of XBP1s protein,acti-vate AMPK/mTOR pathway-mediated autophagy,inhibit intestinal inflammation,and exert therapeutic effect on UC.
刘颖;王鹏;袁星星;李丽萍
黑龙江中医药大学附属第一医院,黑龙江哈尔滨 150040黑龙江中医药大学附属第一医院,黑龙江哈尔滨 150040黑龙江省中医药科学院,黑龙江哈尔滨 150006黑龙江省中医药科学院,黑龙江哈尔滨 150006
医药卫生
溃疡性结肠炎解毒化瘀汤YTHDC1m6A甲基化自噬大鼠
Ulcerative colitisJiedu Huayu DecoctionYTHDC1m6A methylationAutophagyRats
《中国中医急症》 2026 (5)
521-527,7
中华中医药学会青年培英计划项目(202557-011)北京医卫健康公益基金会医学科学研究基金项目(YWJKJJHKYJJ-B2025032601)
评论