摘要
目的:本研究旨在探讨美沙拉嗪(MSLZ)对小鼠2,4,6-三硝基苯磺酸(TNBS)模型中Let-7i-5p和TLR4/MyD88依赖通路的调控机制。方法:采用TNBS/乙醇结肠法建立溃疡性结肠炎(UC)模型。将44只雄性小鼠随机分为对照组、模型组、MSLZ组和Let-7i-5p抑制剂组,每组11只。小鼠灌胃或腹腔注射相应的药物或生理盐水,连续14d。采用实时荧光定量聚合酶链反应(qRT-PCR)检测小鼠结肠组织中和血清中Let-7i-5p的表达水平。在显微镜下,用苏木精和伊红(HE)染色观察结肠组织的病理病变。分别采用qRT-PCR,Western blot和免疫组化方法检测小鼠结肠组织中TLR4/MyD88依赖通路相关基因的mRNA和蛋白水平。通过ELISA检测小鼠血清中TNF-α和IL-1β的表达水平。结果:根据疾病活跃指数(DAI)、结肠损伤和病理病变的评分,成功构建了小鼠UC模型。模型组小鼠结肠组织中和血清中Let-7i-5p的表达水平显著高于对照组(P<0.0001)。与模型组相比,MSLZ和Let-7i-5p抑制剂处理均能显著抑制Let-7i-5p的表达(P<0.0001)。与对照组相比,模型组小鼠结肠组织中TLR4/MyD88依赖通路相关基因(包括TLR4、MyD88、TRAF-6和NF-κB)的mRNA和蛋白水平显著上调。MSLZ和Let-7i-5p抑制剂处理均能显著抑制这些基因的表达,且MSLZ的抑制作用略强于Let-7i-5p抑制剂。与对照组相比,模型组小鼠结肠组织中IL-1β和TNF-α的mRNA水平和血清中的蛋白水平显著上调,MSLZ和Let-7i-5p抑制剂处理均能抑制IL-1β和TNF-α的表达水平。结论:在TNBS/乙醇诱导的UC小鼠模型中,MSLZ可以抑制结肠组织中和血清中Let-7i-5p的表达。此外,MSLZ还通过抑制UC小鼠的TLR4/MyD88依赖性通路来抑制炎症因子的释放。
Objective:To investigate the regulatory mechanism of mesalazine(MSLZ)on Let-7i-5p and TLR4/MyD88 signaling pathway in a mouse model of 2,4,6-trinitrobenzene sulfonic acid(TNBS)-in-duced ulcerative colitis(UC).Methods:A UC model was established using the TNBS/ethanol method.Forty-four male mice were randomly divided into four groups:control group,model group,MSLZ group,and Let-7i-5p inhibitor group,with 11 mice in each group.Mice were gavaged or intraperitoneally injected with corre-sponding drugs or saline for 14 consecutive days.The expression levels of Let-7i-5p in colon tissues and ser-um of mice were detected by real-time quantitative polymerase chain reaction(qRT-PCR).The pathological changes of colon tissues were observed by hematoxylin and eosin(HE)staining under a microscope.The mR-NA and protein levels of TLR4/MyD88 signaling pathway-related genes in colon tissues of mice were detected by qRT-PCR,Western blot,and immunohistochemistry,respectively.The expression levels of TNF-αand IL-1βin mouse serum were detected by ELISA.Results:According to the disease activity index(DAI),co-lon damage score,and pathological lesion score,the mouse UC model was successfully established.The ex-pression levels of Let-7i-5p in colon tissues and serum of mice in the model group were significantly higher than those in the control group(P<0.0001).Compared with the model group,both MSLZ and Let-7i-5p in-hibitor treatment could significantly inhibit the expression of Let-7i-5p(P<0.0001).Compared with the control group,the mRNA and protein levels of TLR4/MyD88 signaling pathway-related genes(including TLR4,MyD88,TRAF-6,and NF-κB)in colon tissues of mice in the model group were significantly upregu-lated.MSLZ and Let-7i-5p inhibitor treatment could significantly inhibit the expression of these genes,and the inhibitory effect of MSLZ was slightly stronger than that of Let-7i-5p inhibitor.Compared with the control group,the mRNA levels of IL-1βand TNF-αin colon tissues and the protein levels in serum of mice in the model group
作者
毛珍珍
刘靖
张晨华
闫娜娜
赵玲玲
卢军仪
许伟光
王婧
MAO Zhenzhen;LIU Jing;ZHANG Chenhua(Cangzhou Central Hospital of Traditional Chinese and Western Medicine/Cangzhou Second People's Hospital,Hebei Cangzhou 061000,China)
出处
《河北医学》
CAS
2024年第4期549-555,共7页
Hebei Medicine
基金
河北省中医药管理局科研计划项目(编号:2024178)。
