| 陈 梦,马红梅,曾文莉,方 婷,阮 雷.基于TLR4MyD88信号通路观察粪菌移植对DSS诱导小鼠溃疡性结肠炎干预作用机制[J].现代生物医学进展英文版,2020,(24):4606-4611. |
| 基于TLR4MyD88信号通路观察粪菌移植对DSS诱导小鼠溃疡性结肠炎干预作用机制 |
| Based on the TLR4/MyD88 Signaling Pathway, the Mechanism of Fecal Bacteria Transplantation on the Intervention of Dss-induced Ulcerative Colitis in Mice was Observed |
| Received:April 29, 2020 Revised:May 24, 2020 |
| DOI：10.13241/j.cnki.pmb.2020.24.002 |
| 中文关键词: 溃疡性结肠炎 TLR4/MyD88信号通路 NF-κB 细胞因子 |
| 英文关键词: Ulcerative colitis TLR4/MyD88 signaling pathway NF-κB Cytokines |
| 基金项目:湖北省自然科学基金项目(2015CKB754) |
|
| Hits: 755 |
| Download times: 547 |
| 中文摘要: |
| 摘要 目的：探讨粪菌移植（fecal microbiota transplantation，FMT）对葡聚糖硫酸钠（dextran sodium sulfate, DSS）诱导小鼠溃疡性结肠炎（ulcerative colitis，UC）干预作用及其炎症相关的分子机制。方法：30只昆明小鼠采用DSS喂养建立结肠炎小鼠模型并随机分为模型组、FMT低剂量组和FMT高剂量组，另取10只作为对照组。FMT低剂量组、FMT高剂量组自造模后第1 d开始分别给予8 g/kg、15 g/kg粪菌剂量灌肠处理，灌肠体积0.2 mL。对照组和模型组给予等体积生理盐水灌肠处理。分别在实验第1、7、14及21 d称量小鼠体重。实验结束后处死小鼠，取出结直肠，观察各组小鼠结直肠形态变化并通过HE染色观察病变程度。ELISA检测小鼠结直肠组织匀浆液上清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin 6, IL-6）、白细胞介素-4（interleukin 4, IL-4）和白细胞介素-10（interleukin 10, IL-10）的表达变化；免疫组化法和RT-qPCR分别检测结直肠组织中核因子κB-P65亚基 (nuclear factor-κB P65, NF-κB P65)蛋白和mRNA的表达情况；Western blotting检测结直肠组织中Toll 样受体4 (Toll-like receptor 4，TLR4)、髓样分化因子 88(myeloid differentiation factor 88，MyD88)的蛋白表达情况。结果：对照组相比，造模小鼠在第6 d起至第14 d体重明显减轻，差异具有统计学意义（P<0.05）；与模型组相比，FMT低、高剂量组小鼠第10 d起至第14 d体重明显减轻，FMT高剂量组小鼠体重升高更加明显，差异具有统计学意义（P<0.05）；与对照组相比，模型组小鼠结直肠组织中TNF-α、IL-6、NF-κB P65 mRNA和蛋白阳性表达、TLR4、MyD88 蛋白表达均显著升高(P<0.05)，IL-4、IL-10表达显著下降（P<0.05）；FMT高剂量组IL-4、IL-10较模型组显著升高，其余指标均显著下降（P<0.05）。结论：FMT可通过抑制TLR4/MyD88/NF-κB信号通路缓解炎症反应，发挥对UC的治疗作用。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of fecal microbiota transplantation (FMT) on mice ulcerative colitis (UC) induced by dextran sodium sulfate (DSS) and the molecular mechanism of inflammation. To investigate the effect of fecal microbiota transplantation (FMT) on mice ulcerative colitis (UC) induced by dextran sodium sulfate (DSS) and the molecular mechanism of inflammation. Methods: Thirty kunming mice were fed DSS to establish a model of colitis and randomly divided into the model group, the low-dose FMT group and the high-dose FMT group. The low-dose FMT group and the high-dose FMT group were given 8 g/kg and 15 g/kg fecal bacteria dose enema, with the enema volume of 0.2 mL, from day 1 after modeling. The control group and the model group were given equal volume normal saline enema. The mice were weighed on day 1, 7, 14 and 21, respectively. At the end of the experiment, the mice were sacrificed, and the colorectal was taken out to observe the changes in colorectal morphology in each group and observe the degree of lesions by HE staining. ELISA were used to detect the expressions of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), interleukin 4 (IL-4) and interleukin 10 (IL-10) in the supernatant of colorectal tissue homogenate.Immunohistochemistry and RT-qPCR were used to detect the expression of nuclear factor-κB P65 (NF-κB P65) protein and mRNA in colorectal tissues.Western blotting were used to detect the expression of toll-like receptor 4(TLR4) and myeloid differentiation factor 88(MyD88) in colorectal tissues. Results: Compared with the control group, the weight of the model mice was significantly reduced from the 6th day to the 14th day, the difference was statistically significant (P<0.05); compared with the model group, the FMT low and high dose group mice from the 10th day to the 14th day Compared with the control group, the expression of TNF-α, IL-6, NF-κB p65 mRNA and protein, TLR4, MyD88 in colorectal tissue of the model group were significantly higher Protein expression was significantly increased (P<0.05), IL-4 and IL-10 expression was significantly decreased (P<0.05), while IL-4 and IL-10 in high dose FMT group were significantly higher than those in model group, and other indexes were significantly decreased (P<0.05). Conclusion: FMT can alleviate the inflammatory response by inhibiting TLR4/MyD88/NF-κB signal pathway and play a therapeutic role in UC. |
|
View Full Text
View/Add Comment Download reader |
| Close |
|
|
|
