[1]孙中美,李军祥,路琼琼,等.清肠温中方联合粪菌移植对溃疡性结肠炎小鼠肠黏膜免疫的调控作用[J].西部中医药,2024,37(02):9-13.[doi:10.12174/j.issn.2096-9600.2024.02.03]
　SUN Zhongmei,LI Junxiang,LU Qiongqiong,et al.Regulatory Effects of Qingchang Wenzhong Prescription and Fecal Microbiota Transplantation on Intestinal Mucosal Immunity in Mice with Ulcerative Colitis[J].Western Journal of Traditional Chinese Medicine,2024,37(02):9-13.[doi:10.12174/j.issn.2096-9600.2024.02.03]
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清肠温中方联合粪菌移植对溃疡性结肠炎小鼠肠黏膜免疫的调控作用
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《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
37
期数:
2024年02期
页码:
9-13
栏目:
基础研究
出版日期:
2024-02-15

文章信息/Info

Title:
Regulatory Effects of Qingchang Wenzhong Prescription and Fecal Microbiota Transplantation on Intestinal Mucosal Immunity in Mice with Ulcerative Colitis
作者:
孙中美1,2, 李军祥1, 路琼琼1, 丁庞华1, 姜慧1, 施晓军1, 毛堂友1
1.北京中医药大学东方医院，北京 100029
2.天津市南开医院，天津 300102
Author(s):
SUN Zhongmei1,2, LI Junxiang1, LU Qiongqiong1, DING Panghua1, JIANG Hui1, SHI Xiaojun1, MAO Tangyou1
1.Dongfang Hospital of Beijing University of Chinese Medicine, Beijing 100029, China
2.Tianjin Hospital of ITCWM Nankai Hospital, Tianjin 300102, China
关键词:
结肠炎; 溃疡性; 清肠温中方; 粪菌移植; 肠黏膜免疫; T细胞; 小鼠; 动物实验
Keywords:
colitis;  ulcerative; prescription; fecal microbiota transplantation; intestinal mucosal immunity; T cells; mice; zoopery
分类号:
R516.1
DOI:
10.12174/j.issn.2096-9600.2024.02.03
文献标志码:
A
摘要:
目的探究清肠温中方联合粪菌移植对溃疡性结肠炎小鼠肠黏膜免疫应答的调控作用。 方法将30只健康的SPF级雌性C57BL/6小鼠按照1∶4的比例随机分为空白组（6只）、干预组（24只）。空白组小鼠全程自由饮用无菌水，并自第8天起给予无菌水灌胃，同时收集新鲜粪便，便制作粪便滤液；干预组自由饮用2.5%葡聚糖硫酸钠（dextran sulfate sodium，DSS）溶液7天建立UC模型后，随机分为模型组、清肠温中方组、粪菌移植组、清肠温中方联合粪菌移植组（联合组），各组干预7天。观察小鼠一般情况，测量体质量、检测粪便潜血、记录粪便性状，计算疾病活动指数。干预结束后处死小鼠，结肠石蜡切片HE染色，光镜下观察组织病理学改变；取脾脏，运用流式细胞术检测γδT细胞、CD4+T细胞；取小鼠结肠组织，运用RT-qPCR检测TLR2 mRNA、pIgR mRNA的表达，Elisa检测sIgA的含量。 结果与空白组小鼠相比，DSS诱导结肠炎小鼠呈现明显的肠道炎症，主要表现为不同程度的便血、体质量下降、大便稀溏等；而当DSS停用后，各项临床表征开始缓解，各干预组小鼠呈现相对快速地恢复过程，疾病活动指数及组织病理学评分较模型组小鼠明显降低（P＜0.05）。对肠黏膜免疫相关指标检测结果发现，模型组小鼠γδT细胞、CD4+T细胞含量及结肠TLR2 mRNA表达明显升高（P＜0.05或P＜0.01），结肠pIgR mRNA、结肠sIgA表达较空白组明显降低（P＜0.05或P＜0.01）。经过1周的治疗后，各干预组小鼠γδT细胞、CD4+T细胞、结肠TLR2 mRNA表达明显降低（P＜0.05），结肠pIgR mRNA、结肠sIgA表达明显升高（P＜0.05或P＜0.01）；其中联合组改善肠黏膜损伤方面更为明显。 结论清肠温中方联合粪菌移植可通过影响肠黏膜免疫应答，改善溃疡性结肠炎小鼠的肠道炎症，促进肠黏膜损伤的修复。
Abstract:
ObjectiveTo survey the regulatory effects of Qingchang Wenzhong prescription combined with fecal microbiota transplantation on intestinal mucosal immunity in mice suffering ulcerative colitis (UC). MethodsThirty healthy SPF-grade female C57BL/6 mice were randomized into the blank group (six ones) and the intervention group (24 ones) according to the ratio of 1:4. The mice in the blank group were allowed to drink sterile water throughout the day and began to accept intragastric administration of sterile water since the eighth day, meanwhile, fresh faeces were collected to prepare faecal filtrate; UC models were established after the intervention group had freely drank 2.5% DSS solutions for seven days, and they were randomized into the model group, Qingchang Wenzhong prescription group, fecal microbiota transplantation group, the combination group (Qingchang Wenzhong prescription and fecal microbiota transplantation), intervened for seven days. To observe general conditions of the mice, to detect body mass and faecal occult blood, to record fecal character and to calculate disease activity index (DAI). The mice were sacrificed by the end of intervention, to observe histopa-thological changes by light microscope after paraffin sections of the colon were processed by HE staining; the spleen was taken out to detect the expressions of γδT cells and CD4+T cells using flow cytometry; RT-qPCR method was used to measure the expressions of TLR2 mRNA and plgR mRNA after getting out the colon, ELISA was applied to test the contents of sIgA. ResultsCompared with the mice in the blank group, DSS-induced UC mice showed marked intestinal inflammation, mainly manifesting different degrees of hematochezia, body weight loss and loose stool; when DSS was withdrew, clinical symptoms began to relieve, the mice in different intervention groups presented relatively rapid recovery, DAI and histopathological scores were reduced noticeably than these of the model group (P<0.05). The detection results of intestinal mucosal immune-related indicators found that the contents of γδT cells and CD4+T cells and the expressions of TLR2 mRNA in the colon were raised remarkably in the model group (P<0.05 or P<0.01), the expressions of pIgR mRNA and sIgA in the colon were lowered than these of the blank group (P<0.05 or P<0.01). After one week of the treatment, the expressions of γδT cells, CD4+T cells and colonic TLR2 mRNA were clearly decreased in the intervention groups (P<0.05), the expressions of pIgR mRNA and sIgA were obviously lifted in the colon (P<0.05 or P<0.01); among them, the improvements of intestinal mucosal injury were the most apparent in the combination group. ConclusionQingchang Wenzhong prescription combined with fecal microbiota transplantation could improve intestinal inflammation in UC mice and promote the repair of intestinal mucosal injury by affecting intestinal mucosal immune response.

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备注/Memo

备注/Memo:
孙中美（1992—），女，博士学位，医师。研究方向：溃疡性结肠炎的中医药防治。国家自然科学基金青年基金（82374411）；北京市自然科学基金青年项目（7194294）；中华中医药学会（2022-2024年度）青年人才托举工程项目（CACM-2022-QNRC2-A02）。
更新日期/Last Update: 2024-02-15