NK细胞条件Cd226敲除小鼠的构建及其在溃疡性结肠炎中的作用的初步研究。

细胞与分子免疫学杂志 Pub Date : 2025-06-01
Jianchun Lyu, Zichan Guo, Yazhen Wang, Ziyan Chen, Zhengxiang Zhang, Lihua Chen
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引用次数: 0

摘要

目的利用Cre-loxP技术制备自然杀伤细胞(NK细胞)条件Cd226基因敲除小鼠并对其进行表征,探讨Cd226对NK细胞在减轻溃疡性结肠炎(UC)小鼠肠道炎症中的作用。方法通过Cre-loxP系统,将loxp侧链Cd226小鼠与Ncr1-Cre小鼠杂交,制备NK细胞条件Cd226基因敲除小鼠。采用聚合酶链反应(PCR)和琼脂糖凝胶电泳进行基因分型。葡聚糖硫酸钠(DSS)诱导建立UC模型。流式细胞术检测CD226在NK细胞上的表达水平及结肠组织中相关免疫细胞的浸润情况。苏木精-伊红(HE)染色评估结肠炎症程度。结果DNA凝胶电泳和流式细胞术证实成功产生NK细胞特异性Cd226敲除小鼠。条件敲除NK细胞中的Cd226后,UC小鼠模型的炎症得到缓解。流式细胞术结果显示外周血和结肠固有层NK细胞比例降低,HE染色显示炎症反应减弱。结论NK细胞特异性敲除Cd226可通过减少NK细胞数量和抑制其促炎功能减轻UC小鼠肠道炎症。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Construction of NK cell-conditional Cd226 knockout mice and preliminary investigation of their role in ulcerative colitis].

Objective To generate and characterize natural killer cell (NK cell)-conditional Cd226 gene knockout mice using Cre-loxP technology, and to explore the role of CD226 on NK cells in alleviating intestinal inflammation in a murine model of ulcerative colitis (UC). Methods NK cell-conditional Cd226 gene knockout mice were generated by crossing loxP-flanked Cd226 mice with Ncr1-Cre mice via the Cre-loxP system. Polymerase chain reaction (PCR) and agarose gel electrophoresis were used for genotyping. A UC model was established by dextran sulfate sodium (DSS) induction. Flow cytometry was performed to analyze CD226 expression levels on NK cells and the infiltration of related immune cells in colon tissues. Hematoxylin-eosin (HE) staining was performed to assess the degree of colonic inflammation. Results DNA gel electrophoresis and flow cytometry confirmed the successful generation of NK cell-specific Cd226 knockout mice. After conditional knockout of Cd226 in NK cells, inflammation in the UC mouse model was alleviated. Flow cytometry results showed a reduced proportion of NK cells in peripheral blood and the colon lamina propria, while HE staining demonstrated attenuated inflammatory responses. Conclusion Specific knockout of Cd226 in NK cells mitigates intestinal inflammation in UC mice by reducing NK cell numbers and inhibiting their pro-inflammatory functions.

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