巨噬细胞在脂多糖诱导肠上皮细胞凋亡中的作用

doi:10.12092/j.issn.1009-2501.2019.10.009

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (10): 1142-1146.doi: 10.12092/j.issn.1009-2501.2019.10.009

巨噬细胞在脂多糖诱导肠上皮细胞凋亡中的作用

曹迎亚，陈群，王箴，吴敬医，姜小敢，金孝岠，鲁卫华

皖南医学院弋矶山医院麻醉与重症医学科，芜湖 241001，安徽

收稿日期:2018-08-19 修回日期:2018-09-13 出版日期:2019-10-26 发布日期:2019-10-28
通讯作者: 鲁卫华，男，硕士，主任医师，主要从事重要脏器的功能维护及围术期并发症防治研究。 Tel：0553-5739735 E-mail：lwh683@126.com
作者简介:曹迎亚，女，硕士，医师，研究方向:脓毒症肠屏障功能。 Tel：15055324662 E-mail：caoyingya1990@126.com
基金资助:
安徽省自然科学基金青年项目（1908085QH360）；安徽省高校拔尖人才项目（gxbjZD19）；安徽省学术和技术带头人后备人选科研项目（2017H145）；皖南医学院中青年科研基金（WK2016020）

Role of macrophages in LPS-induced apoptosis of intestinal epithelial cells

CAO Yingya, CHEN Qun, WANG Zhen, WU Jingyi, JIANG Xiaogan, JIN Xiaoju, LU Weihua

Department of Anesthesiology and Critical Care Medicine, Yijishan Hospital, Wannan Medical College, Wuhu 241001, Anhui, China

Received:2018-08-19 Revised:2018-09-13 Online:2019-10-26 Published:2019-10-28

摘要/Abstract

摘要：

目的:评价巨噬细胞在脂多糖（LPS）诱导肠上皮细胞凋亡中的作用及机制。方法:根据培养基不同分为普通培养基组（R组）和巨噬细胞条件培养基组（M组），将对数生长期的肠上皮细胞NCM460接种到六孔板中，再向其中加入不同浓度（0，0.1，1，10，30 μg/mL）的LPS处理24 h，收集细胞染色后用流式细胞分析仪检测细胞凋亡情况。同时收集细胞培养液上清，用ELISA法检测炎症因子TNF-α、IL-6水平。Western blot检测IκB-α蛋白水平。结果:相同培养基中，随着LPS处理浓度增加，细胞培养液上清中炎症因子TNF-α、IL-6水平及细胞凋亡率逐渐增加（P<0.05）。同等剂量的LPS刺激NCM460细胞，M组细胞培养液上清中炎症因子TNF-α、IL-6水平升高程度及细胞凋亡率增加较R组明显（P<0.05），M组IκB-α蛋白水平也较R组下调。结论:巨噬细胞在LPS诱导的肠上皮细胞凋亡中发挥促进作用，其机制可能与NF-κB活化释放大量炎症因子有关。

关键词: 脂多糖类, 巨噬细胞, 细胞凋亡, 肠上皮细胞

Abstract:

AIM: To evaluate the role and mechanism of macrophages on lipopolysaccharide (LPS)-induced apoptosis of intestinal epithelial cells. METHODS: NCM460 intestinal epithelial cells were incubated in regular culture medium (group R) or macrophage conditioned medium (group M) in 6-well plates. Different concentrations (0, 0.1, 10, 30 μg/mL) of LPS was added to the medium of NCM460 cells for 24 hours, then the cells were collected for Annexin V/7AAD staining, following with flow cytometry analysis. The cell culture supernatant was also collected to detect the TNF-α and IL-6 levels by enzyme-linked immunosorbent assay. Western blot was used to measure the activity of NF-κB signaling pathway. RESULTS:In the same culture medium, with the increase of LPS concentration, the levels of inflammatory factors TNF-α, IL-6 and apoptotic rate in the supernatant of cell culture medium increased gradually (P<0.05). Compared with group R, the percentage of apoptotic cells were significantly increased in group M when cells were treated with equal concentrations of LPS. Compared with NCM460 cells stimulated by LPS at the same dose, the apoptotic rate and the levels of inflammatory factors TNF-α and IL-6 in supernatant of M group were higher than those of R group (P<0.05). Compared with group R, the expression levels of IκB-α protein was down-regulated.CONCLUSION: Macrophages play an important role in LPS-induced apoptosis of intestinal epithelial cells. The mechanism may be related to the release of inflammatory cytokines by activation of NF-κB signaling pathway.

Key words: lipopolysaccharides, macrophages, apoptosis, enterocyte

中图分类号:

R965.2

曹迎亚，陈群，王箴，吴敬医，姜小敢，金孝岠，鲁卫华. 巨噬细胞在脂多糖诱导肠上皮细胞凋亡中的作用[J]. 中国临床药理学与治疗学, 2019, 24(10): 1142-1146.

CAO Yingya, CHEN Qun, WANG Zhen, WU Jingyi, JIANG Xiaogan, JIN Xiaoju, LU Weihua. Role of macrophages in LPS-induced apoptosis of intestinal epithelial cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(10): 1142-1146.

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巨噬细胞在脂多糖诱导肠上皮细胞凋亡中的作用

Role of macrophages in LPS-induced apoptosis of intestinal epithelial cells

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