中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (31): 4962-4966.doi: 10.12307/2022.738

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

脂多糖干预小鼠骨髓来源巨噬细胞的凋亡

郑美洁1,杨玮哲1,刘佳林1,韩祥祯1,2,周琦琪1,2,何惠宇1,2   

  1. 1新疆医科大学第一附属医院(附属口腔医院)口腔修复科,新疆维吾尔自治区乌鲁木齐市   830054;2新疆维吾尔自治区口腔医学研究所,新疆维吾尔自治区乌鲁木齐市   830054
  • 收稿日期:2021-10-08 接受日期:2021-11-25 出版日期:2022-11-08 发布日期:2022-04-25
  • 通讯作者: 何惠宇,主任医师,教授,博士,博士生导师,硕士生导师,新疆医科大学第一附属医院(附属口腔医院)口腔修复科,新疆维吾尔自治区乌鲁木齐市 830054;新疆维吾尔自治区口腔医学研究所,新疆维吾尔自治区乌鲁木齐市 830054
  • 作者简介:郑美洁,女,1996年生,山东省德州市人,汉族,在读硕士,主要从事口腔修复学及组织工程基础研究。
  • 基金资助:
    新疆维吾尔自治区科技支疆项目(2020E02133),项目负责人:韩祥祯;新疆维吾尔自治区研究生科研创新项目(XJ2021G196),项目负责人:刘佳林

Apoptosis of mouse bone marrow-derived macrophages induced by lipopolysaccharide

Zheng Meijie1, Yang Weizhe1, Liu Jialin1, Han Xiangzhen1, 2, Zhou Qiqi1, 2, He Huiyu1, 2   

  1. 1Department of Prosthodontics, First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Received:2021-10-08 Accepted:2021-11-25 Online:2022-11-08 Published:2022-04-25
  • Contact: He Huiyu, Chief physician, Professor, MD, Doctoral supervisor, Master’s supervisor, Department of Prosthodontics, First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Zheng Meijie, Master candidate, Department of Prosthodontics, First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    Science and Technology Support Project of Xinjiang Uygur Autonomous Region, No. 2020E02133 (to HXZ); Postgraduate Research and Innovation Project of Xinjiang Uygur Autonomous Region, No. XJ2021G196 (to LJL)

摘要:

文题释义:
细胞凋亡:是调节细胞存活性以维持机体稳态的一种调节方式,有区别于细胞死亡与焦亡,其分为内源性途径(即线粒体途径)和外源性途径(即死亡受体途径),其特征性符号也随其凋亡而凸显,是目前肿瘤学及免疫学等研究的热点。
异物反应:当植入物等修复材料植入机体后,免疫系统应对外来异物所做出的应答反应,主要由材料表面的巨噬细胞及异物巨细胞介导,其长久的存在可能会导致部分植入物过早的裂解,使得修复治疗面临失败,因而异物反应的调节与组织再生速率匹配问题成为研究的关键。

背景:巨噬细胞是机体应对外界刺激的一道免疫防线,任何生物材料植入体内后的相关反应均与巨噬细胞介导的免疫反应有联系。
目的:探讨脂多糖对小鼠骨髓来源巨噬细胞凋亡的影响。
方法:体外分离培养C57BL/6小鼠骨髓来源巨噬细胞,分别加入含0(对照),10,100,1 000 µg/L脂多糖的DMEM完全培养基。培养24 h后,利用流式细胞仪检测细胞凋亡率,Hoechst染色观察细胞形态,RT-PCR检测白细胞介素1、白细胞介素6、肿瘤坏死因子α及凋亡相关因子Bax、P53的mRNA表达。
结果与结论:①各质量浓度脂多糖组的细胞凋亡率均高于对照组(P < 0.05),其中100,1 000 µg/L脂多糖组细胞凋亡率高于10 µg/L脂多糖组(P < 0.05);②Hoechst染色显示,对照组细胞核正常,细胞发生凋亡时染色质浓缩;各质量浓度脂多糖组细胞发生不同程度凋亡,凋亡细胞的细胞核呈致密浓染或碎块状致密浓染,颜色稍有些发白,尤以1 000 µg/L最明显;③RT-PCR检测显示,各质量浓度脂多糖组细胞的白细胞介素1、白细胞介素6、肿瘤坏死因子α、Bax及P53 mRNA表达均高于对照组(P < 0.01或P < 0.001),且随着脂多糖质量浓度的升高,白细胞介素1、白细胞介素6、肿瘤坏死因子α mRNA表达呈先升高后降低的趋势,Bax、P53 mRNA表达呈升高趋势;④结果表明,脂多糖可诱导骨髓来源巨噬细胞的凋亡。

https://orcid.org/0000-0001-6605-1490 (郑美洁)

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关键词: 植入体, 骨髓, 巨噬细胞, 脂多糖, 凋亡, 骨组织工程

Abstract: BACKGROUND: Macrophage is an immune defense line in response to external stimuli. The related reaction of any biological material implanted in vivo is related to the immune response mediated by macrophage.  
OBJECTIVE: To investigate the effect of lipopolysaccharide on the apoptosis of bone marrow-derived macrophages in mice.
METHODS:  The bone marrow-derived macrophages of C57BL/6 mice were isolated in vitro and cultured with DMEM containing 0 (control), 10, 100, and 1 000 µg/L lipopolysaccharide for 24 hours. Apoptotic rate was identified by flow cytometry. Hoechst staining was used to observe the cell morphology. RT-PCR was used to detect the mRNA expression levels of interleukin 1, interleukin 6, tumor necrosis factor alpha, and apoptosis-related factors Bax and P53.  
RESULTS AND CONCLUSION: (1) The apoptotic rate of the lipopolysaccharide group at each mass concentration was higher than that of the control group (P < 0.05), and the apoptotic rates of the 100, 1 000 µg/L lipopolysaccharide groups were higher than that of the 10 µg/L lipopolysaccharide group (P < 0.05). (2) Hoechst staining showed that the nucleus of the control group was normal, and the chromatin was concentrated when the cells underwent apoptosis. The cells in the lipopolysaccharide group of various mass concentrations experienced different degrees of apoptosis, and the nuclei of the apoptotic cells were densely stained or fragmented densely stained; the color was slightly whitish, especially at 1 000 µg/L. (3) RT-PCR detection showed that the expression levels of interleukin 1, interleukin 6, tumor necrosis factor α, Bax, and P53 mRNA in the cells of the lipopolysaccharide group of various mass concentrations were higher than those of the control group (P < 0.01 or P < 0.001). With the increased concentration of lipopolysaccharide, the expression levels of interleukin 1, interleukin 6, and tumor necrosis factor α mRNA showed a trend of first increasing and then decreasing, and the expression of Bax and P53 mRNA showed an increasing trend. (4) Results have confirmed that lipopolysaccharide can induce the apoptosis of bone marrow-derived macrophages.

Key words: implant, bone marrow, macrophage, lipopolysaccharide, apoptosis, bone tissue engineering

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