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1.610031 成都,西南交通大学生命科学与工程学院
2.400042 重庆,陆军特色医学中心野战外科研究部,创伤、烧伤与复合伤国家重点实验室,特殊环境战伤防治研究室
3.400042 重庆,陆军特色医学中心检验科
[李遂焰,E-mail:lisuiyan@swjtu.edu.cn LI Suiyan, E-mail: lisuiyan@swjtu.edu.cn
[严军,E-mail:13883092250@163.com YAN Jun, E-mail: 13883092250@163.com
收稿:2022-09-16,
修回:2022-10-14,
纸质出版:2023-03-30
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张雷廷, 夏雨, 余静, 等. Musculin影响小鼠骨髓来源树突状细胞生物学功能的初步研究[J]. 陆军军医大学学报, 2023,45(6):539-546.
Leiting ZHANG, Yu XIA, Jing YU, et al. Effect of Musculin on biological function of mouse bone marrow-derived dendritic cells[J]. Journal of Army Medical University, 2023, 45(6): 539-546.
张雷廷, 夏雨, 余静, 等. Musculin影响小鼠骨髓来源树突状细胞生物学功能的初步研究[J]. 陆军军医大学学报, 2023,45(6):539-546. DOI: 10.16016/j.2097-0927.202209153.
Leiting ZHANG, Yu XIA, Jing YU, et al. Effect of Musculin on biological function of mouse bone marrow-derived dendritic cells[J]. Journal of Army Medical University, 2023, 45(6): 539-546. DOI: 10.16016/j.2097-0927.202209153.
目的
2
探讨新型转录因子Musculin对小鼠骨髓来源树突状细胞(bone marrow-derived dendritic cells
BMDC)生物学功能的影响。
方法
2
获取野生型(wildtype
WT)和Musculin敲除(knockout
KO
-/-)小鼠骨髓细胞,用粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor
GM-CSF)和IL-4体外诱导BMDC,7 d后加入脂多糖(lipopolysaccharide
LPS),继续培养24 h;动态观察细胞形态变化,收集WT组和Musculin-/-组BMDC及上清,流式细胞术分析各组细胞表达CD11c、CD40、CD80、CD86、PD-L1和IAIE的比例,CCK-8法检测各组细胞刺激异源淋巴细胞增殖情况,并计算刺激指数(stimulation indexes
SI),ELISA法检测各组细胞上清中IL-1β、IL-6、IL-10、IL-12(P40)、IL-12(P70)和TNF-α的水平。
结果
2
与WT组比较,Musculin-/-组BMDC形态相似,纯度无明显差异;表面共刺激分子CD40、CD80和CD86的比例显著下降(
P
<
0.05),共抑制分子PD-L1和MHC-Ⅱ分子IAIE的比例也降低,但差异无统计学意义;刺激异源淋巴细胞后的数量和刺激指数上升,尤其是刺激5 d以后(
P
<
0.05);上清中促炎因子IL-1β、IL-6、IL-12(P40)、IL-12(P70)和TNF-α水平显著升高(
P
<
0.05),但抗炎因子IL-10的水平变化不明显。
结论
2
Musculin可能增强BMDC介导的免疫功能并抑制炎症反应,提示Musculin能够调控BMDC的生物学功能。
Objective
2
To investigate the effect of a novel transcription factor
Musculin
on the biological functions of bone marrow-derived dendritic cells (BMDC) in mice.
Methods
2
Bone marrow cells from wildtype (WT) and Musculin knockout (KO
-/-) mice were obtained and cultured with granulocyte-macrophage colony stimulating factor (GM-CSF) and IL-4 so as to induce
BMDC
in vitro
. After 7 d
the BMDC were added with lipopolysaccharide (LPS) and continued to culture for 24 h. Besides the cellular morphological changes were observed dynamically
BMDC and the supernatants were collected from the WT and Musculin-/-cells. Flow cytometry was used to analyze the proportion of positive cells of CD11c
CD40
CD80
CD86
PD-L1 and IAIE in each group. CCK-8 assay was adopted to detect the cellular proliferation for stimulating heterologous lymphocytes and the different stimulation indexes (SI) were calculated respectively. ELISA was applied to detect the supernatant contents of IL-1β
IL-6
IL-10
IL-12 (P40)
IL-12 (P70) and TNF-α in each group.
Results
2
The BMDC from the Musculin-/-group had similar morphology and no significant difference in purity
decreased percentages of co-stimulatory molecules such as CD40
CD80 and CD86 (
P
<
0.05)
and reduced percentages of co-inhibitory molecules PD-L1 and major histocompatibility complex class Ⅱ (MHC-Ⅱ) IAIE though no significant differences when compared with those of WT group. Moreover
the number of heterologous lymphocytes and related SI after of mixed lymphocyte reaction (MLR) were obvious higher in the Musculin-/-group than the WT group
especially after 5 days' stimulation (
P
<
0.05). In addition
the supernatant levels of proinflammatory factors including IL-1β
IL-6
IL-12 (P40)
IL-12 (P70) and TNF-α were significantly increased in the Musculin-/-group than the WT group (
P
<
0.05)
but no such changes were observed in anti-inflammatory factor IL-10 level between the 2 groups.
Conclusion
2
Musculin may enhance the immune function mediated by BMDC and suppress inflammatory reaction
suggesting that it can regulate the biological functions on BMDC.
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