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骨髓间充质干细胞对小鼠腹腔巨噬细胞活化及功能影响的实验研究

[Experimental study on influence of bone marrow mesenchymal stem cells on activation and function of mouse peritoneal macrophages].

作者信息

Yang Yi-Wu, Bai Hai, Wang Cun-Bang, Lin Mei, Wu Ling-Qi

机构信息

Department of Hematology, Lanzhou General Hospital, Lanzhou Military Area Command, Lanzhou 730050, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2008 Aug;29(8):540-3.

Abstract

OBJECTIVE

To explore the influence of bone marrow (BM) mesenchymal stem cells (MSCs) on macrophage activation after lipopolysaccharide (LPS) stimulation.

METHODS

Mouse BM MSCs were isolated and purified by adherence screening, and mouse peritoneal macrophages (MPM) were collected by sodium thioglycollate peritoneal injection, and the co-culture system was established by planting macrophages on the MSCs monolayer. The grouping of experiments: group A: MPM; group B: MPM + LPS; group C: MPM + LPS + MSC; group D: MPM + LPS + MSC supernatant. Cell culture supernatants were collected to detect the changes of TNF-alpha/TGF-beta and nitrogen monoxide (NO) after stimulating macrophages with LPS for 18 hours. At the same time Escherichia coli standard strain (ATCC25922) was added into the culture system and incubated for another 24 hours, macrophages were stained and phagocytosis were examined.

RESULTS

The concentrations of TNF-alpha and NO in culture supernatants were increased significantly to (147.4 +/- 37.1) pg/ml and (59.9 +/- 8.7) micromol/L respectively after macrophage activation, however, at the present of MSC, the concentration of TNF-alpha was dramatically decreased [(97.6 +/- 30.3) pg/ml, P = 0.032], and the concentration of NO was decreased to (50.9 +/- 29.5) micromol/L (P > 0.05). The concentrations of TNF-alpha and NO were further decreased after addition of MSC supernatants [(58.3 +/- 31.5) pg/ml and (-3.4 +/- 2.3) micromol/L respectively, P < 0.01]. There was no change in the phagocytic rate and phagoindex of macrophages after activation.

CONCLUSIONS

MSCs can inhibit the activation of mouse peritoneal macrophages after stimulating with LPS but has no influence on the phagocytosis.

摘要

目的

探讨骨髓间充质干细胞(MSCs)对脂多糖(LPS)刺激后巨噬细胞活化的影响。

方法

通过贴壁筛选分离纯化小鼠骨髓间充质干细胞,经巯基乙酸钠腹腔注射收集小鼠腹腔巨噬细胞(MPM),将巨噬细胞种植于间充质干细胞单层上建立共培养体系。实验分组:A组:MPM;B组:MPM + LPS;C组:MPM + LPS + MSC;D组:MPM + LPS + MSC上清液。收集细胞培养上清液,检测LPS刺激巨噬细胞18小时后肿瘤坏死因子-α/转化生长因子-β及一氧化氮(NO)的变化。同时向培养体系中加入大肠埃希菌标准菌株(ATCC25922),继续培养24小时,对巨噬细胞进行染色并检测吞噬功能。

结果

巨噬细胞活化后,培养上清液中肿瘤坏死因子-α和NO浓度显著升高,分别达到(147.4±37.1)pg/ml和(59.9±8.7)μmol/L,然而,在有间充质干细胞存在时,肿瘤坏死因子-α浓度显著降低[(97.6±30.3)pg/ml,P = 0.032],NO浓度降至(50.9±29.5)μmol/L(P>0.05)。加入间充质干细胞上清液后,肿瘤坏死因子-α和NO浓度进一步降低[分别为(58.3±31.5)pg/ml和(-3.4±2.3)μmol/L,P<0.01]。巨噬细胞活化后吞噬率和吞噬指数无变化。

结论

间充质干细胞可抑制LPS刺激后小鼠腹腔巨噬细胞的活化,但对吞噬功能无影响。

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