脂多糖对大鼠骨髓间充质干细胞增殖的影响

Effects of lipopolysaccharide on proliferation of bone marrow stem cells in rats

目的观察不同浓度脂多糖(LPS)对体外培养的大鼠骨髓间充质干细胞(BMSCs)增殖的影响,并探讨其可能的机制。方法应用全骨髓贴壁法体外分离培养大鼠BMSCs,采用免疫细胞化学法鉴定其表面标志。将BMSCs分为对照组及不同浓度LPS干预组,采用噻唑蓝(MTT)比色法观察其增殖情况。结果采用全骨髓贴壁法获取的BMSCs形态呈长梭形,呈集落样生长,免疫细胞化学鉴定,CD44阳性,CD34、CD45阴性。MTT检测结果显示,与对照组(0.190±0.037)相比,0.01、0.1及1μg/mlLPS干预组吸光度(A)值均增高,其中0.01μg/mlLPS干预组(0.253±0.030)增高明显,差异有统计学意义(P<0.05)。而10、100μg/mlLPS干预组A值则较对照组下降,其中10μg/mlLPS干预组(0.159±0.042)明显降低,差异有统计学意义(P<0.05)。结论低浓度LPS可以提高BMSCs的增殖能力,但随着LPS浓度的增大,BMSCs增殖能力反而下降。

Objective To investigate the effects of lipopolysaccharide （LPS） with different concentrations on proliferation of bone marrow stem cells （BMSCs） in vitro, and its possible mechanism. Methods Rat BMSCs were isolated from the whole bone marrow and cultured invivo, lmmunocytoehemical technique was employed to identify the cells. MTT method was used to examine the proliferation of BMSCs which were divided into the control group and several LPS-treated groups. Results The BMSCs by whole bone man＇ow culture appeared as long spindle-shaped cells in colony-like growth. By immunocytochemistry, the BMSCs tested CD44positive, CD34- and CD45-negative. MTT7 showed that A value was higher in the groups treated with 0.01, 0.1 and 1.0 ug/ml LPS, and was lower in the groups treated with 10 and 100 ug/ml LPS, as compared with the controls （0.190±0.037）. The greatest increase and reduction in A values （0.253±0.030, 0.159±0.042） were seen in the 1.0 ug/ml and 10 ug/ml LPS-treated groups, respectively （both P〈0.05）. Conclusion The proliferation of BMSCs in vitro can be affected with different concentrations of lipopolysaeeharide. While low-level LPS may be stimulative, BMSC proliferation may be suppressed with higher levels of LPS.