间充质干细胞对巨噬细胞活化及其功能影响的实验研究

发布时间：2018-04-17 05:24

本文选题：骨髓间充质干细胞 + 巨噬细胞　；参考：《兰州大学》2008年硕士论文

【摘要】： 目的探讨在体外条件下骨髓间充质干细胞(MSCs)对脂多糖(LPS)刺激后小鼠腹腔巨噬细胞活化及其功能的影响。方法采用贴壁筛选法分离、纯化BALB/c小鼠骨髓MSCs,用经过高压灭菌的4%巯基乙酸钠溶液腹腔注射刺激,4天后收集小鼠腹腔巨噬细胞,依材料和方法所述分组,建立MSCs与巨噬细胞共培养体系。用LPS(终浓度1ug/ml)刺激巨噬细胞18小时后收集细胞培养上清,检测培养体系中肿瘤坏死因子-α(TNF-α),转化生长因子-β(TGF-β)和一氧化氮(NO)等炎症性细胞因子分泌量的变化,同时在培养体系中加入大肠杆菌标准菌株(ATCC25922),共孵育24小时后用缓冲液洗涤,尽量除去未被吞噬的细菌后固定,采用瑞氏染色检查巨噬细胞吞噬功能的变化。结果用巯基乙酸钠溶液腹腔注射刺激方法收集的小鼠腹腔巨噬细胞为炎症性巨噬细胞,其培养上清中可检测到少量细胞因子,经LPS进一步刺激活化后培养上清中TNF-α和NO的含量明显上升,分别为(147.41±37.13)pg/ml,(59.93±8.74)uM;而与MSCs共培养时,TNF-α显著减少[(97.58±30.26)pg/ml,P=0.032],NO降到(50.90±29.48)uM,P＞0.05;当培养体系中存在MSCs上清时,即MPMs+LPS+MSC上清组中,TNF-α和NO进一步减少为(58.28±31.54)pg/ml,(-3.36±2.31)uM,P＜0.0005;然而,各组中TGF-β含量的差异无统计学意义,P＞0.05。MSCs对巨噬细胞活化后的吞噬率及吞噬指数没有影响。结论实验首次发现,MSCs可抑制LPS刺激后小鼠腹腔巨噬细胞的活化,而对其吞噬功能没有影响。
[Abstract]:Objective to investigate the effects of bone marrow mesenchymal stem cells (MSCs) on the activation and function of murine peritoneal macrophages stimulated by lipopolysaccharide (LPS) in vitro.Methods MSCs of BALB/c mice bone marrow were isolated and purified by adherent screening method. Peritoneal macrophages were collected by intraperitoneal injection of 4% sodium mercaptoacetate solution after high pressure sterilization for 4 days, and the macrophages were grouped according to the materials and methods mentioned above.The co-culture system of MSCs and macrophage was established.The supernatants of macrophages were collected after stimulated with LPS-1ug-ml for 18 hours, and the secretion of inflammatory cytokines such as tumor necrosis factor- 伪 -TNF- 伪, transforming growth factor- 尾 (TGF- 尾) and nitric oxide (no) in the culture system were detected.At the same time, the standard Escherichia coli strain ATCC 25922 was added to the culture system. After incubation for 24 hours, the bacteria were washed with buffer solution, and the bacteria that had not been phagocytized were removed and fixed. The phagocytic function of macrophages was detected by Reich staining.Results the peritoneal macrophages collected by intraperitoneal injection of sodium mercaptoacetate were inflammatory macrophages, and a small amount of cytokines could be detected in the supernatants.The contents of TNF- 伪 and no in supernatant stimulated by LPS increased significantly (147.41 卤37.13 ng / ml, 59.93 卤8.74 渭 M, respectively), while in co-cultured with MSCs, the content of TNF- 伪 decreased significantly [97.58 卤30.26 ng / ml P0.032] no decreased to 50.90 卤29.48 渭 m (P > 0.05). When the supernatant of MSCs existed in the culture system, the content of TNF- 伪 in the supernatant was significantly decreased to 50.90 卤29.48 渭 M (P > 0.05), while in co-culture with MSCs, TNF- 伪 significantly decreased to 50.90 卤29.48 渭 M (P > 0.05).That is, the levels of TNF- 伪 and no in MPMs LPS MSC supernatant group were further reduced to 58.28 卤31.54 ng 路ml 路ml ~ (-1) -3.36 卤2.31 渭 M ~ (-1) (P < 0.0005), but there was no significant difference in the content of TGF- 尾 in each group (P > 0.05.MSCs) on the phagocytosis rate and phagocytic index after activation of macrophages.Conclusion LPS can inhibit the activation of peritoneal macrophages in mice for the first time, but has no effect on phagocytosis.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R329.2

【引证文献】