本文選題：骨髓間充質(zhì)干細胞 + 巨噬細胞�。� 參考：《蘭州大學》2008年碩士論文

【摘要】： 目的探討在體外條件下骨髓間充質(zhì)干細胞(MSCs)對脂多糖(LPS)刺激后小鼠腹腔巨噬細胞活化及其功能的影響。 方法采用貼壁篩選法分離、純化BALB/c小鼠骨髓MSCs,用經(jīng)過高壓滅菌的4%巰基乙酸鈉溶液腹腔注射刺激,4天后收集小鼠腹腔巨噬細胞,依材料和方法所述分組,建立MSCs與巨噬細胞共培養(yǎng)體系。用LPS(終濃度1ug/ml)刺激巨噬細胞18小時后收集細胞培養(yǎng)上清,檢測培養(yǎng)體系中腫瘤壞死因子-α(TNF-α),轉(zhuǎn)化生長因子-β(TGF-β)和一氧化氮(NO)等炎癥性細胞因子分泌量的變化,同時在培養(yǎng)體系中加入大腸桿菌標準菌株(ATCC25922),共孵育24小時后用緩沖液洗滌,盡量除去未被吞噬的細菌后固定,采用瑞氏染色檢查巨噬細胞吞噬功能的變化。 結(jié)果用巰基乙酸鈉溶液腹腔注射刺激方法收集的小鼠腹腔巨噬細胞為炎癥性巨噬細胞,其培養(yǎng)上清中可檢測到少量細胞因子,經(jīng)LPS進一步刺激活化后培養(yǎng)上清中TNF-α和NO的含量明顯上升,分別為(147.41±37.13)pg/ml,(59.93±8.74)uM;而與MSCs共培養(yǎng)時,TNF-α顯著減少[(97.58±30.26)pg/ml,P=0.032],NO降到(50.90±29.48)uM,P＞0.05;當培養(yǎng)體系中存在MSCs上清時,即MPMs+LPS+MSC上清組中,TNF-α和NO進一步減少為(58.28±31.54)pg/ml,(-3.36±2.31)uM,P＜0.0005;然而,各組中TGF-β含量的差異無統(tǒng)計學意義,P＞0.05。MSCs對巨噬細胞活化后的吞噬率及吞噬指數(shù)沒有影響。 結(jié)論實驗首次發(fā)現(xiàn),MSCs可抑制LPS刺激后小鼠腹腔巨噬細胞的活化,而對其吞噬功能沒有影響。
[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- 偽 -TNF- 偽, 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- 偽 and no in supernatant stimulated by LPS increased significantly (147.41 鹵37.13 ng / ml, 59.93 鹵8.74 渭 M, respectively), while in co-cultured with MSCs, the content of TNF- 偽 decreased significantly [97.58 鹵30.26 ng / ml P0.032] no decreased to 50.90 鹵29.48 渭 m (P > 0.05). When the supernatant of MSCs existed in the culture system, the content of TNF- 偽 in the supernatant was significantly decreased to 50.90 鹵29.48 渭 M (P > 0.05), while in co-culture with MSCs, TNF- 偽 significantly decreased to 50.90 鹵29.48 渭 M (P > 0.05).That is, the levels of TNF- 偽 and no in MPMs LPS MSC supernatant group were further reduced to 58.28 鹵31.54 ng 路ml 路ml ~ (-1) -3.36 鹵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.
【學位授予單位】：蘭州大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R329.2

【引證文獻】

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