利用显微技术对体外诱导巨噬细胞与肾结石晶体相互作用的形态学研究

发布时间：2018-03-08 03:35

本文选题：巨噬细胞　切入点：肾结石晶体　出处：《广西医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的:研究体外细胞培养体系中人巨噬细胞U937与肾结石晶体的相互作用，观察细胞与晶体反应的形态学改变，并探讨巨噬细胞在肾结石形成中的可能作用。方法:体外利用PMA（Phorbol-12-myristate-13-acetate，，PMA)对人单核细胞株U937进行诱导，激活分化为巨噬细胞后，并使用不同种类的肾结石晶体与之进行共培养。分别运用倒置相差显微镜、扫描电子显微镜及活细胞成像技术—显微延时摄影静态及动态地观察对细胞与晶体的相互作用。根据细胞与不同种类晶体共培养的时段不同分组，采用茜素红S—氯化十六烷基吡啶钙盐半定量测定法测定晶体-细胞反应后细胞基质的钙盐沉积程度。结果:光镜、扫描电镜下可见巨噬细胞吸附、吞噬晶体。活细胞成像技术—显微延时摄影捕捉到细胞吸附、吞噬、溶解及转运晶体的动态过程。茜素红S—氯化十六烷基吡啶钙盐半定量测定法：羟基磷灰石（hydroxyapatite，HAP）与一水草酸钙（calcium oxalate monohydrate，COM）混合实验组与其它组在相同时段细胞基质的钙盐沉积程度差别有统计学意义（P＜0.05），24小时组的程度最高；HAP组与空白对照组在相同时段差别有统计学意义（P＜0.05），与COM组差别无统计学意义（P＞0.05）。空白对照组与COM组在相同时段差别无统计学意义（P＞0.05）；各晶体实验组12小时组与24小时组后差别有统计学意义（P＜0.05）,6小时组分别与另外两个时段组差别无统计学意义（P＞0.05）；不同实验组不同时段不存在交互作用（P＞0.05）。结论：巨噬细胞具有粘附、吞噬及转运肾结石晶体的能力。HAP可能存在介导巨噬细胞增强吞噬HAP和COM的作用；巨噬细胞可能通过吞噬HAP后启动吞噬COM。
[Abstract]:Aim: to study the interaction between human macrophage U937 and renal stone crystals in vitro, observe the morphological changes of cell and crystal reaction, and explore the possible role of macrophages in the formation of renal calculi. Methods: human mononuclear cell line U937 was induced by PMA-Phorbol-12-Eritrestate-13-acetatePMA in vitro, then activated and differentiated into macrophages and co-cultured with different kinds of kidney stone crystals. Scanning electron microscopy and living cell imaging techniques-static and dynamic observation of the interaction between cells and crystals by time-delay photography. The calcium deposition degree of cell matrix after crystal-cell reaction was determined by alizarin red S-cetylpyridine calcium salt semi-quantitative method. Results: under light microscope, macrophage adsorption and phagocytosis were observed under scanning electron microscope. Dynamic process of dissolution and transport of crystals. Semi-quantitative determination of alizarin red S- cetylpyridine chloride calcium salt: hydroxyapatite HAP) and calcium oxalate monohydrate monohydrate monohydrate. There was significant difference in sedimentation degree between HAP group and blank control group at the same time period (P < 0.05), but there was no significant difference between HAP group and COM group (P > 0.05). The difference between blank control group and COM group was at the same time. There was no statistical significance (P > 0.05), there was no significant difference between the experimental group and the other two groups after 12-hour group and 24-hour group (P < 0.05), there was no significant difference between the other two groups (P > 0.05), and there was no interaction between the experimental group and the other two groups (P > 0.05). Conclusion: macrophages have adhesion. The ability of phagocytosis and transport of renal stone crystals. HAP may mediate macrophages to enhance phagocytosis of HAP and COM, and macrophages may initiate phagocytosis of HAP and COM by phagocytosis of HAP.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692.4

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