【作者】 孟冬冬；

【导师】 邓耀良；

【作者基本信息】 广西医科大学 ， 泌尿外科， 2011， 硕士

【摘要】 目的观察体外细胞培养体系中人肾小管上皮细胞与钙化性纳米微粒的相互作用,探讨钙化性纳米微粒对人肾小管上皮细胞的损伤机制,探讨钙化性纳米微粒在肾结石形成中的可能作用。方法体外建立人肾小管上皮细胞与钙化性纳米微粒共同培养体系,并使用NADPH氧化酶抑制剂apocynin进行干预处理。分别运用光镜、电镜观察人肾小管上皮细胞在与钙化性纳米微粒相互作用过程中的形态变化。实验分为四组,采用比色法测定24h后各组细胞培养液中乳酸脱氢酶、丙二醛、透明质酸的含量,同时检测细胞内NADPH氧化酶活性,采用RT-PCR测定各实验组细胞NADPH氧化酶亚基p22phox和p47phox mRNA的表达情况。结果钙化性纳米微粒在体外能导致人肾小管上皮细胞的形态学改变,通过电子显微镜观察到人肾小管上皮细胞能够吸附、吞噬钙化性纳米微粒。钙化性纳米微粒也能导致人肾小管上皮细胞NADPH氧化酶p22phox、p47phox亚基mRNA的表达明显增强,同时伴有细胞氧化应激(oxidative stress,OS)损伤,使用apocynin干预后上述基因表达减少,同时细胞的氧化应激损伤也减轻。结论在体外人肾小管上皮细胞具有粘附、吞噬钙化性纳米微粒的能力;钙化性纳米微粒能够引起人肾小管上皮细胞NADPH氧化酶的活性增强,导致细胞的氧化应激损伤。更多还原

【Abstract】 Objective To observe the interaction of the calcifying nanoparticles (CNP) with human renal tubular epithelial cells (HK-2) in vitro. To investigate the mechanisms of HK-2 damage incited by CNP, and to investigate the potential role of CNP in the formation of nephrolithiasis.Methods Human renal tubular epithelial cells were cultured with CNP in vitro, and the morphology changes was observed by light microscopy and electron microscopy. To investigate the oxidative stress, NADPH oxidase inhibitor apocynin was chosed as the intervention treatment. The levels of LDH, MDA, HA in the mediums and the activity of NADPH oxidase in human renal tubular epithelial cells were assessed after 24 hours. The expression of NADPH oxidase subunit p22phox and p47phox mRNA were detected by real-time quantitative PCR (RT-PCR).Results CNP can cause morphology changes of the HK-2, have observed the adhesion and phagocytosis of CNP by the HK-2 under TEM. After 24 hours, the level of LDH, MDA, HA were significantly different among the 4 groups (P <0.05).Conclusion HK-2 has abilities of adhering and phagocyting with CNP; and CNP can cause the damage induced by oxidative stress of HK-2.更多还原