亚砷酸钠对皮肤细胞损伤的细胞生物学探讨

【作者】 吴顺华；

【导师】 刘开泰； 石玉瑚； 杨晓燕；

【作者基本信息】 新疆医科大学 ， 劳动卫生与环境卫生学， 2001， 硕士

【摘要】 目的：建立人表皮细胞（KC）、真皮成纤维细胞（HDFib）培养体系，探讨亚砷酸钠（NaAsO₂）对人两种皮肤细胞系一般生物学性状以及形态结构的影响。方法：通过直接细胞计数法和MTT还原法检测NaAsO₂对两种细胞系生长的影响；用光学显微镜、透射电镜研究NaAsO₂诱导人成纤维细胞转化和损伤的改变。结果：两种细胞系原代培养发现，成纤维细胞较表皮细胞易于培养。直接细胞计数法测得KC和HDFib的LC₅₀分别为8μM和20μM。用MTT法检测不同剂量的NaAsO₂与对照组相比较，其MTT吸光度值均有明显升高，具有统计学意义（p＜0.01）。低浓度（0.8μM、1.6μM）对表皮细胞有明显促增殖作用，较高浓度（＞3.2μM）时，细胞生长受到抑制。透射电镜观察到10μM NaAsO₂染毒的成纤维细胞各细胞器形态发生变化，线粒体肿胀，甚至出现空泡变性，高尔基体减少，滑面内质网扩张，粗面内质网减少。随时间延长，细胞体积增大胞浆膜损坏，出现核固缩，胞浆成网状，线粒体空泡变性，嵴消失，微丝不规则，微绒毛增多，甚至可见到网状的染色质结构。结论：NaAsO₂对体外培养的表皮细胞和成纤维细胞有增殖作用；10μM NaAsO₂可以诱导皮肤成纤维细胞损伤。更多还原

【Abstract】 IAbstractl Objective Arsenic is a potent human carcinogen to which there is wor1dv~ide exposure through natural contamination of food and drinking water sources. But the molecular mechanism of caicinogenesis is not clear With human dem~l cell culttu~ and molecular biologic methods, we explored the change of dermal cell h~juiy induced by sodium arsenite. Methods Count cell number, M?17 reduction assay, transmission elecironmicmscope were performed. Results The growth of human keratinocytes(KC) and human dem-ial fibroblast(HDF1b) all showed three phases, sta~ant phase is vely short(<24h).Wtth different concentration of sodium ar~nite, the two cells were markedly st黱ulated(p <0.01) .The ha]f lethal doses were respectively 8 L?M and 20 1-i M. The low level dose group (0.8 ii M.. 1.614 M) improved the ii~lifemtkx1 of keratinocytes through MTh reduction assay, but the high level dose group (above 3.214 M )can inhibit the cell growth. When the human deimal fibmblast was exposed 1014 M sodium arsenite,there were remarkable pathological changes of LdtmsWmcture, such as Mitochondzia swelling~ polyiibosonie dropping from rough endoplasmic reticulum endc鏻asmic reticulum expending in human dermal fibroblast cells et al. As culture time prolonging, the cells coming to die. Conclusion The proliferation of dennal cell may pby a critical role in the development of arseniasis caused by arsenic; 1014 M sodium arsenite can induce injuiy of human demial fibmblast in culture.更多还原