摘要
目的探讨金属硫蛋白(MT)-1、2在慢性水砷暴露小鼠肝损伤中的作用。方法将健康清洁级雄性昆明种小鼠随机分成对照组和暴露组,对照组给予普通饲料和自来水(可自由饮用);暴露组给予普通饲料和300mg/L的亚砷酸钠溶液(可自由饮用)。在暴露前,采用150mg/L亚砷酸钠溶液以自由饮用的方式喂养暴露组小鼠,17周后小鼠未出现死亡。染毒10个月后,检测小鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)的活力和球蛋白(Glb)的含量。采用TRIzol-酚-氯仿一步法提取总RNA,采用紫外分光光度法测定RNA在波长为260和280nm时的吸光度值(A260、A280),以A260/A280鉴定RNA的纯度。以实时荧光定量PCR(RT-PCT)技术测定小鼠肝组织中MT-1和MT-2mRNA。并进行肝组织病理学观察。结果对照组小鼠状态良好,毛发光泽;暴露组小鼠毛发粗糙,精神萎靡,动作迟钝,易激惹,进食、进水量明显减少。对照组小鼠体重为(39.10±3.48)g,高于暴露组[(14.00±1.49)g],差异有统计学意义(P<0.05)。与对照组比较,暴露组小鼠血清中ALT、AST的活力和Glb含量较高,肝组织中MT-1和MT-2mRNA的含量较低,差异均有统计学意义(P<0.05)。肝脏病理检查结果可见,对照组肝组织正常;暴露组肝细胞水样变性,脂肪样变性,气球样变性,肝细胞点状、灶状、碎片状坏死,汇管区可见炎细胞浸润,少许纤维增生。结论MT表达的降低可能是亚砷酸钠肝毒性的一个易感因素。
Objective To investigate the effect of MT-1 and MT-2 in liver damage in chronic inorganic arsenate exposure mice and to explore the mechanism of arsenic-induced liver damage. Methods The male Kunming mice were randomly divided into control and exposed groups. The control group was given ordinary feed and tap water. The exposed group was given ordinary feed and 300 mg/L of sodium arsenite solution by drinking water. After 10 months of treatment, the activity of serum alanine aminotransferase (ALT), aspartate aminotransferase(AST) and globulin (Glb) content were determined. The total RNA was extracted by the TRIzol-phenol-chlorofor-method from the liver tissue. The quantity of the RNA was determined by spectrophotometry and its purity was judged at a ratio of A260/A280. Then real-time PCR(RT-PCR) was used to measure the mRNA expression of MT-1 and MT-2. Results Compared with the control group, serum ALT, AST activity and Glb content were higher and the MT-1 and MT-2 mRNA content was lower in the exposed group (P<0.05). Conclusion Maybe the down-regulation of MT expression is a susceptible factor in trivalent arsenate incuced hepatotoxicity.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2007年第10期755-758, 图版1,共5页
Journal of Environment and Health
基金
国家自然科学基金资助项目(30471592)
科技部国际科技合作与交流专项经费资助项目[2005DFA30640(A类)]
关键词
砷
金属硫蛋白
基因表达
Arsenic
Metallothionein
Gene expression
