亚砷酸钠染毒对SD大鼠肝脏的损伤作用及血清相关蛋白的表达

Effects of sodium arsenite on liver injury and expression of serum-associated proteins in SD rats

目的建立慢性砷中毒模型,探讨亚砷酸钠对SD大鼠肝脏的损伤作用,同时测定血清相关蛋白含量,为发现早期生物标志物提供依据。方法健康成年SD大鼠112只,雌雄对半,随机分为对照组(A组和E组)、低剂量组(B组和F组)、中剂量组(C组和G组)和高剂量组(D组和H组),共8组,采用自由饮水方式进行染毒,对照组(A、E):0 mg/L,低剂量组(B、F):2 mg/L,中剂量组(C、G):10 mg/L,高剂量组(D、H):50 mg/L,其中A^D组染毒三个月,E^H组染毒6个月,检测大鼠尿砷含量,观察肝脏组织病理学变化,测定血清羧肽酶(CPD)、I型胶原蛋白(COI 1)、大鼠热休克蛋白90α的含量。结果亚砷酸钠染毒3个月和6个月,大鼠肝脏重量基本上呈下降趋势,且在中剂量和高剂量下降较为明显(F=3.246,F=0.989,P<0.05),但肝脏系数变化无统计学意义(F=0.188,F=1.542,P>0.05);随着染毒剂量的增加,肝细胞排列逐渐紊乱,炎性细胞浸润程度增加,肝纤维组织增生,在高剂量组出现肝细胞变性坏死;大鼠血清的羧肽酶(CPD)、热休克蛋白90α(HSP90α)、一型胶原蛋白(COI 1)含量在3个月染毒组差异均无统计学意义(F=1.387,F=0.532,F=1.005,P>0.05),但在6个月染毒组,大鼠羧肽酶(CPD)、一型胶原蛋白(COI 1)差异无统计学意义(F=2.283,F=0.488,P>0.05),热休克蛋白90α(HSP90α)差异具有统计学意义(F=2.086,P<0.05)。结论长期饮水砷暴露可导致肝损伤,热休克蛋白90α在慢性砷中毒中的上调表达提示需要进一步验证热休克蛋白90α是否可以作为早期预警砷中毒的效应指标。

Objective To establish a chronic arsenic poisoning model,explore the liver damage of sodium arsenite on SD rats,and determine the content of serum related proteins in order to provide a basis for the discovery of early biomarkers.Methods A total of 112 healthy adult SD rats,half male and half female were randomly divided into control group(Group A and Group E),low dose group(Group B and Group F),middle dose group(Group C and Group G)and high dose group(Group D and Group H).The rats were divided into 8 groups and exposure to poison by mean of free drinking water:control group(Group A and Group E):0 mg/L,low dose group(Group B and Group F):2 mg/L,middle dose group(Group C and Group G):10 mg/L,high dose group(Group D and Group H):50 mg/L,in which Group A^D was exposed for 3 months,and Group E-H was exposed for 6 months to measure the urinary arsenic content,observe liver histopathological changes,and measure the serum carboxypeptidase(CPD),type I collagen(COI 1)and heat shock protein 90(HSP 90).Results The weight of the rat liver basically showed a downward trend after exposure to sodium arsenite at three and six months,and the decrease was significant at medium and high doses(F=3.246,F=0.989,P<0.05),but the liver coefficient was not statistically significant(F=0.188,F=1.542,P>0.05);With the increase of toxic dose,the arrangement of hepatocytes was gradually disordered,the infiltration degree of inflammatory cells was increased,hepatic fibrosis was proliferated,and hepatocyte degeneration and necrosis occurred in the high-dose group;Carboxypeptidase of rat serum(CPD),heat shock protein 90 alpha(HSP90 alpha),type I collagen(COI)content were observed no statistically significant difference in in the infected group in three months((F=1.387,F=0.532,F=1.005,P>0.05),but in the six months infected group,rats carboxypeptidase(CPD),type I collagen(COI)there was no statistically significant difference((F=2.283,F=0.488,P>0.05).HSP90 showed a statistically significant difference(F=2.086,P<0.05).Conclusion Long-term drinking water arsenic exposure can lead to liver injury.The up-regulation of heat shock protein 90 expression in chronic arsenism suggests that heat shock protein 90 should be used as an early warning indicator of arsenism.