饮用水有机提取物致大鼠肝脏损伤中GSTs活性及其编码基因表达的变化  

Variations in GSTs Activities and Related Gene Expressions of Rats with Liver Injury Caused by Organic Extracts from Drinking Water

作　　者：杨光红[1] 桂晓玲[1] 岑延利[1] 张爱华[1] 敖云霞 李骞[1] 王士然[1] 

机构地区：[1]贵阳医学院公共卫生学院,贵州550004

出　　处：《环境与职业医学》2015年第3期206-210,共5页Journal of Environmental and Occupational Medicine

基　　金：贵州省科技厅资助项目(编号:黔科合J字[2012]2041号);贵阳市科学技术计划项目(编号:筑科合同[2013103]19号)

摘　　要：[目的]观察饮用水有机提取物对大鼠肝脏谷胱甘肽-S-转移酶(glutathione-S-transferase,GSTs)活性及谷胱甘肽-S-转移酶A1(GSTA1)基因m RNA和蛋白表达的影响,探讨其在饮用水有机提取物肝脏损伤中作用。[方法]采用固相萃取法提取水样中有机污染物,50只SD大鼠随机分成5组,分别为空白对照组、溶剂对照组(玉米油)和低、中、高3个染毒组(剂量分别为每天5、20、80 L/kg·bw),进行经口灌胃染毒12周。分光光度法检测GSTs的活性,实时荧光定量聚合酶链反应法和Western blot法分别检测GSTA1基因的m RNA和蛋白质表达水平,同时检测肝功能各指标。[结果](1)大鼠肝脏GSTs酶活性:与空白对照、溶剂对照及低剂量组相比,中剂量[(50.66±5.62)U/mg蛋白]和高剂量组[(39.80±12.95)U/mg蛋白]的GSTs酶活性升高(P<0.05);与中剂量组相比,高剂量组GSTs的酶活性则明显降低(P<0.05)。(2)GSTA1的m RNA及蛋白表达水平:中、高剂量组高于空白对照组、溶剂对照及低剂量组(P<0.05);而与中剂量组相比,高剂量组GSTA1的m RNA表达水平下降(P<0.05)。Western blot检测结果显示,随染毒剂量的增加,GSTA1蛋白表达呈先升高后降低的趋势,与空白对照、溶剂对照及低剂量组比较,中、高剂量组的升高,差异具有统计学意义(P<0.05);而与中剂量组比较,高剂量GSTA1的蛋白表达则下降(P<0.05)。(3)肝功能指标:与对照组比较,血清胆碱酯酶(CHE)在中、高剂量染毒组升高,差异均具有统计学意义(P<0.05);丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转换酶(AST)则仅在高剂量组升高(P<0.05)。总蛋白(TP)和白蛋白(ALB)在高剂量组下降(P<0.05)。GSTA1蛋白表达、GSTs活性与染毒大鼠肝脏CHE水平均呈正相关关系(r=0.490 5,r=0.685 2;P<0.05)。[结论]在本实验条件下,饮用水有机提取物较高剂量染毒可上调大鼠肝脏GSTA1的m RNA及蛋白质表达水平,调控GSTs活性改变,从而导致大鼠肝细胞对毒物的易感�[ Objective ] To observe glutathione-S-transferase （GSTs） enzyme activities and the mRNA and protein expressions of GSTA1 gene in the liver of rats exposed to organic extracts （OEs） from drinking water, and to assess the effect of organic extracts from drinking water in the liver injury. [ Methods ] Organic extracts in drinking water were extracted by solidphase extraction method. Fifty SD rats were randomly divided into blank control group, solvent control group （corn oil）, and three OEs exposed groups （5, 20, and 80L/kg body weight per day）. The exposure via gavage lasted 12 weeks. Spectrophotometry was used to measure the enzyme activities of GSTs. The mRNA and protein levels of GSTA1 gene were measured by real-time quantitative and Western blot method separately. Liver function was also detected. [ Results ] （1） Compared with the blank control, solvent control, and low dose groups, the enzyme activities of GSTs in the middle dose group [（50.66 ± 5.62）U/mg protein] and the high dose group [（39.80± 12.95）U/mg protein] both obviously increased （P 〈 0.05）. While compared with the middle dose group, the activities of GSTs in the high dose group significantly decreased （P 〈 0.05）. （2） The mRNA and protein expression levels of GSTA1 in the middle and high dose groups were significantly higher than those in the blank control, solvent control, and low dose groups （P〈0.05）. While compared with the middle dose group, the mRNA expression level of GSTAI in the high dose group significantly decreased （P 〈0.05）. The results of Western blot showed that with the incease of exposing dose, the expression of GSTA1 protein increased at first then decreased. Compared with the blank control, solvent control, and low dose groups, the GSTA1 protein in the middle and high dose groups elevated significantly （P〈 0.05）. While compared with the middle dose group, the GSTA1 protein in the high dose group significantly decreased （P 〈 0.05）. （3） Compared with the contr

关 键 词：饮用水 有机提取物 谷胱甘肽-S-转移酶 GSTA1 肝脏毒性 

分 类 号：R114[医药卫生—卫生毒理学]