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

发布时间：2018-05-11 00:09

本文选题：饮用水 + 有机提取物　；参考：《环境与职业医学》2015年03期

【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(P0.05);与中剂量组相比,高剂量组GSTs的酶活性则明显降低(P0.05)。(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(P0.05);而与中剂量组相比,高剂量组GSTA1的m RNA表达水平下降(P0.05)。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(P0.05);而与中剂量组比较,高剂量GSTA1的蛋白表达则下降(P0.05)。(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(P0.05);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(P0.05)。总蛋白(TP)和白蛋白(ALB)在高剂量组下降(P0.05)。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;P0.05)。[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性增强,肝损伤加重。
[Abstract]:[Objective] to observe the effects of organic extracts from drinking water on the activity of glutathione -S- transferase (glutathione-S-transferase, GSTs) and the expression of M RNA and protein of glutathione -S- transferase A1 (GSTA1) gene in rat liver, and to explore the use of solid phase extraction in the extraction of organic pollutants in water samples by the method of solid phase extraction. 50 SD rats were randomly divided into 5 groups: blank control group, solvent control group (corn oil) and low, middle and high 3 infected groups (5,20,80 L/kg / BW per day respectively) for 12 weeks by oral gavage. The activity of GSTs was detected by spectrophotometric method, GSTA1 base was detected by real-time quantitative polymerase chain reaction and Western blot method, respectively. The expression level of M RNA and protein, and the detection of liver function. [results] (1) the activity of GSTs enzyme in the liver of rats: compared with the blank control, the medium dose [(50.66 + 5.62) U/mg protein] and high dose group [(39.80 + 12.95) U/mg protein] increased GSTs activity compared with the blank control group (P0.05), and the high dose group GSTs compared with the medium dose group. The activity of the enzyme decreased significantly (P0.05). (2) the m RNA and protein expression level of GSTA1 were higher in the high dose group than in the blank control group, the solvent control and the low dose group (P0.05), and the m RNA expression level of GSTA1 in the high dose group decreased (P0.05).Western blot detection results showed that the expression of the GSTA1 protein expressed with the increase of the dose. The trend of first increasing and then decreasing, compared with the blank control, compared with the solvent control and low dose group, the high dose group increased, the difference was statistically significant (P0.05), but compared with the middle dose group, the protein expression of high dose GSTA1 decreased (P0.05). (3) the liver function index: compared with the control group, the serum cholinesterase (CHE) was in the high dose group. The difference was statistically significant (P0.05); alanine aminotransferase (ALT), aspartic aminotransferase (AST) increased only in high dose group (P0.05). Total protein (TP) and albumin (ALB) decreased (P0.05).GSTA1 protein expression in high dose group, GSTs activity was positively correlated with CHE level of rat liver (r=0.490 5, r=0.685 2). 0.05) [Conclusion] [Conclusion] in this experiment, the high dose of organic extracts from drinking water can increase the level of M RNA and protein expression of rat liver GSTA1, regulate the changes of GSTs activity, and lead to the enhancement of the susceptibility to poison in rat liver cells and the aggravation of liver injury.

【作者单位】：贵阳医学院公共卫生学院;
【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)
【分类号】：R114

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