乙苯对大鼠尿中苯乙醇酸、苯乙醛酸水平、肾组织超微结构及线粒体凋亡相关蛋白表达的影响  

Influence of ethylbenzene on the levels of mandelic acid and phenylglyoxylic acid in urine, ultrastructure and the expressions of Mitochondrial apoptotic-related proteins in the rat nephridial tissues

作　　者：王延让[1] 杨德一[1] 张明[1] 王倩[2] 刘静[1] 李建国[1] 张昊[1] 杨雪莹[1] 

机构地区：[1]天津市疾病预防控制中心职业病预防控制所,300011 [2]天津市医药科学研究所

出　　处：《中华劳动卫生职业病杂志》2009年第11期655-659,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：天津市自然科学基金重点项目（07JCZDJC08500、09JCZDJC20900）;天津市卫生局科学基金项目（07KZ72）

摘　　要：目的探讨亚慢性染毒乙苯对大鼠尿中苯乙醇酸、苯乙醛酸和肾组织超微结构及线粒体凋亡相关蛋白表达的影响。方法SD大鼠雄性40只，随机分为对照组、低剂量组、中剂量组和高剂量组，每组10只，分别染毒0、433．5、4335．0和6500．0mg／m^3的乙苯，每天6h，每周5d，共13周，建立大鼠乙苯亚慢性染毒模型；高效液相色谱法测定染毒后大鼠尿中苯乙醇酸（MA）和苯乙醛酸（PGA）水平，电镜观察肾组织的超微结构，Western blot法检测肾组织B细胞淋巴瘤-2相关联X蛋白（Bcl-2associated X protein，Bax）、B细胞淋巴瘤-2（B—cell lymphoma／leukemia-2，Bcl-2）、细胞色素C（eytochrome C，Cyte）、细胞凋亡蛋白酶（Caspase-3）、半胱天冬酶-9（Caspase-9）等线粒体凋亡相关蛋白的表达。结果中、高剂量组大鼠尿中MA[（0．303±0．148）、（0．404±0．154）mg／L]和PGA[（0．168±0．104）、（0．174±0．092）mg／L]水平均明显高于对照组和低剂量组，差异有统计学意义（P〈0．05），且MA、PGA均与乙苯外暴露剂量呈现一定的剂量一效应关系（r=0．827，r=0．720，P〈0．05）。高剂量组大鼠肾组织线粒体普遍肿胀，呈空泡状，部分线粒体嵴排列紊乱，线粒体嵴消失。巾、高剂量组大鼠。肾组织Bax表达水平均明显高于对照组，差异有统计学意义（P〈0．05）；各剂量组大鼠肾组织Cyte和Caspase-9表达水平均明显高于对照组，差异有统计学意义（P〈O．05）；中、高剂量组大鼠肾组织Caspase-3表达水平均明显高于对照组和低剂量组，差异有统计学意义（P〈0．05）；与对照组相比，各剂量组大鼠肾组织Bcl-2表达水平均明显下降差异有统计学意义（P〈0．05）。结论乙苯致大鼠肾组织细胞线粒体损伤可能是乙苯细胞毒作用机制之一，其机制可能与线粒体凋亡相关蛋白表达有关，大鼠尿中苯乙醇酸和苯乙醛酸�Objective To investigate the influnence of ethylbenzene on the levels of mandelic acid （MA） and phenylglyoxylic acid （PGA） in urine, the uhrastructure and the expressions of mitochondrial apoptotic-related genes in the rat nephridial tissues. Methods Four groups of 10 males of Sprague-Dawley rats were allocated randomly into four groups： control （C） group, low （L） gruup, moderate （M） group and high（H） group, and inhaled daily with different doses of ethylbenzene： O, 433.5 mg/m^3, 4335 mg/m^3, and 6500 mg/m^3 6 h per day, 5 days per week for 13 weeks. The mandelic acid and phenylglyoxylic acid in the urine was assayed by high performance liquid chromatography. The uhrastructnre of nephridial tissue was observed via electron microscope. The protein expression levels of Bax, Bcl-2, cytochrome C, Caspase-9 and Caspase-3 in nephridial tissues were measured by Western blot, respectively. Results The levels of MA [（0.303±0.148） mg/L, （0.404± 0.154） mg/L] and PGA [（0.168±0.104） mg/L, （0.174±0.092） mg/L] in the urine of M and H groups were significantly higher than that in the control and L group[（0.08±0.070） mg/L,（0.041±0.029） mg/L]（P〈0.05, respectively）. It has been shown a dose-effect relationship between the contents of MA, PGA and MA＋PGA and inhaled ethylbenzene, respectively. The mitochondria of rat nephridial tissue of H group became a compact and vacuolar structure with disorder and loss of cristae. The expression levels of Bax in mitochondria of nephridial tissues of M and H groups were significantly lower than that in the control group （P〈0.05）. Caspase-3 expression level in H group was remarkably higher than that in the control group（P〈0.05）. Compared with the control group, the expression levels of cytochrome C and Caspase-9 were enhanced, while the expression levels of Bcl-2 were restrained in all ethylbenzene-treated groups （P〈0.05, P〈0.05, respectively）. The expression levels of Caspase-3 in M and H groups were significant

关 键 词：乙苯 大鼠 病理学 

分 类 号：R686[医药卫生—骨科学]