摘要
目的探讨转录因子NF-E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)对氯化镉所致大鼠睾丸和卵巢内质网应激的反馈调节作用。方法将48只SD大鼠随机分为对照组,氯化镉低、中、高剂量组,叔丁基对苯二酚(tert-bu-tylhydroquinone,tBHQ)组,tBHQ+氯化镉低、中、高剂量组,共8组,每组6只。对照组、tBHQ组分别给予腹腔注射0.9%生理盐水、20 mg/kg体重tBHQ溶液;氯化镉低、中、高剂量组分别腹腔注射1.14、2.28、4.56 mg/kg体重氯化镉溶液;tBHQ+氯化镉低、中、高剂量组于氯化镉染毒前120 min先行腹腔注射20 mg/kg体重tBHQ溶液后,分别腹腔注射1.14、2.28、4.56 mg/kg体重氯化镉溶液。染毒48 h后处死动物,取睾丸、卵巢组织,采用RT-PCR和Western blot法测定Bip、PERK、Nrf2 mRNA和蛋白的表达水平。结果与tBHQ组比较,tBHQ+氯化镉中剂量组大鼠睾丸Nrf2 mRNA和蛋白表达明显上调(F值分别为144.47、302.88,P<0.05),tBHQ+氯化镉低、中、高剂量组大鼠卵巢Nrf2 mRNA和蛋白表达均明显上调(F值分别为55.97、80.08,P<0.05);与同剂量氯化镉组比较,tBHQ+氯化镉中剂量组大鼠睾丸Nrf2蛋白表达明显上调(F=74.66,P值<0.05),此剂量组大鼠睾丸Bip、PERK表达水平无显著变化;tBHQ+氯化镉高剂量组大鼠卵巢Nrf2蛋白表达明显上调(F=30.58,P<0.05),大鼠卵巢Bip mRNA和蛋白、PERK mRNA表达均出现不同程度的下调(F值分别为33.13、25.28、30.24,P<0.05),且大鼠卵巢Nrf2蛋白表达与Bip mRNA、PERK mRNA表达均呈负相关关系(r值分别为-0.783、-0.817,P<0.05),tBHQ+氯化镉高剂量组大鼠睾丸PERK、Nrf2 mRNA和Bip、PERK、Nrf2蛋白的表达均明显低于氯化镉高剂量组(F值分别为19.73、53.63、25.28、99.98、74.66,P<0.05)。结论在不同氯化镉染毒剂量条件下,tBHQ的干预可不同程度地诱导大鼠睾丸和卵巢的Nrf2表达;大鼠卵巢Nrf2表达的上调,可通过PERK/Nrf2途径减轻ERS水平,以改善镉对卵巢组织的损害。
Objective To investigate the feedback regulatory effects of nuclear factor erythroid 2-related factor 2(Nrf2)on cad-mium chloride(CdCl2)-induced endoplasmic reticulum stress in the testicles and ovaries of rats.Methods Forty-eight Sprague-Daw-ley rats were randomly divided into eight groups(six rats in each group)according to the cadmium chloride exposure dose and whether to administer tert-butylhydroquinone(tBHQ):control group(intraperitoneal injection of 0.9%normal saline);low-dose,medium-dose,and high-dose CdCl2 groups(intraperitoneal injected of CdCl2 solution at 1.14,2.28,and 4.56 mg/kg,respectively);tBHQ group(intraperitoneal injection of 20 mg/kg tBHQ solution);and tBHQ+low-dose,tBHQ+medium-dose,and tBHQ+high-dose CdCl2 groups(pretreatment with 20 mg/kg tBHQ solution 120 minutes before intraperitoneal injection of 1.14,2.28,and 4.56 mg/kg CdCl2 solution,respectively).After 48 hours of exposure,the rats were sacrificed to collect the testicles and ovaries,and the mRNA and protein expression levels of 78 kDa glucose-regulated protein(Bip),PKR-like endoplasmic reticulum kinase(PERK),Nrf2 was de-termined by RT-PCR and Western blot.Results Compared with tBHQ group,the expression of Nrf2 mRNA and protein in the testi-cles in the tBHQ+medium-dose CdCl2 group was significantly increased(F were 144.47 and 302.88,respectively,P<0.05);the ex-pression of Nrf2 mRNA and protein in the ovaries in the tBHQ+low-dose,medium-dose,and high-dose CdCl2 groups was all signifi-cantly increased(F were 55.97 and 80.08,respectively,P<0.05).Compared with the same dose of CdCl2 group,the tBHQ+medi-um-dose CdCl2 group showed significantly increased expression of Nrf2 protein in the testicles(F=74.66,P<0.05),with no signifi-cant differences in the expression of Bip and PERK in the testicles;the tBHQ+high-dose CdCl2 group showed a significant increase in the expression of Nrf2 protein in the ovaries(F=30.58,P<0.05)and varying degrees of reductions in the expression of Bip mRNA and protein(F were 33.13,25.28,and 30.24,respectively,P<0.05)
作者
陈智健
倪秀贤
吴家韵
蔡日东
余日安
CHEN Zhi-jian;NI Xiu-xian;WU Jia-yun;CAI Ri-dong;YU Ri-an(Shenzhen Maternity and Child Healthcare Hospital,Shenzhen 518028,China;School of Public Health,Guangdong Pharmaceutical University;Fuyong Institute for Prevention and Health Care,Bao′an District,Shenzhen)
出处
《环境卫生学杂志》
2024年第1期29-35,共7页
JOURNAL OF ENVIRONMENTAL HYGIENE
基金
深圳市科技计划项目(JCYJ20180301170050533)
深圳市宝安区科技创新局2017年宝安医疗卫生基础研究项目(2017JD064)。
