摘要
目的探究细胞外调节蛋白激酶(extracellular regulated protein kinase,ERK)通路活化参与邻苯二甲酸二丁酯(dibutyl phthalate,DBP)诱导昆明(KM)小鼠睾丸损伤的作用机制。方法性成熟雄性KM小鼠56只,随机分为8组:对照组、50mg/(kg·d) DBP组、50 mg/(kg·d)维生素E(vitmin E,VE)组、2 mg/(kg·d)尼莫地平(nimodipine,Ni)组、DBP+VE组、DBP+Ni组、Ni+VE组、DBP+Ni+VE组,处理28 d后测定小鼠体重、睾丸重量、脏器系数及精子密度,观察睾丸的组织病理学结果,DCFH-DA荧光定量法检测小鼠睾丸中的活性氧(reactive oxygen species,ROS)、硫代巴比妥酸(TBA)比色法检测丙二醛(malondialdehyde,MDA)含量,酶联免疫吸附法(ELISA)检测钙调蛋白(calmodulin,CaM)含量及磷酸化ERK(p-ERK)水平。结果与对照组比较,50 mg/(kg·d) DBP组小鼠体重、睾丸重量以及睾丸脏器系数下降为(36. 48±0. 99) g、(0. 25±0. 01) g、(0. 54±0. 09)%(P <0. 05),精子密度下降为(11. 70±0. 23)×10~6/m L(P <0. 05),睾丸组织损伤程度增加,ROS荧光强度、MDA含量增加为(1698. 18±77. 58)、(1. 65±0. 13)μmol/g prot(P <0. 05),CaM含量下降为(45. 61±2. 69)μg/m L(P <0. 05),p-ERK水平增加为(1150. 43±48. 79) pg/m L(P <0. 05);加入抗氧化剂VE和钙离子通道拮抗剂Ni处理后,与50 mg/(kg·d)DBP组比较,DBP+Ni+VE组小鼠体重、睾丸脏器系数增加为(40. 69±0. 75) g、(0. 69±0. 03)%(P <0. 05),精子密度增加为(13. 50±0. 16)×10~6/m L(P <0. 05),睾丸组织损伤程度缓解,ROS荧光强度、MDA含量降低为(1080. 60±98. 64)、(1. 06±0. 13)μmol/g prot(P <0. 05),CaM含量增加为(54. 76±1. 74)μg/m L(P <0. 05),p-ERK水平下降为(904. 55±64. 73) pg/m L(P <0. 05)。结论 VE作为抗氧化剂,Ni作为钙离子通道拮抗剂,均可不同程度地降低DBP对小鼠睾丸组织的损伤,推测DBP可能经氧化应激与Ca2+信号激活ERK1/2通路,从而导致小鼠睾丸组织的损伤。
Objective To investigate the role of extracellular regulated protein kinase (ERK)pathway activation in the testicular injury induced by dibutyl phthalate (DBP)in KM mice.Methods A total of fifty-six male KM mice were randomly divided into 8 groups;control group,50mg/(kg·d)DBP group,50mg/(kg·d)vitamin E (VE) group,2mg/(kg-d)nimodipine (Ni)group,DBP +VE group,DBP +Ni group,Ni + VE group and DBP +Ni +VE group.After consecutive 28days of treatment,the body weight,testis weight,organ coefficient and sperm density of mice were measured.The laistomorphological damage of testis was observed by light microscope.The contents of reactive oxygen species (ROS)and malondialdehyde (MDA)in testicular homogenate of mice in each group were detected by DCFH-DA fluorescence and thiobarbituric acid (TBA)colorimetric method,respectively.The contents of calmodulin (CAM)and level.of phosphorylated ERK (p-ERK)were measured by enzyme-linked immunosorbent assay (ELISA).Results Compared with control group,the body weight,testis weight and testicular organ coefficient of mice in 50mg/(kg·d)DBP group decreased to (36.48± 0.99)g,(0.25±0.01)g,(0.54±0.09)%(P<0.05),sperm density decreased to (11.70±0.23)× 10^6/mL (P <0.05),and the degree of testicular tissue injury increased with the fluorescence intensity of ROS and the content of MDA increased to (1698.18±77.58),(1.65±0.13)μmol/g prot (P <0.05)respectively.Meanwhile the content of CaM decreased to (45.61±2.69)μg/mL (P <0.05)as well as the level of p-ERK increased to (1150.43±48.79)pg/mL (P <0.05).After adding VE as antioxidant and Ni as a calcium channel antagonist,compared with 50mg/(kg·d)DBP group,the body weight and testicular organ coefficient of mice in DBP +Ni +VE group increased to (40.69±0.75)g,(0.69±0.03)%(P <0.05),sperm density increased to (13.50±0.16)× 10^6/mL (P <0.05),and the degree of testicular tissue injury decreased with the fluorescence intensity of ROS and the content of MDA decreased to (1080.60±98.64),(1.06±0.13)μmol/g prot (P <0.05)respectively.Meanw
作者
李娟
吴喆
程建
Li Juan;Wu Zhe;Cheng Jian(Department of Internal Medicine,College of Clinical Medicine,Hubei University of Science and Technology,Xianning 437100,China)
出处
《卫生研究》
CAS
CSCD
北大核心
2018年第6期956-962,共7页
Journal of Hygiene Research
