Effects Of Nonylphenol On The Morphology Of Mouse Follicles And Serum Estrogen And Progesterone Levels

Objectives In this study,the effects of nonylphenol on the morphology of ovarian follicles and the levels of estrogen and progesterone in mice were studied by intragastric administration of nonylphenol solution to study the reproductive toxicity of nonylphenol.Methods Thirty female Kunming（KM）mice were randomly divided into control group（corn oil）and low dose group（25 mg/kg）and high dose group（100 mg/kg）,a total of 3groups,10 in each group.The drug was inoculated by a nonylphenol solution,and the dose was 0.1 ml/10 g once a day,5 days per week,and continuously exposed for 12 weeks.1General condition and weight:observe the general daily activities of animals,drinking water diet,stool traits,etc.,changes in skin color,fur smoothness;2 24 hours after the last exposure,the eyeballs were taken and the mice were sacrificed,quickly separate the bilateral ovaries,weigh,calculate the organ coefficient;3 after taking the eyeball to take blood,separate the serum,enzyme-linked immunosorbent assay for serum estradiol（E₂）,progesterone（P）and promote Follicle stimulating hormone（FSH）hormone levels;4Obtain ovarian tissue,observe the pathological changes of mouse ovary and the number of follicles at various levels;5 Preparation of ovarian cell suspensions,detection of ROS levels in ovarian tissue using fluorescent dyes（H₂DCFDA）and flow cytometry;6Preparation of ovarian tissue homogenate,determination of nitric oxide（NO）and nitric oxide synthase（NOS）activity in mouse ovary;7 ovarian tissue,immunohistochemical method for determination of ovarian tissue 8-OHdG Protein expression level;8 Take ovarian tissue,determine the apoptosis of ovarian tissue cells by TUNEL method.Results 1 Compared with the control group,the exposed group had a weak mental state,decreased fur gloss,reduced activity,occasional loose stools,and decreased intake and drinking.2 The weight of the 6-week and 12-week mice in the control group,the low-dose group and high-dose group were（36.56±0.23）g,（33.67±2.76）g,（38.61±2.20）g,（38.54±2.52）g,（38.67±2.76）g,（40.61±2.20）g.the difference was statistically significant（F=11.602,P<0.05;F=11.963,P<0.05）,High dose group mice gain weight.3 The ovarian wet weight and ovarian coefficient of the control group,low dose group and high dose group were（0.29±0.13）g,（0.28±0.09）g,（0.17±0.01）g,（0.84±0.46）%,（0.80±0.36）%,（0.43±0.54）%,the difference was statistically significant（F=369.586,P<0.05;F=244.713,P<0.05）.The ovarian wet weight and ovarian coefficient of the high dose group were significantly decreased.4 The serum levels of E₂,P and FSH in the control group,low-dose group and high-dose group were（17.27±1.89）pg/ml,（15.32±2.27）pg/ml,（13.46±1.62）pg/ml;（2.46±0.39）ng/ml,（2.23±0.25）ng/ml,（1.99±0.23）ng/ml;（4.76±1.62）ng/m l,（3.80±0.68）ng/ml,（2.84±0.87）ng/ml,respectively,the difference was statistically significant(F=7.188,P<0.05;F=6.334,P<0.05 F=7.164,P<0.05,The serum levels of E2,P and FSH in the high dose group were significantly decreased.5 The total number of follicles in the control group,low-dose group and high-dose group were 29.17±1.47,25.17±1.17,18.17±0.75,respectively,and the difference was statistically significant(F=136.098,P<0.05.The number of total follicles in the high dose group was significantly reduced.6 The ROS levels in the ovarian tissues of the control group,the low-dose group and the high-dose group were 70.37±9.85,106.18±18.72,and 209.28±44.41,respectively.The difference was statistically significant（F=19.037,P<0.05）.Ovarian tissue ROS levels are significantly increased.7 The NO activities in the ovarian tissues of the control group,thelow-dosegroupandhigh-dosegroupwere（23.63±0.69）μmol/gpro,（23.04±1.14）μmol/gpro,and（12.04±0.65）μmol/gpro,respectively.The difference was statistically significant（F=348.731,P<0.05）,NO activity in ovarian tissue of mice in high dose group was significantly reduced.8 The activity of NOS in ovarian tissue of control group,low dose group and high dose group were（0.005±0.0003）U/mgpro,（0.0039±0.0004）U/mgpro,（0.0014±0.0014）U/mgpro,respectively.The difference was statistically significant(F=22.739,P<0.05.The activity of NOS in ovarian tissue of mice in high dose group was significantly decreased.9 The mean optical density values of8-OHdG protein expression in ovarian tissue of control group,low dose group and high dose group were 0.0099±0.0004,0.0389±0.0016 and 0.0763±0.0131,respectively.The difference was statistically significant（F=76.214,P<0.05）,high dose group 8-OHdG protein expression level increased.10 The apoptosis rate of ovarian cells in the control group,low-dose group and high-dose group were 5.6%,22.00%,and 44.00%,respectively.The difference was statistically significant（c²=817.267,P<0.05）.The apoptotic rate increased significantly.Conclusions 1 Nonylphenol exposure resulted in weight gain,ovarian wet weight and decreased ovarian coefficient in mice.2 Nonylphenol exposure resulted in a decrease in serum E₂,P and FSH levels in mice.3 The exposure of nonylphenol resulted in a decrease in the total follicles,primary follicles,and mature follicles,and increased number of atresia follicles,high dose of nonylphenol is more effective.4 Nonylphenol exposure resulted in decreased NO and NOS activity in ovarian tissue,increased ROS activity and 8-OHdG protein expression,leading to apoptosis of ovarian tissue,high dose of nonylphenol is more effective.Figure 5;Table 8;Reference 135...