Reproductive Toxicity Of Dimethyl Phthalate To C57 Mice

Objective:Dimethyl phthalate is widely used in the world,and is often used in cosmetics,perfume and other care products,mosquito repellent oil,varnish and packaging bags,plastic film and other plastic products.DMP can be detected in water,soil,and the atmosphere,and it enters the organism through the atmosphere,water,food,etc.,posing a threat to the health of the body.At present,there is no research on the reproductive toxicity of DMP in mice at home and abroad.In this experiment,DMP was continuously administered to C57 mice,and the changes of ovarian and testis were observed at 20 and 40 days respectively.The role of mice reproductive toxicity in order to provide information for the toxicological detection and prevention of DMP,and provide a basis for its safe application.Method:80 health C57 mice are randomized into control group（corn oil）and 0.5、1.0、2.0 g·kg^-11 DMP group.Each group holds 20 mice,half male and half female,which have been continuously Intragastric once a day.20 and 40 days after exposure,then we taking blood from the orbital sinus,the mice were killed by dislocation.Taking testicles of male mice and observing serum sex hormone levels（T,FSH,LH）,testicular tissue marker enzyme activity（LDH,SDH,G-6-PD）,ATPase,lipid peroxidation damage（SOD,CAT,GSH-Px,MDA）,sperm count and deformity rate,testicular cell cycle and apoptotic rate.Taking ovary of female mice and observing changes in serum sex hormone levels（P,E₂,FSH,LH）and ovarian cell apoptosis rate.Result:1.Changes in body weight,anogenital distance,testicular organ coefficient and ovarian organ coefficient in each groupMale mice were exposed to poisoning for 20 days.The testicular organ coefficient of 1.0 g·kg^-1DMP group was significantly higher than that of the control group（P<0.05）.The testicular organ coefficient of 2.0 g·kg^-1DMP group was significantly lower than 0.5 and 1.0 g·kg^-1DMP group（P<0.05）.Exposure for 40 days,1.0 and 2.0 g·kg^-1DMP anogenital distance mice was significantly lower than the control group and 0.5 g·kg^-1DMP group（P<0.05）.The testicular organ coefficient of each group was significantly lower than that of the control group（P<0.05）;the testicular organ coefficient of 1.0 and 2.0 g·kg^-1DMP group was significantly lower than that of 0.5 g·kg^-1DMP group（P<0.05）.The ovarian organ coefficient of each group was significantly higher than that of the control group（P<0.05）.The ovarian organ coefficient of the 1.0 g·kg^-1DMP group was significantly lower than that of the 0.5 g·kg^-1DMP group（P<0.05）.The ovarian organ coefficient of the 2.0 g·kg^-1DMP group was significantly higher than that of the0.5 and 1.0 g·kg^-1DMP group（P<0.05）.After 40 days of exposure,the ovarian organ coefficient of 0.5g·kg^-1DMP group was significantly higher than that of the control group（P<0.05）;the ovarian organ coefficient of 1.0 and 2.0 g·kg^-1DMP group was significantly lower than that of the control group and 0.5 g·kg^-1DMP group（P<0.05）;ovarian organ coefficient in 2.0 g·kg^-1DMP group was significantly lower than 1.0g·kg^-1DMP group（P<0.05）.2.Changes in serum levels of sex hormones in mice of each groupMale mice were exposed for 20 days,and the T content in the 0.5 g·kg^-1DMP group was significantly higher than that in the control group（P<0.05）.The T content in the 1.0 and 2.0 g·kg^-1DMP group was significantly lower than that in the 0.5 g·kg^-1DMP group（P<0.05）.The FSH content in the 2.0 g·kg^-1DMP group was significantly higher than that in the control group and 1.0 g·kg^-11 DMP group（P<0.05）.The LH content of each exposed group was lower than that of the control group（P<0.05）.After 40 days of exposure,the T content of each group was significantly lower than that of the control group（P<0.05）.The T content of 1.0 and 2.0 g·kg^-1DMP group was significantly higher than that of 0.5 g·kg^-1DMP group（P<0.05）.The FSH content of each group was lower than that of the control group（P<0.05）.The FSH content of 2.0g·kg^-1DMP group was significantly higher than that of 0.5 g·kg^-1DMP group（P<0.05）.After 40 days of exposure to female mice,the serum E₂ content of the mice in each group was significantly higher than that of the control group（P<0.05）.The serum E₂ content of the mice in the 1.0 and 2.0 g·kg^-1DMP group was significantly lower than 0.5 g·kg^-1DMP group（P<0.05）.FSH content of 2.0 g·kg^-1DMP group was significantly lower than other groups（P<0.05）.The LH content of the mice in the 2.0g·kg^-1DMP group was significantly higher than that in the other groups（P<0.05）.3.Changes in enzyme activity of testis tissue in each group of mice20 days after exposure,the LDH content of each exposure group was higher than that of the control group（P<0.05）.The LDH levels of 1 and 2.0 g·kg^-1DMP group were higher than 0.5 g·kg^-1DMP group（P<0.05）.The content of G-6-PD in the 0.5and 2.0 g·kg^-1DMP group was significantly lower than that in the control group（P<0.05）.The content of G-6-PD in the 1.0 g·kg^-1DMP group was significantly higher than that in the 0.5 g·kg^-1DMP group（P<0.05）;G-6-PD content in 2.0 g·kg^-1DMP group was significantly lower than that in 1.0 g·kg^-11 DMP group（P<0.05）.The SDH content in the 0.5 g·kg^-1DMP group was significantly lower than that in the control group（P<0.05）.The levels in the 1.0 and 2.0 g·kg^-1DMP group were significantly higher than those in the control group and 0.5 g·kg^-11 DMP group（P<0.05）.The LDH content in the 1.0 and 2.0 g·kg^-1DMP groups was significantly lower than that in the control group and 0.5 g·kg^-1DMP group（P<0.05）.The LDH content in the 2.0 g·kg^-1DMP group was significantly lower than 1.0 g·kg^-1DMP group（P<0.05）.The content of G-6-PD in testis homogenate of each group was significantly higher than that of the control group（P<0.05）;the content of G-6-PD in group 1 and2.0 g·kg^-1DMP was significantly higher than 0.5 g·kg^-1DMP group（P<0.05）.The SDH content in the 2.0 g·kg^-1DMP group was significantly higher than that in the control group and 1.0 g·kg^-11 DMP group（P<0.05）.4.Changes of Ca²⁺-Mg²⁺-ATPase and Na⁺-K⁺-ATPase activities in testis of male miceNa⁺-K⁺-ATPase activity in the 1.0 and 2.0 g·kg^-1DMP group was significantly lower than that in the control group and 0.5g·kg^-1DMP group（P<0.05）.20 days after exposure,the Na⁺-K⁺-ATPase activity in the 2.0 g·kg^-1DMP group was significantly lower than 1.0 g·kg^-1DMP group（P<0.05）.The Ca²⁺-Mg²⁺-ATPase activity in each exposure group was significantly lower than that in the control group（P<0.05）.The Ca²⁺-Mg²⁺-ATPase activity in the 2.0 g·kg^-1DMP group was significantly lower than that in the 0.5 and 1.0 g·kg^-1DMP groups（P<0.05）.40 days after exposure,0.5 and 1.0 g·kg^-1DMP group of active Na⁺-K⁺-ATPase were significantly lower（P<0.05）;2.0 g·kg^-1DMP group of active Na⁺-K⁺-ATPase was significantly higher than 0.5 and 1.0 g·kg^-1DMP group（P<0.05）.The activity of Na⁺-K⁺-ATPase in the 1.0 g·kg^-1DMP group was significantly lower than that in the0.5g·kg^-1DMP group（P<0.05）.The Ca²⁺-Mg²⁺-ATPase activity in the 1.0 g·kg^-1DMP group was significantly higher than that in the 0.5g·kg^-1DMP group and the control group（P<0.05）.The Ca²⁺-Mg²⁺-ATPase activity in the 2.0 g·kg^-1DMP group was significantly lower than 1.0 g·kg^-1DMP group（P<0.05）.5.Changes in lipid peroxidation levels in testis of male miceAfter 20 days of exposure,the MDA content in the testis homogenate of each exposure group was significantly lower than that of the control group（P<0.05）;the content of 2.0 g·kg^-1DMP group was significantly lower than 0.5 and 1.0 g·kg^-1DMP group（P<0.05）.The GSH-Px content of the mice in each exposure group was significantly higher than that in the control group（P<0.05）.The SOD levels in the 1and 2.0 g·kg^-1DMP groups were significantly higher than those in the control group and 0.5 g·kg^-11 DMP group（P<0.05）.The CAT content of 0.5 and 1.0 g·kg^-1DMP group was significantly higher than that of the control group（P<0.05）;the CAT content of 1.0 g·kg^-1DMP group was significantly higher than that of 0.5 g·kg^-1DMP group（P<0.05）;the CAT content of the 2.0 g·kg^-1DMP group was significantly lower than that of the other groups（P<0.05）.MDA content in the 0.5 and 1.0 g·kg^-1DMP groups was significantly higher than that in the control group（P<0.05）.The MDA content in the 2.0 g·kg^-1DMP group was significantly lower than that in the other groups（P<0.05）.The GSH-Px content of the mice in each exposure group was significantly lower than that in the control group（P<0.05）.The GSH-Px content in the 1.0 g·kg^-1DMP group was significantly lower than that in the 0.5 g·kg^-1DMP group（P<0.05）.The GSH-Px content in 2.0g·kg^-1DMP group was significantly higher than that in 0.5 and 1.0 g·kg^-11 DMP group（P<0.05）.6.Changes of apoptosis and cycle and ovarian cell apoptosis in testicular cells of mice in each groupMale mice were exposed for 20 days,and the G0/G1 phase of 0.5 and 1.0 g·kg^-1DMP group was significantly higher than that of the control group（P<0.05）.The S phase of each exposed group was significantly lower than that of the control group（P<0.05）;the S phase of 2.0 g·kg^-1DMP group was significantly higher than that of0.5 g·kg^-1DMP group（P<0.05）.The G2/M phase of each group was significantly higher than that of the control group（P<0.05）.The G2/M phase of 1.0 and 2.0g·kg^-1DMP group was significantly lower than that of 0.5 g·kg^-1DMP group（P<0.05）.The apoptosis rate of each group was significantly higher than that of the control group（P<0.05）.After 40 days of exposure,the G0/G1 phase of 0.5 g·kg^-1DMP group was significantly higher than that of the control group（P<0.05）;the G0/G1 phase of1.0 and 2.0 g·kg^-1DMP group was significantly lower than 0.5 g·kg^-1DMP group.（P<0.05）;G0/G1 phase of 2.0 g·kg^-1DMP group was significantly lower than that of control group and 1.0 g·kg^-1DMP group（P<0.05）.The S phase of each group was significantly lower than that of the control group（P<0.05）.The S phase of 1.0 and 2.0g·kg^-1DMP group was significantly higher than that of 0.5 g·kg^-1DMP group（P<0.05）.The G2/M phase of 1.0 and 2.0 g·kg^-1DMP group was significantly higher than that of the control group and 0.5 g·kg^-11 DMP group（P<0.05）.The apoptosis rate of each group was significantly higher than that of the control group（P<0.05）.The apoptosis rate of 1.0 and 2.0 g·kg^-1DMP group was significantly higher than that of 0.5g·kg^-1DMP group（P<0.05）;2.0 g·kg^-1DMP group was significantly lower than the 1.0g·kg^-11 DMP group（P<0.05）.The apoptosis rate of ovarian cells in the 0.5g·kg^-1DMP group was significantly lower than that in the control group（P<0.05）.The apoptosis rate of ovarian cells in the 1.0 and 2.0 g·kg^-1DMP group was significantly higher than that in the control group and 0.5g·kg^-1DMP group（P<0.05）.After 40 days of exposure,the apoptosis rate of ovarian cells in the 0.5g·kg^-1DMP group was significantly lower than that in the control group（P<0.05）.The apoptosis rate of ovarian cells in the 1.0 and 2.0g·kg^-1DMP group was significantly higher than that in the control group and 0.5g·kg^-1DMP group（P<0.05）.7.Changes in sperm count and sperm deformity rate in male mice of each groupAfter 20 days of exposure,the relative sperm count of each exposure group was significantly lower than that of the control group（P<0.05）;the relative sperm count of1.0 g·kg^-1DMP group was significantly higher than that of 0.5 g·kg^-1DMP group（P<0.05）;the relative sperm count in the 2.0 g·kg^-1DMP group was significantly lower than that in the 0.5 and 1.0 g·kg^-1DMP groups（P<0.05）.The sperm deformity rate was significantly higher in the 1.0 and 2.0 g·kg^-1DMP group than in the control group and 0.5 g·kg^-1DMP group（P<0.05）.The sperm abnormality rate in the 2.0g·kg^-1DMP group was significantly higher than 1.0 g·kg^-1DMP group（P<0.05）.After 40 days of exposure,the relative sperm count of 0.5 g·kg^-1DMP group was significantly higher than that of the control group（P<0.05）;the relative sperm count of 1.0 and 2.0 g·kg^-1DMP group was significantly lower than that of the control group and 0.5 g·kg^-1DMP.The relative sperm count of the 2.0 g·kg^-1DMP group was significantly higher than that of the 1.0 g·kg^-1DMP group（P<0.05）.The sperm deformity rate of each exposure group was significantly higher than that of the control group（P<0.05）.The sperm deformity rate of 1.0 and 2.0 g·kg^-1DMP group was significantly higher than that of 0.5 g·kg^-1DMP group（P<0.05）.Conclusion:1.DMP can cause decreased serum LH and T content and increased FSH activity in male mice;E₂ and LH levels in serum and FSH activity decreased in female mice.2.With the prolonged exposure time,DMP can lead to the increase of LDH content in mouse testis tissue first and then decrease;the change of G-6-PD content first decreases and then increases;the high dose DMP leads to the increase of SDH content.3.DMP can reduce the activity of Na⁺-K⁺-ATPase in mouse testis tissue.After 40days of exposure,the activity of Ca²⁺-Mg²⁺-ATPase in testis tissue of 1.0 g·kg^-1DMP group increased.4.After 20 days of exposure,MDA content in mouse testis homogenate decreased;SOD content increased;CAT content increased in 0.5 and 1.0 g·kg^-11 DMP group,and CAT content decreased in 2.0 g·kg^-1DMP group.After 40 days of exposure,MDA content increased in 0.5 and 1.0 g·kg^-1DMP groups,and MDA content decreased in 2.0 g·kg^-11 DMP group.As the exposure time is prolonged,DMP can cause the content of GSH-Px to increase first and then decrease.5.DMP affects the cell cycle of mouse testis and induces apoptosis of testicular and ovarian cells to some extent.6.DMP causes a decrease in the number of sperm in mice and an increase in sperm abnormality in mice.