| Objective To observe the histopathological changes of male Wistar rats and determine the expression of inhibin B gene and its protein by intragastric administration of subacute Cyfluthrin for 6 weeks.Objective to explore the effect of Cyfluthrin on testicular structure and INHB in male Wistar rats and the regulation of LBP.Methods According to the random number table method,Wistar rats were divided into control group,Cy low-dose group,Cy middle-dose group,Cy high-dose group,LBP control group,LBP+Cy middle-dose group,10 rats in each group,and the concentrations were rats.Body weight was given 6.25mg/kg(low),12.5mg/kg(medium),25mg/kg(high)Cy and100mg/kg LBP.After continuous gastric administration for 6 weeks,he was put to death by anesthesia.The rat testis tissue was taken to observe the pathological changes of the tissue structure,and the expression levels of INHα and INHβB m RNA and protein in the testis were detected by immunohistochemistry,western blotting test and RT-q PCR method.Results(1)After 6 weeks of intragastric administration of cyfluthrin pesticide,the blood routine test found that the white blood cell count was significantly increased in the low,medium and high dose groups than the control group(P<0.05),and the white blood cell count in the high dose group was higher than that in the medium dose group,Low-dose group(P<0.05);RBC,HGB,PLT and other indicators were not statistically different between groups(all P>0.05).(2)For 6 weeks after exposure,HE staining results showed that the seminiferous tubules of the control group and the LBP control group were clearly demarcated and arranged loosely.Sertoli cells and spermatogenic cells in the lumen were arranged tightly and clearly demarcated without abnormalities;low-dose group The seminiferous tubules of the tissues are clearly demarcated and arranged loosely.The spermatogenic cells and supporting cells are arranged tightly,and some seminiferous tubules are irregular in shape;the middle-dose group has clear boundaries and loosely arranged seminal tubules,with spermatogenic cells in the seminiferous tubules And supporting cells are reduced,a small amount of seminiferous tubules are irregular in shape;in the high-dose group,the seminiferous tubules are clearly demarcated and arranged loosely,a large number of spermatids and supporting cells in the seminiferous tubules are reduced,and a large number of seminiferous tubules are irregular in shape.Spermatogenic cells can be seen in the seminiferous tubules;the seminiferous tubules in the Cy+ medium dose group are clearly demarcated and arranged loosely.The spermatogenic cells and supporting cells in the seminiferous tubules are arranged tightly,and some seminiferous tubules are irregular in shape.(3)After 6 weeks of exposure,the results of rat immunohistochemical experiments showed that the positive expression products of INHα and INHβB protein were brown-yellow.INHα and INHβB proteins were mainly expressed in the cytoplasm of spermatogenic cells at various levels in the testis,while the remaining cells and nuclei were not seen Obviously expressed.The nucleus immunohistochemical staining of spermatogenic cells in each group of rat testis tissues is mainly blue.The brown particles of INHα and INHβB protein in testis tissues of the control group and the Lycium barbarum polysaccharide control group are evenly distributed;With increasing dose,the positive staining distribution of INHα and INHβB was sparser than that of the control group,and the degree of staining expression was lighter.According to statistics,the positive rates of each group of INHα were 93.02636%,93.76027%,87.70575%,78.42155%,61.71080%,85.03282%.Compared with the control group,the difference between the low,medium and high dose group was statistically significant(P<0.05);the difference between the LBP+Cy medium dose group and the LBP control group was statistically significant(P<0.05);the LBP+Cy medium dose group Compared with the Cy middle dose group,the difference was statistically significant(P<0.05).The positive rates of INHβB groups were 78.062531%,79.592911%,40.494335%,25.379118%,15.135840%,48.150013%.Compared with the control group,the differences between the low,medium and high dose groups were statistically significant(P<0.05);LBP+Cy The difference between the middle dose group and the LBP control group was statistically significant(P<0.05);the difference between the LBP+Cy middle dose group and the Cy middle dose group was statistically significant(P<0.05).(4)After 6 weeks of exposure,the results of transmission electron microscopy showed that the control group was similar to the LBP control group.The sperm cell membrane was intact,the organelles were abundant,the nucleus was round,the nuclear membrane was complete and clear,and the chromatin was evenly distributed;the acrosome was continuous and complete,The number of mitochondria is abundant and evenly distributed;the rough endoplasmic reticulum is abundant and evenly distributed.Sperm cells in the low-dose group showed mild edema,cell membranes were intact,nuclei were round,nuclear membranes were intact and clear,and chromatin was evenly distributed;acrosomes were continuous and complete,and mitochondria were slightly swollen.Sperm cells in the medium-dose group were moderately edema,cell membranes were intact,nuclei were irregular,chromatin was sparse,nuclear membrane was locally raised,perinuclear space was widened,acrosome structure was intact;mitochondria were moderately swollen.Sperm cells in the high-dose group showed severe edema,intracellular vacuoles,irregular nuclei,sparse chromatin,blurred nuclear membrane,and blurred acrosome structure;mitochondria were heavily swollen with vesicular cristae,and local intramembrane matrix became weak,Vacuoles become;the rough surface is rich in endoplasm,and there are more lipid droplets.Sperm cells in the Cy+ LBP medium-dose group showed mild edema,cell membranes were intact,nuclei were round,nuclear membranes were intact and clear,and chromatin was evenly distributed;acrosomes were continuous and intact,mitochondria were slightly swollen,and the cristae were in the shape of vesicles and local membrane The matrix becomes lighter;the rough endoplasmic reticulum is abundant and evenly distributed.(5)After 6 weeks of exposure,PCR method was used to detect the expression levels of testosterin INHα and INHβB m RNA.The PCR test results showed that compared with the control group,the low,medium,and high dose groups showed a gradual decrease,and the difference between the groups was statistically significant.Significance(P<0.05).There was no statistically significant difference in gene m RNA expression between the control group and the LBP control group(P>0.05).Compared with the LBP control group,the m RNA expression level of the LBP+medium-dose Cy group decreased,and the difference was statistically significant(P<0.05).The difference between the middle-dose Cy group and the LBP+medium-dose Cy group was statistically significant(P<0.05).(6)After 6 weeks of exposure,the protein expression level test results showed that compared with the control group,testosterin INHα and INHβB showed a gradual decline in the low,medium and high groups,and the difference between the groups was statistically significant(P<0.05).There was no statistically significant difference in protein expression between the control group and the LBP control group(P>0.05).Compared with the LBP control group,the expression level of Cy histone at LBP+medium dose decreased,and the difference was statistically significant(P<0.05).There was a statistically significant difference between the medium dose of LBP+medium dose of Cy group in Cy group(P<0.05).Conclusion Cy exposure caused the destruction of rat testis tissue structure.The m RNA and protein expressions of INHα and INHβB decreased,indicating that Cy has a toxic effect on rat testis tissue.LBP intervention has a certain protective effect on rat testis tissue. |
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