Damage Of Olaquindox To Male Mice’s Reproductive System And VE,APS Intervention On Its Toxic Effects

Olaquindox(OLA)has good antibacterial and growth-promoting effects,and is often misused by some producers driven by economic interests.In recent years,olaquindox(Olaquindox,OLA)has occasional occurrences of olaquindox poisoning.Numerous studies have shown that olaquindox can cause oxidative damage to the body and can also induce apoptosis of spermatogenic cells.This study hypothesized that olaquindox had oxidative damage on the testes of mice.Different doses of olaquindox poisoning models and intervention groups of vitamin E(VE)and astragalus polysaccharides(APS)were established to study the effects of olaquindox on the reproductive function of male mice.And VE,APS intervention of its toxic effects.80 male mice of about 25 g were randomly divided into blank control group,negative control group(OLA 0mg/kg),low OLA group(OLA 80mg/kg),middle OLA group(OLA 120mg/kg),High OLA group(OLA 240mg/kg),APS group(OLA 240 mg/kg+APS 20mg/kg),VE group(OLA 240mg/kg+VE 15mg/kg),VE+APS group(OLA 240mg/kg +VE 15mg/kg +APS 20mg/kg),10 mice in each group were continuously fed for 35 days,and 24 hours after the end of the exposure,mice were killed by cervical dislocation.The experiments were divided into olaquindox and VE,APS intervention on the growth and spermatogenesis of mice,the impact of testicular and epididymal histomorphology,the testosterone on the antioxidant function and testosterone synthesis in four parts.1.Effect of olaquindox,VE and APS intervention on growth and semen quality of miceTests were conducted to determine the effects of olaquindox,VE,and APS interventions on the growth and spermatogenesis of mice by measuring the end-ofperiod test body weight,testis coefficients,seminal gonads,and sperm parameters(sperm abnormalities,vitality,viability and density).influences.Test results show:(1)There was no significant difference between the OLA group,the low OLA group,the blank control group,and the negative control group in the body weight of male mice at the end of the experiment period.The high OLA group was significantly lower than the low OLA group(P < 0.05).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.(2)Low testis coefficient There was no significant difference between the OLA group and the control group.The OLA group and the high OLA group were significantly lower than the blank control group(P<0.05).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.The seminal vesicle gland coefficient decreased with increasing olaquindox concentration.There was no significant difference between the low OLA group and the blank control group.The OLA group was significantly lower than the blank control group(P<0.05),and the high OLA group was significantly lower than the blank control group(P<0.01).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.(3)The sperm density in the epididymis decreased with increasing olaquindox concentration.The high OLA group and the middle OLA group were significantly lower than the blank control group(P<0.05).There was no significant difference between the low OLA group and the blank control group.There was no significant difference between high OLA group and APS+VE group,VE group and APS group.The sperm viability rate showed a decreasing trend with increasing olaquindox concentration.There was no significant difference between the low OLA group and the blank control group.The middle OLA group was significantly lower than the blank control group(P<0.05).The high OLA group was significantly lower than the blank control group(P<0.01).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.The sperm motility showed a decreasing trend with increasing olaquindox concentration.The high OLA group was significantly lower than the blank control group(P<0.01),and the middle OLA group and low OLA group were significantly lower than the blank control group(P<0.05).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.Sperm deformity rate increased with the increase of olaquindox concentration.There was no significant difference between the low OLA group and the blank control group.The OLA group was significantly higher than the blank control group(P<0.05),and the high OLA group was significantly higher than the blank control group(P<0.01).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.The results showed that olaquindox poisoning resulted in slow growth and development of male mice,reduced testis coefficient and seminal vesicle glandular coefficient,decreased sperm density,decreased activity rate,and increased sperm deformity rate,and the worse the semen quality as the dose increased;VE,APS has no obvious protective effect on the slow development of testosterone,decrease of testis coefficient and seminal vesicle glandular coefficient,and decrease of semen quality.2.The effects of olaquindox,VE and APS interventions on the morphology of testis and epididymis in miceTests were performed on HE staining of mouse testis and epididymal tissue.(1)Testicular HE staining showed that: in the control group,the testicular tissue and seminiferous seminiferous tubules were arranged neatly;the spermatogenic cells at all levels were well-defined,closely arranged,the morphology of the interstitial cells was normal,and the basement membrane was complete and continuous.A lot of sperm.The structure and morphology of the seminiferous tubules in the testis of the low OLA group were normal,and there was no obvious morphological change compared with the blank control group.The morphological structure of the seminiferous tubules in the testis tissue of the Chinese OLA group showed obvious pathological changes,the number of spermatogenic cells decreased,the arrangement of spermatogenic cells at all levels was disordered,the number of spermatogenic cells decreased,the intercellular space increased,and the cells appeared swollen and vacuolated.Apoptosis,there is a small amount of cell necrosis material and a small amount of mature sperm in the lumen,basement membrane thinning,or even missing.The structure of the seminiferous tubules in the high OLA group was severely damaged,and the degeneration and necrosis of the spermatogenic cells at all levels increased significantly.The degree of apoptosis and vacuolization were extremely significant.There were a large number of necrotic substances and a few mature spermatozoa in the lumen,and the basement membrane became thin.,lack of serious.There was no significant difference between high OLA group and APS+VE group,VE group and APS group.(2)HE staining of epididymal tissue: The epididymis showed normal histological structure in the control group and the negative control group.The cells were closely arranged and regularly arranged.The cytoplasm of the epididymal head was full,showing a high columnar shape,and the cavity surface was more fine.Long stereocilia,a large number of mature spermatozoa in the epididymal gavage,and no obvious pathological changes in the epididymal epithelium of the low OLA group compared with the control group.The epithelial epithelium of the middle OLA group showed obvious pathological changes.The swelling and vacuolization of the cells were evident.Spermatozoa in the lumen decreased significantly,and the number of degraded cells increased.In the high OLA group,the histological lesions were aggravated,a large number of cells were swollen and vacuolated,cells were necrotic and detached,chromatin was concentrated,there was little or no spermatozoa in the lumen,the degree of vacuolation was significantly increased,and the static cilia was significantly shortened.There was no significant difference between the high OLA group and the APS group.APS+VE group,VE group were less injured than the high OLA group,and there was a small amount of sperm in the lumen.The results showed that the medium and high doses of olaquindox can cause pathological damage to the testis and epididymis structure of mice,destroy spermatogenic cells at all levels,and affect the normal function of the testis and epididymis.The high OLA group has extremely significant effects.Testicular tissue,epididymis tissue and cell damage are extremely serious.VE and APS had no obvious intervention on testicular structure,and VE had a slight protective effect on epididymal structure.3.The effects of olaquindox,VE and APS interventions on antioxidation of testis.The test examined the effects of olaquindox,VE,and APS interventions on the anti-oxidation function of testicular tissue by measuring SOD,GSH-Px activity and MDA,LPO content and SOD,GSH-Px expression levels in the testis.Test results show:(1)The activity of SOD increased with increasing concentration of olaquindox.The high OLA and middle OLA groups were significantly higher than the blank control group(P<0.01).There was no significant difference between the blank control group and the negative control group.The APS+VE group and VE group were significantly lower than the high dose(P<0.01),and the APS group was significantly higher than the high OLA group(P<0.01).The activity of GSH-Px decreased with the increase of olaquindox,and it was significantly lower in the middle OLA group than in the blank control group(P<0.05).The low OLA group and high dose were significantly lower than the blank control group and the negative control group(P<0.01).There was no significant difference between the high OLA group and the APS+VE group and the APS group.The VE group was significantly higher than the high OLA group(P<0.05).(2)The content of LPO increased with the increase of olaquindox concentration,and there was no significant difference between the low OLA group and the blank control group.The OLA group and high dose were significantly higher than the blank control group(P<0.05).There was no significant difference between the high OLA group and the VE group,the APS group,and the APS+VE group.The content of MDA increased with the increase of olaquindox,which was significantly higher in the low OLA group,the middle OLA group and the high OLA group than in the blank control group and the negative control group(P<0.05).There was no significant difference between low OLA group,middle OLA group and high OLA group.There was no significant difference between high OLA group and APS+VE group and APS group.VE group was significantly higher than high OLA group(P<0.05).(3)The expression of SOD decreased with the increase of olaquindox,but there was no significant difference between the middle OLA group,the low OLA group and X the blank control group.The high OLA group was significantly lower than the blank control group(P<0.05).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.The expression of GSH-Px increased with the increase of olaquindox concentration.Low OLA group,medium OLA group and high OLA group were significantly higher than the blank control group(P<0.01).The high OLA group was significantly higher than the APS+VE group,the VE group,and the APS group.The results showed that high-dose olaquindox caused severe oxidative stress in the testis,increased activity of antioxidant enzymes,increased production of oxidized products,decreased sperm quality,apoptosis of spermatogenic cells,and caused reproductive dysfunction.VE,APS had no obvious anti-oxidation protective effect on the reproductive system of mice.4.Effect of olaquindox,VE and APS intervention on gene expression of testosterone synthesis related enzymesThe test determined the expression levels of five testosterone synthase-related genes in testis 17-HSD,P450 scc,Star,3?-HSD,and P450c17 by real-time quantitative PCR.The effects of olaquindox,VE,and APS interventions on testosterone synthesis were analyzed.Test results show:(1)The expression of 17β-HSD showed a decreasing trend with increasing olaquindox concentration.The low OLA group and the middle OLA group were significantly lower than the blank control group and the negative control group(P<0.05).The high OLA group was significantly lower than the blank control group(P<0.01).There was no significant difference between high OLA group and APS+VE group,VE group and APS group.(2)The expression of P450 scc showed a decreasing trend with increasing olaquindox concentration.The OLA group and the high OLA group were significantly lower than the blank control group(P<0.05).There was no significant difference between the low OLA group and the blank control group.There was no significant difference between the high OLA group and the APS+VE group,the VE group and the APS group.difference.(3)The expression of Star showed a decreasing trend with increasing olaquindox concentration.The high OLA group was significantly lower than the blank control group(P<0.05).There was no significant difference between the middle OLA group,the low OLA group and the blank control group.There was no significant difference between the high OLA group and the APS+VE group,the VE group and the APS group.difference.(4)The expression of 3β-HSD increased first and then decreased with the increase of olaquindox,and was significantly lower in the low OLA group than in the control group(P<0.05).There was no significant difference between the OLA group,the high OLA group and the blank control group.The VE group,APS group,and APS+VE group were significantly lower than the high OLA group(P<0.01).(5)The expression of P450c17 decreased with the increase of olaquindox concentration.The high OLA group was significantly lower than the blank control group(P<0.05).There was no significant difference between the low OLA group,the middle OLA group and the blank control group,high OLA group.There was no significant difference between the VE group,the APS group,and the APS+VE group.The results showed that high-dose olaquindox inhibits the expression of testosterone synthesis-related enzyme genes in the testis of mice.The synthesis of testosterone in the body decreases,resulting in severe atrophy of the testis and seminal vesicles,severe decline in semen quality,and degeneration and necrosis of spermatogenic cells and epididymal epithelial cells.The structure and function of mice’s reproductive system are seriously damaged.VE and APS had no obvious protective effect on mouse testosterone synthesis.