| Objectives:In recent years,the abuse of the new synthetic drugs represented by METHamphetamine(METH)has become increasingly serious,and the number of drug abusers has drastic increased.Because of its strong central excitability and addiction,long-term abusing could easily lead to organ damage,mental illness and crime.It can cause serious social public health problems,and as the trend of younger abuser is increasing,and METH damages the reproductive system,which has received more and more attention.In this study,adult male rats were injected intraperitoneally with METH to study the effect on the reproductive function,the expression changes of glandular hormone levels,androgen and estrogen receptors.In order to provide a theoretical basis for the effects of METH abuse on male reproductive gonadal hormone and testosterone receptors.METHods:To investigate the damage of METH on the reproductive system in male rats,a model of METH exposure was established by intraperitoneal injection of METH in adult male rats.Forty SD male rats were randomly divided into the acute control group,acute exposure group,chronic control group and chronic exposure group,with 10 rats in each group.In order to confirm the the reliability of the model,we observed the general situation of the METH exposure group,whether there was stereotyped behavior,scored and recorded the duration time.In order to study the damage of METH on rat testis and sperm,after the rats were anesthetized and the testis were harvested,the testis were weighed and the testicular index changes were calculated.The changes of spermatogenic cell structure in testis tissue were observed by hematoxylin-eosin(HE)staining,and the epididymis was obtained for semen quality testing.To analyze the related gland hormone levels in the serum of rats,we performed enzyme-linked immunosorbent assay(Elisa)to detect the levels of testosterone(T),estradiol 2(E2),follicle stimulating hormone(FSH),luteinizing hormone Expression levels of(LH),progesterone(PROG),prolactin(PRL)and inhibin B(INH B).To quantify and confirm the expression and distribution of androgen and estrogen receptors,the immunohistochemical staining(IHC),quantitative real-time polymerase chain reaction(QRT-PCR)and Western blot(WB)were performed to investigate the localization and expression changes of androgen receptor(AR),estrogen receptor alpha(ERa)and estrogen receptor beta(ER?)in the testis.Results:After the administration with METH,the general situation observation of the rats in the METH-exposure group was performed the increasing alertness,repeated nodding and exploring and other repeated stereotypes behavior,the stereotypic behavior scores of the acute and chronic exposure groups were significantly increased in comparison of the acute and chronic control groups(P<0.05),confirming that the model was successfully established.By measuring body weight and bilateral testicular weight,the body weight of rats in acute and chronic exposure group was significantly lower than that in the control group(P<0.05),and the testicular index of rats in chronic exposure group was lower than that in the chronic control group(P<0.05).The results of HE staining showed the testicular spermatogenic cells changed in the acute and chronic exposure groups,the spermatogenic cells were reduced and disordered.In the chronic exposed group,there is a marked emergence of germ cell shedding and no sperm formation in the lumen.Semen quality analysis showed that the concentration of epididymal semen and total viability in the chronic exposure group were significantly lower than those in the control group(P<0.05).These results suggest that METH severely affects the spermatogenic function of SD rats.To investigate the effect of METH exposure on gonadal axis hormones in SD rats,Elisa results showed that T values were significantly decreased in the acute and chronic exposed groups in comparison of the control group(P<0.05,P<0.01);compared with the chronic control group,the E2 and INH B were decreased significantly in the chronic exposure group(P<0.05),while the PRL was increased(P<0.01).In addition,in order to study the effect of METH on the expression of AR,ERa and ER? in SD rats,IHC results suggested that AR was distributed in Leydig cells,supporting cells,in the nucleus of spermatogonia,without the expression of sperm;ER is widely distributed in the spermatogenic cells in the testis,and the integrated optical density values indicate that AR,ERa,and ER? were significantly decreased in the acute and chronic exposure groups.Similarity,the results of QRT-PCR and WB showed that the mRNA and protein expression of AR,ERa and ER?in the testis were significantly decreased in the acute and chronic exposure group in comparison of the control group.Conclusion:We found that long-term METH exposure can impair the spermatogenic function via the intraperitoneal injection of METH in male rats;it may be caused by a decrease in the level of gonadotropin and the expression of the testosterone receptors AR,ERa,and ER? in male rats. |
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