Effect Of Heroin And Purine Nucleotide Compensation On Reproductive Endocrine System Of Male Rats

In recent years, opiate addiction and the abuse has become a serious medical and social issues. At present, as one of the popular major opioid drug, heroin, whether long-term or short-term was used, can cause damage and potential toxicity of cardiovascular, nervous system, kidney, reproductive endocrine system and immune system. but there is still a lack of ideal drug treatment up to now. The study on opiate drug addiction, physical dependence and mental dependence, and tolerance of the drug mechanism is the subject that scientific workers still to do constantly. Based on the reason that men is the major part of heroin addict and there is still gender differences in opioid abuse, the paper explored the effect of heroin on male reproductive endocrine function and the the changes when purine nucleotides were compensated. This paper is also based on the our laboratories relevant past work that morphine and heroin can enhance purine nucleotide catabolism of organizations and weak their salvage synthesis.Heroin dependence, heroin withdrawal and purine nucleotides (AMP and GMP ) compensation male rat models were established. After success modeling, rats were randomly divided into saline control group(C), nucleotide control group(N), heroin treatment group(H), heroin + nucleotide group (NH), withdrawal 3 d group(W3), withdrawal 9 d group(W9), nucleotide 3d group(N3) and nucleotide 9d group (N9).1. Effects of heroin and purine nucleotide compensation on reproductive hormones in rat plasmaTestosterone(T), follicle stimulating hormone(FSH)and luteotrophic hormone (LH)/ interstitial cell-stimulating hormone(ICSH) is very important indicator that can response the hypothalamus-pituitary-testicular axis function. The changes of these hormones in the plasma ware detected by radioimmunoassay when heroin was acted and purine nucleotides were compensated.Plasma testosterone levels of H group have a downward trend, rebounded after the withdrawal, but there is no significant difference compared with the C group. There is a also lower testosterone levels in NH group. After the administration of heroin,plasma FSH levels were significantly decreased and rebound after the withdrawal. And plasma FSH levels also decreased significantly in NH,N3,N9 groups. Compared with H group, plasma FSH levels in W9 group increased significantly (p< 0.01). Plasma LH have a downward trend after heroin administration and rebounded after the withdrawal, LH levels are lower in NH group, but they had no significant statistical difference (p> 0.05),compared with C group.The results showed that short-term heroin can induce the HPG axis function in disbalance. And the impact of FSH is more pronounced, impact of testosterone and LH is not yet significant. And the role of heroin is durative, will not immediately disappear after the heroin withdrawal. At the same time, purine nucleotide compensation can remaine the plasma reproductive hormones FSH,LH and testosterone levels relatively stable. So the result suggests that purine nucleotide compensation may inhibit adverse effects of heroin on male HPG axis of rats.2. Effect of heroin and purine nucleotide compensation on FSH, LH gene expression of rat pituitaryIn order to further explore the effect of heroin and purine nucleotide compensation on reproductive endocrine system, rat pituitary FSH and LH in the gene expression level was detected using RT-PCR method.After the administration of heroin, FSH and LH mRNA relative expression have a downward trend and rebounded after withdrawal. FSH and LH mRNA expression of the relative expression in NH, H3 and H9 are significantly higher than saline control group and heroin group (p <0.01).This study found heroin has no significant effect on FSH and LH expression. Purine nucleotide compensation can enhance LH, FSH expression of rat pituitary , both in the period of heroin addiction or withdrawal.3. Effect of heroin and purine nucleotide compensation on adrenocorticotrophic hormone (ACTH)in rat plasmaACTH is one of the product of stress and nervous system regulating factor. It play an important role in the maintenance of the growth and secretion functions of adrenal. Its content is important evaluation index of the HPA axis function of the body.The results showed that plasma ACTH of H group reduced significantly, and plasma ACTH of W9 group decreased more significantly. Compared with the H group, the plasma ACTH of N3 group increased significantly (p <0.05). The results suggest that heroin affects plasma ACTH of male rats, and role of heroin has a certain continuity. Purine nucleotide compensation can maintain plasma ACTH in the relatively stable.4. Effect of heroin and purine nucleotide compensation on rat epididymis neutralα-glucosidaseEpididymis is mature and storage sites of male animals sperm. Neutralα-glucosidase is specific enzymes and marker of the epididymis, can be used to reflect the function of the epididymis. The results showed that the heroin has dropped the neutralα-glucosidase content of rat epididymis. It is still low after heroin withdrawal. Purine nucleotide compensation can maintain neutralα-glucosidase in the relatively stable. Compared with H group, neutralα-glucoside activity of NH group increased (p <0.05).The results showed that heroin decreases neutralα-glucoside activity in the epididymis, and will continue for some time after withdrawal, can not be recovered immediately. Purine nucleotide compensation likely to improve the functional status of the epididymis, but the mechanism is more complex, and need to study further.5. Effect of heroin and purine nucleotide compensation on key enzyme of purine nucleotide metabolism in rat testis and epididymis, pituitaryPurine nucleotide catabolism key enzyme adenosine deaminase(ADA)and purine nucleotide salvage synthesis key enzyme hypoxanthine- guaninephosphoribosyl transferase (HGPRT) for research object, to study the impact of heroin and purine nucleotides (AMP and GMP)compensation on purine nucleotide metabolism key enzyme of the rat testis,epididymis and pituitary.A.Testis and epididymis:Compared with C group, ADA activity increased (p<0.05) of H, W3 groups in the testis and epididymis. Compared with H group, the ADA activity of NH group in testis and epididymis declined, but the extent is different.Compared with C group, the ADA mRNA relative expression of H group in rat testis and epididymis increased significantly (p<0.05).Compared with H group, the ADA mRNA relative expression of NH,W9,N3 and N9 Group in rat testis and epididymis declined significantly.The HGPRT mRNA relative expression of H group in rat testis increased significantly, compared with C group (p<0.05). And the HGPRT mRNA relative expression of N,NH,W3,W9,N3 and N9 groups were lower than that of H group (p< 0.05).The HGPRT mRNA relative expression of H,N,NH,W3 and N9 groups in rat epididymis increased significantly(p<0.05), compared with C group. And the HGPRT mRNA relative expression of NH and N9 groups in rat epididymis were higher than that of H group (p<0.05). B. Pituitary:The ADA mRNA relative expression of NH group in pituitary is lower significantly (p<0.05), than that in H group,. The HGPRT mRNA relative expression of N group in rat pituitary increased significantly (p<0.05), compared with C group.And the HGPRT mRNA relative expression of W3 group in pituitary increased significantly (p<0.05), compared with H group. Heroin can enhance key enzyme ADA activity of purine nucleotide catabolism in testis and epididymis, and heroin has a certain continuity. And at the same time, purine nucleotide compensation"oppose"the role of the heroin.The impact of heroin and purine nucleotide compensation on synthetic remedies key enzyme HGPRT in the testis and epididymis seems different,but the reason is not clear.Purine nucleotide (AMP and GMP), as composition of RNA / DNA, is very important within the cell. At the same time, purine nucleotide are neurotransmitters, involved in almost all activities of the lives. Therefore, the discovery of the relationship between opiates and purine nucleotide metabolism may be a new interpretation for the body and spirit damage of the abuse of opiate addicts, and also clarify the male reproductive endocrine effects of heroin from a new perspective.6. Effect of heroin and purine nucleotide compensation on metabolism of glutamate of rat testis and epididymisBy detecting the content of glutamate(Glu), and the content of glutamate metabolism key enzyme glutamate dehydrogenase(GDH)and glutamine synthetase(GS), to study the impact of heroin and purine nucleotides(AMP and GMP) compensation on glutamic acid metabolism enzyme content of the rat testis and epididymis.A. TestisCompared with C group, Glu of H group in testis were significantly increased, and Glu of W3, W9 and NH groups in testis decreased (p<0.05). Compared with H group, Glu of NH, W3, W9, N3, N9 groups were significantly decreased (p <0.05, p< 0.01). Heroin made a downward trend Of GDH and GS activity in testis, and GDH and GS activity rebounded after heroin withdrawal. GDH activity of W9 group was significantly higher than that of H group, GS activity was significantly higher than that in C and H groups (p <0.05).B.epididymis:Compared with C group, Glu content of H, W3, W9 groups in epididymis increased, and Glu of NH group decreased ,but there is no significant difference (p>0.05). Compared with the C group, GDH activity of H group in epididymis decreased significantly (p<0.05). GDH activity of NH, W3, W9, N3 and N9 groups increased significantly (p<0.05, p<0.01), compared with H group. GS activity of H, W3, NH and N9 groups decreased significantly (p<0.05, p<0.01), compared with C group. Compared with H group, GS activity of NH group in epididymis decreased significantly (p<0.05).The results showed there is trend that heroin inhibit GDH and GS activity in rat testis and epididymis, and the overall trend is an increasing of Glu. At the same time, purine nucleotide compensation or withdrawal can reversed the effect on Glu metabolismof heroin in rat testis and epididymis.7. Effect of heroin and purine nucleotide compensation on gonadotropin cell morphology of pituitaryGonadotropin cell within the pituitary is an important endocrine cell, it secrete FSH and LH. The results showed that gonadotropin cell of C group: lager cell, larger and round or oval-shaped cell nuclear, more euchromatin in cell nuclear;round secretory granules, medium-sized electron density and small size secretory granules, rich mitochondria, rough endoplasmic reticulum and Golgi complex. Gonadotropin cell of H group: cell nuclear is irregular in shape, the heterochromatin in nuclear increased, rough endoplasmic reticulum expansion slightly, cytoplasm cavity structure markedly increased,the number of secretory granules reduced, the electron density is low. Gonadotropin cell of NH group: cell structure closer to C group, a round of nuclear, nuclear matrix slightly empty; the number of endocrine particles increased than in H group, but less than in C group. The result shows that heroin damage gonadotropin cells ultrastructure of pituitary, purine nucleotide compensation can ease or inhibit this damage.