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Heroin is known to enhance catabolism and inhibit anabolism of purine nucleotides, leading to purine nucleotide deficiencies in rat brains. Here, we determined the effect of exogenous purine nucleotide administration on purine nucleotide metabolism in the brains of heroin-dependent rats. Heroin was administrated in increasing doses for 9 consecutive days to induce addiction, and the biochemical changes associated with heroin and purine nucleotide administration were compared among the treated groups. HPLC was performed to detect the absolute concentrations of purine nucleotides in the rat brain cortices. Furthermore, purine nucleotide administration alleviated the effect of heroin on purine nucleotide content, activity of essential purine nucleotide metabolic enzymes, and transcript levels of these genes. Our findings therefore represent a novel, putative approach to the treatment of heroin addiction. Opiates, such as morphine and heroin, are illegally used as recreational drugs worldwide, and can significantly impair the user's physical and mental health Chu et al. Prevalence of human immunodeficiency virus and its association with hepatitis B, C, and D virus infections among incarcerated male substance abusers in Taiwan. Risk behaviors, prevalence of HIV and hepatitis C virus infection and population size of current injection drug users in a China-Myanmar border city: results from a Respondent-Driven Sampling Survey in PLoS One, v. Evidence-based prevention interventions for people who use illicit drugs. Drug harms in the UK: a multicriteria decision analysis. Lancet, v. Therefore, the exploration of the effects of repeated exposure to these compounds is important for understanding the mechanisms of drug dependency and developing new treatment options. Role of G protein-coupled receptors GPCRs for purines and pyrimidines in mediating degeneration and regeneration under neuroinflammatory processes. Signal, v. DNA replication stress, genome instability and aging. Nucleic Acids Res. Mitochondrial DNA depletion syndromes: review and updates of genetic basis, manifestations, and therapeutic options. Neurotherapy, v. Impaired function of p53R2 in Rrm2b-null mice causes severe renal failure through attenuation of dNTP pools. Studies have shown that morphine and heroin can enhance the oxidation of xanthine and hypoxanthine in the lobus temporalis, lobus frontalis, and lobus parietalis of rats Liu et al. Morphine enhances purine nucleotide catabolism in vivo and in vitro. Acta Pharmacol. Heroin affects purine nucleotides catabolism in rats in vivo. Life Sci. Furthermore, it has been reported that morphine may increase UA concentrations in the corpus striatum, as well as in serum and extracellularly, in vitro Enrico et al. Effect of morphine on striatal dopamine metabolism and ascorbic and uric acid release in freely moving rats. Brain Res. Effect of Bacopa monniera on liver and kidney toxicity in chronic use of opioids. Phytomedicine, v. In patients where morphine was administered intracerebroventricularly for cancer-related pain, the UA levels in cerebrospinal fluid samples increased significantly Goudas et al. Acute decreases in cerebrospinal fluid glutathione levels after intracerebroventricular morphine for cancer pain. Differential effects of acute morphine administrations on polymorphonuclear cell metabolism in various mouse strains. Previously, we have shown that opioids may lead to nucleotide metabolism disorders. Rats also received heroin ip over a period of 9 days in doses of 3, 3. Significant increases in UA concentration in plasma were observed in the rats following both morphine and heroin administration. The enzymatic activities of ADA and XO were found to be enhanced significantly in the brain and other organs of rats administered heroin, and morphine was shown to increase ADA and XO gene expression in the rat brain cortex Liu et al. Our experiments using rat C6 glioma cells also revealed that the gene expression of HGPRT and AK, two key enzymes in the purine nucleotide salvage pathway, are down-regulated by morphine, indicating that morphine inhibits purine nucleotide anabolism Liu, Hong, Zhao, LIU, J. Zhonghua Yi Xue Za Zhi, v. To further clarify the effects of heroin on purine nucleotide metabolism in vivo , we measured the absolute content of purine nucleotides in rat brain cortices by HPLC, and found that the levels of AMP and GTP are significantly reduced in rats administered heroin, resulting in a purine nucleotide deficiency Li et al. Heroin affects purine nucleotides metabolism in rat brain. Therefore, we hypothesize that purine nucleotide deficiency in the brain may be one of the biochemical mechanisms of heroin dependence. Thus, the goals of this study were to confirm that purine nucleotide metabolism disorders are induced by heroin, and to clarify whether purine administration can counteract the biochemical changes caused by heroin. Each group included 10 animals, and heroin was administered twice a day at h intervals. Group I served as the control group and rats in this group received normal saline ip for 9 days. Heroin was administrated ip in increasing daily doses of 0. Effect of purine nucleotides on conditioned place preference and acute withdrawal in chronic heroin dependent rats. On the morning of the 10th day, animals were killed via decapitation following ether-induced narcosis. Their brains were rapidly removed and thoroughly washed to remove the excess blood in ice-cold saline. All other chemicals used in this study were of analytical grade. Nanjing, China. Dalian, China. Development of an ion-pair HPLC method for investigation of energy charge changes in cerebral ischemia of mice and hypoxia of Neuro-2a cell line. Cortex tissue 0. The tissue samples were sonicated in 0. The supernatant was filtered through 0. Linearity was tested using five known standard solutions of increasing concentrations. Calibration curves were calculated by linear regression analysis of the peak area vs. Protein content was estimated using Coomassie blue protein detection kits. The activity of ADA and XO in the cortex samples were estimated using Guisti colorimetric and colorimetric detection kits, respectively Li et al. The primers were synthesized by the Jinsite Corporation Nanjing, China. All samples were run in well optical plates in duplicate. The cycle threshold values Ct were calculated and exported to Microsoft Excel for analysis. Quantification was performed by experimental determination of Ct, defined as the cycle at which the fluorescence exceeds 10 times the standard deviation of the mean baseline emission for the early cycles Collantes-Fernandez et al. Quantitative detection of Neospora caninum in bovine aborted fetuses and experimentally infected mice by real-time PCR. Statistical analyses were performed using SPSS Significance was applied to values of 0. The mean XO enzyme activity of each of the four groups was ADA and XO are essential enzymes for purine catabolism. ADA irreversibly catalyzes the hydrolysis of adenosine to inosine. Inosine is then deribosylated by purine nucleoside phosphorylase, converting it to hypoxanthine Cristalli et al. Adenosine deaminase: functional implications and different classes of inhibitors. XO catalyzes the oxidation of hypoxanthine and xanthine to UA. These results indicate that heroin increases the ADA and XO levels in the cortex, and leads to increased purine nucleotide catabolism. Our data show significant increases in the ADA and XO mRNA levels in the cortices of rats administered with heroin alone, as compared with those in the control group that received only saline. Our results also show that these increases were counteracted by purine nucleotide administration. Therefore, we conclude that the effect of heroin on purine nucleotide metabolism is dependent on the level of gene expression. Purine nucleotides are synthesized either from simple precursors by de novo synthesis through energetic, multistep reactions, or assembled from free purine bases like guanine, hypoxanthine, and adenine, through nucleotide salvage pathways Camici et al. Pediatric neurological syndromes and inborn errors of purine metabolism. However, in some organs, including the brain, purine nucleotide synthesis relies exclusively on salvage pathways. Inborn errors of purine metabolism: clinical update and therapies. Inherit Metab. Metabolic network of nucleosides in the brain. Our results reveal that heroin inhibits the transcription of these three key enzymes involved in the purine nucleotide synthesis pathway in the rat cortex, however, the inhibitory effects of heroin on these enzymes could be negated with the administration of exogenous purine nucleotides. Liquid chromatographic analyses were performed as previously described. Figure 4 shows the retention time and area of the peak of the mixed reference solutions. Figure 5 shows the calibration curve obtained from cortex tissue for the purine nucleotides. Figure 6 shows the HPLC chromatogram of cortex tissue. The AMP and GTP content in the group administered heroin decreased significantly as compared with those in the saline-treated group, which indicates that heroin reduces the content of some purine nucleotides in brain. Purine nucleotides play an important role in DNA synthesis, energy metabolism regulation, protein synthesis and function, and enzymatic activity. Genetic and metabolomic analysis of AdeD and AdeI mutants of de novo purine biosynthesis: cellular models of de novo purine biosynthesis deficiency disorders. It has been reported that biochemical changes such as decreased GMP concentrations are associated with opioid dependence in humans Mannelli et al. Opioid use affects antioxidant activity and purine metabolism: preliminary results. It has also been shown that opioid use may have toxic effects on the central and peripheral nervous systems, leading to irreversible, pathological cellular changes. The opioid growth factor-opioid growth factor receptor axis regulates cell proliferation of human hepatocellular cancer. R, The opioid growth factor-opioid growth factor receptor axis: homeostatic regulator of cell proliferation and its implications for health and disease. Our results indicate that the effect of heroin on the metabolism of purine nucleotides may result in purine nucleotide deficiency in the brain, representing a putative biochemical mechanism of heroin addiction. Our results also show that external compensation of purine nucleotides alleviates the deficiency in purine nucleotides in rat cortices caused by heroin, although purine nucleotide administration alone does not alter purine nucleotide concentrations, essential enzyme activity, or the mRNA levels of these key enzymes. Thus, purine nucleotide administration may represent a novel, effective approach to treating heroin addiction. Our data show that heroin promotes catabolism and inhibits anabolism of purine nucleotides in the rat cortex, resulting in a purine nucleotide deficit. However, the administration of exogenous purine nucleotides can reverse these changes in nucleotide metabolism. Although our results are preliminary, they indicate that the relationship between purine nucleotide metabolism and opioid addiction deserves further study. Open menu Brazil. Brazilian Journal of Pharmaceutical Sciences. Open menu. Text EN Text English. CHEN, Y. CHU, F. LI, K. LI, L. LIU, C. LIU, J. NUTT, D. YANG, Y. Publication Dates Publication in this collection Dec History Received 26 Nov Accepted 09 Sept This is an open-access article distributed under the terms of the Creative Commons Attribution License. Department of Pharmacology. College of Basic Medical Sciences. E-mail: lik jlu. Figures 6 Tables 2. Lineu Prestes, n. Stay informed of issues for this journal through your RSS reader. PDF English. Google Google Scholar. Effects of purine nucleotide administration on purine nucleotide metabolism in brains of heroin-dependent rats.

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