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Morphine Induces Splenocyte Apoptosis and Enhanced mRNA Expression of Cathepsin-B

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Abstract

Morphine has been demonstrated to modulate immune function. We studied whether morphine modulates apoptosis of splenocytes. Splenocytes were isolated from control and morphine treated rats. Splenocytes isolated from morphine treated rats showed increased percentage (P < 0.001) of apoptosis when compared to splenocytes isolated from untreated rats (control, 4.7 ± 1.0% apoptotic splenocytes/field vs. morphine, 47.8 ± 3.4% apoptotic splenocytes/field). These results were further confirmed by gel electrophoresis as well as by end-labeling DNA of splenocytes isolated from control and morphine treated rats. Splenocytes from morphine treated rats showed a classical ladder pattern with integer multiples of 180 base pairs. Splenocytes from morphine treated rats also showed increased mRNA expression of cathepsin-B, a gene associated with active cell death. These results suggest that morphine may also be modulating immune function by enhancing apoptosis of splenocytes.

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REFERENCES

  1. LOURIA, D. B., T. HENSLE, and J. ROSE. 1957. The major medical complications of heroin addiction. Ann. Intern. Med. 67:1–22.

    Google Scholar 

  2. BRIGGS, J. H., C. G. MCKERRON, R. I. SOUHAMI, D. J. E. TAYLOR, and H. ANDREWS. 1967. Severe systemic infections complicating mainline heroin addiction. Lancet 2:1227–1231.

    Google Scholar 

  3. CHERUBIN, C. E. 1971. The medical sequelae of heroin addiction. Arch. Intern. Med. 128:309–313.

    Google Scholar 

  4. TUBARO, E., G. BORELLI, C. CROSE, G. CAVALLO, and C. SANTIANGELI. 1983. Effect of morphine on resistance to infection. J. Infect. Dis. 148:655–666.

    Google Scholar 

  5. MC DONAUGH, R. J., J. J. MADDEN, A. FALEK, D. A. SHAFER, M. PLINE, D. GORDON, P. BOKOS, J. C. KUEHNLE, and J. MENDELSON. 1980. Alteration of T and null lymphocyte frequencies in the peripheral blood of human opiate addicts. J. Immunol. 125:2539–2543.

    Google Scholar 

  6. BROWN, S. W., B. STIMMEL, R. N. TAUB, S. KOCHWA, and R. E. ROSENFIELD. 1974. Immunologic dysfunction in opiate addicts. Arch. Intern. Med. 134:1001.

    Google Scholar 

  7. WYBRAN, J., T. APPELBOOM, J. P. FAMAEY, and A. GOVAERTS. 1979. Suggestive evidence for receptors for morphine and methionine-enkephalin on normal human blood T lymphocytes. J. Immunol. 123:1068–1070.

    Google Scholar 

  8. ARORA, P. K., E. FRIDE, J. PETTITO, K. WAGGIE, and P. SKOLNICK. 1990. Morphine-induced immune alterations in vivo. Cell Immunol. 126:343–353.

    Google Scholar 

  9. BRYANT, H. U., E. W. BERNTON, and J. W. HOLADAY. 1987. Immunosuppressive effects of chronic morphine treatment in mice. Life Sci. 41:1731–1738.

    Google Scholar 

  10. SEI, Y., K. YOSHIMOTO, T. MCINTYRE, P. SKOLNICK, and P. K. ARORA. 1991, Morphine-induced thymic hypoplasia is glucocorticoid-dependent. J. Immunol. 146:194–196.

    Google Scholar 

  11. FUCHS, B. A., and S. B. PRUETT. 1993. Morphine induces apoptosis in murine thymocytes in vivo but not in vitro: Involvement of both opiate and glucocorticoid receptors. J. Pharmacol. Exp. Ther. 266:417–423.

    Google Scholar 

  12. THOMPSON, C. B. 1995. Apoptosis in the pathogenesis and treatment of disease. Science 267:1456–1462.

    Google Scholar 

  13. WYLLIE, A. H., J. F. R. KERR, and A. R. CURRIE. 1980. Cell death: The significance of apoptosis. Int. Rev. Cytol. 68:251–305.

    Google Scholar 

  14. SAVILL, J., V. FADOK, P. HENSON, and C. HASLETT. 1993. Phagocytic recognition of cells undergoing apoptosis. Immunol. Today 14:131–136.

    Google Scholar 

  15. OBERHAMMER, F., P. R. WILSON, and M. SIKORSKA. 1993. Apoptotic death in epithelial cells: Cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J. 12:3679–3684.

    Google Scholar 

  16. FLIPSKI, J., J. LEBLANC, T. YOUDALE, M. SIKORSKA, and P. R. WALKER. 1990. Periodicity of DNA folding in higher order chromatin structures. EMBO J. 9:1319–1327.

    Google Scholar 

  17. BROWN, D. G., X. M. SUN, and G. M. COHEN. 1993. Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation. J. Biol. Chem. 268:3037–3039.

    Google Scholar 

  18. WYLLIE, A. H. 1980. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284:555–556.

    Google Scholar 

  19. MARTIN, S. J., and D. R. GREEN. 1995. Protease activation during apoptosis: Death by a thousand cuts? Cell 82:349–352.

    Google Scholar 

  20. NICHOLSON, D. W., A. ALI, N. A. THORNBERRY, J. P. VALLIANCOURT, C. K. DING, M. GALLANT, Y. GAREAU, P. R. GRIFFIN, M. LABELLE, Y. A. LAZEBNIK, N. A. MUNDAY, S. M. RAJU, S. E. SMULSON, T. T. YAMIN, V. L. YU, and D. K. MILLER. 1995. Identification and inhibition of the ICE/ced-3 protease necessary for mammalian apoptosis. Nature 376:37–43.

    Google Scholar 

  21. MICHEL, D., J. G. CHABOT, E. MOYSE, E. MOYSE, M. DANIK, and R. QUIRION. 1992. Possible functions of a new genetic marker in central nervous system: the sulfated glycoprotein-2 (SGP-2), Synapse 11:105–111.

    Google Scholar 

  22. GUENETTE, R. S., M. MOOIBROEK, K. WONG, P. WONG, and M. TENNISWOOD. 1994. Cathepsin-B. A cysteine protease implicated in metastatic progression, is also expressed during regression of the rat prostate and mammary glands. Eur. J. Biochem. 226:311–321.

    Google Scholar 

  23. LIEBERTHAL, W., V. TRIACA, and J. LEVINE. 1996. Mechanism of death induced by cysplatin in proximal tubular epithelial cells: apoptosis vs. necrosis. Am. J. Physiol. 270:F700–F708.

    Google Scholar 

  24. CHOMCZYNSKI, P., and N. SACCHI. 1987. Single-step method of RNA isolation by acid guanidine thiocynate phenol-chloroform extraction. Anal. Biochem. 162:156–159.

    Google Scholar 

  25. SHAVIT, Y., A. DEPAULIS, F. C. MARTIN, G. W. TERMAN, R. P. GALE, and J. C. LIEBESKIND. 1986. Involvement of brain opiate receptors in the immune suppressive effect of morphine. Proc. Natl. Acad. Sci. U.S.A. 83:7114–7117.

    Google Scholar 

  26. WEBER, R. J., and A. PERT. 1989. The periaqueductal gray matter mediates opiate-induced immune suppression. Science 245:188–190.

    Google Scholar 

  27. HO, W. K. K., and A. LEUNG. 1979. The effect of morphine addiction on concanavalin A-mediated blastogenesis. Pharmacol. Res. Commun. 11:413–419.

    Google Scholar 

  28. BRYANT, H. U., E. W. BERNTON, and J. W. HOLADAY. 1987. Immunosuppressive effects of chronic morphine treatment in mice. Life Sci. 41:1731–1738.

    Google Scholar 

  29. BRYANT, H. U., E. W. BERNTON, and J. W. HOLADAY. 1988. Morphine pellet-induced immunomodulation in mice: Temporal relationship. J. Pharmacol. Exp. Ther. 245:913–920.

    Google Scholar 

  30. BRYANT, H. U., E. W. BERNTON, J. R. KENNER, and J. W. HOLADAY. 1991. Role of adrenal cortical activation in the immunosuppressive effects of chronic morphine treatment. Endocrinology 128:3253–3258.

    Google Scholar 

  31. BRYANT, H. U., and R. E. ROUDEBUSH. 1990. Suppressive effects of morphine pellet implants on in vivo parameters of immune function. J. Pharmacol. Exp. Ther. 255:410–415.

    Google Scholar 

  32. WEBER, R. J., B. IKEJIRI, K. C. RICE, A. PERT, and A. A. HAGAN. 1987. Opiate receptor-mediated regulation of the immune response in vivo. NIDA Res. Monogr. 76:341–348.

    Google Scholar 

  33. BAYER, B. M., M. R. GASTONGUAY, and M. C. HERNANDES. 1992. Distinction between the in vitro and in vivo inhibitory effects of morphine on lymphocyte proliferation based on agonistic sensitivity and naltrexone reversibility. Immunopharmacology 23:117–124.

    Google Scholar 

  34. SEI, Y., T. MC INTYRE, E. FRIDE, K. YOSKIMOTO, P. SKOLNICK, and P. K. ARORA. 1991. Inhibition of calcium mobilization is an early event in opiate-induced immunosuppression. FASEB J. 5:2194–2199.

    Google Scholar 

  35. LOPHER, A., L. G. ABOOD, W. HOSS, and F. J. LIONETTI. 1980. Steroselective muscarinic acetylcholine and opiate receptors in human phagocytic leucocytes. Biochem. Pharmacol. 29:1361–1365.

    Google Scholar 

  36. MEHRISHI, J. N., and I. H. MILLS. 1983. Opiate receptors on lymphocytes and platelets in man. Clin. Immunol. Immunopathol. 27:240–249.

    Google Scholar 

  37. MADDEN, J. J., R. M. DONAHOE, J. ZWEMER-COLLINS, D. A. SHAFER, and A. FALEK. 1987. Binding of naloxone to human T lymphocytes. Biochem. Pharmacol. 36:4103–4109.

    Google Scholar 

  38. OVAIDA, H., P. NITSAN, and O. ABRAMSKY. 1989. Characterization of opiate binding sites on membranes of rat lymphocytes. J. Neuroimmunol. 21:93–102.

    Google Scholar 

  39. OVAIDA, H., and O. ABRAMSKY. 1991. Opiate binding sites on rat lymphocytes. Prog. Neuroenodocrin. Immunol. 4:121–125.

    Google Scholar 

  40. RADULESCU, R. T., B. R. DE COSTA, A. E. JACOBSON, K. C. RICE, J. E. BLALOCK, and D. J. J. CARR. 1991. Biochemical and funtional characterization of a μ-opioid receptor binding site on cells of the immune system. Prog. Neuroendocrin. Immunol. 4:166–179.

    Google Scholar 

  41. TAUB, D. D., T. K. EISENSTEIN, E. B. GELLER, M. W. ADLER, and T. J. ROGERS. 1991. Immunomodulatory activity of kappa-, mu, and delta-selective opioid agonists. Proc. Natl. Acad. Sci. U.S.A. 88:360–364.

    Google Scholar 

  42. ROGERS, T. J., D. D. TAUB, T. K. EISENSTEIN, E. B. GELLER, and M. W. ADLER. 1991. Immunomodulatory activity of kappa-, mu-, and delta-selective opioid compounds. NIDA Res. Monogr. 105:82–88.

    Google Scholar 

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Singhal, P.C., Reddy, K., Franki, N. et al. Morphine Induces Splenocyte Apoptosis and Enhanced mRNA Expression of Cathepsin-B. Inflammation 21, 609–617 (1997). https://doi.org/10.1023/A:1027334122387

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