Regulational Effects Of Il-10 And Morphine On Intracelular Thioredoxin Expression In Immune Cells

Thioredoxin (Trx) is a kind of small molecule protein which is expressed in various organisms. Trx system is comprised of the Trx, Trx reductase and NADPH. Although Trx has many biologic functions, its fundamental functions are regulating biologic redox equilibrium and preventing against oxidant injuries through its active site Cys-Gly-Pro-Cys. Thioredoxin binding protein-2 (TBP-2) is one kind of negative regulator of Trx. It can inhibit the redox activity of Trx and down regulate the expression of Trx.Following development in biology, the effects of redox regulation in cellular signal transduction have been one of the focuses of research. A lots of reports showed that redox regulation has important roles in immune response. The Trx system is just one of anti-oxidant systems, which include Cys, GSH system and SOD in organisms, moreover, many reports suggested that Trx indirectly participates in the regulations of immune response. However, the effects of Trx on immune regulation have not been known very well. One of the main reasons is that the immune system is complicated (including the polymorphism of various leukocytes).Since Trx has several effects in immune responses, which includes the effect on suppressing inflammation previously reported, this thesis mainly studied the relationship between Trx and IL-10. Secondly, the role of morphine stimulation on the Trx expression in mice spleens was investigated.In this study, the expression of Trx was detected by RT-PCR and Western blot in Jurkat cells (a kind of CD4+T cells) or U937 cells (a kind of monocytes) after IL-10 stimulating, and the expression of Trx in mice spleen was detected by Western blot after morphine administration.The quantitive PCR results indicated that the Trx mRNA were increased and the TBP-2 mRNA were decreased in Jurkat cells after IL-10 treatment for 6 h,12 h and 16 h. And protein measure showed that incubation with IL-10 for 16 h made the Trx increased in Jurkat cells while this kind of incease was partly blocked by PI3K inhibitor.Besides, other results suggested Trx expression is decreased by stimulated with IL-10 (20 ng/ml) in U937 cells at some time points (1 h,4 h,16 h); and pretreatment of PI3K inhibitor would block the suppressive effect of IL-10 on Trx expression in U937 cells. Finally, the experiments on morphine regulating the Trx expression in the mice spleens showed that the Trx expression was inhibitted in the male mice's spleens of Kunming, while female mates had reverse results in the urgent morphine administration. The Trx expression of the male C57BL/6 mice's spleens was also inhibited in the chronic morphine administration.