The Expression And Significance Of The Proline-rich Tyrosine Kinase In Systemic Lupus Erythematosus Lymphocytes

ObjectiveThrough detecting the expression levels of monocyte proline-rich tyrosine kinase (of Pyk2) in systemic lupus erythematosus (SLE), to study the role of Pyk2in the process of lymphocyte proliferation and the influence to protein signaling molecules Blys that promoteing B cell proliferation in lymphocytes. And to discuss the significance of Pyk2in the pathogenesis of systemic lupus erythematosus, obove these, to provide some experimental basis for the study of the pathogenesis of systemic lupus erythematosus, and a theoretical basis for the clinical use of drugs.Method1To take20ml fasting venous blood in patients with SLE activity group, the inactive group and the healthy group respectively early in the morning, mononuclear cells were isolated by lymphocyte separation medium and by density gradient centrifugation.2To detect the mRNA and cells levels of Pyk2in peripheral blood mononuclear cells (PBMCs) between SLE patients and healthy group by immunohistochemical method, real-time quantitative PCR.3The peripheral blood mononuclear cells (PBMCs) in SLE patients and healthy group were added to promoting lymphocyte proliferation agent (phorbol acetate, PMA) and Pyk2-specific inhibitor (TyrA9),was then to detect the influence of PMA and TyrA9on lymphocyte proliferation by CCK-8(Cell the CountingKit-8).4The peripheral blood mononuclear cells (PBMCs) in SLE patients and healthy group were added to promoting lymphocyte proliferation agent (phorbol acetate, PMA) and Pyk2-specific inhibitor (TyrA9),was then to detect the expression levels of small and medium-sized Gprotein (of Racl) and B cell stimulatory factor (Blys) in lymphocytic by semi-quantitative PCR.Result1Pyk2in the PBMCs of healthy group has a small amount of expression, the expression levels in PBMCs of patients with SLE activity group is higher than the group of inactive and healthy, the expression levels in PBMCs of patients with SLE inactive group is higher than the healthy group. Pyk2expression and activation of signaling proteins in PBMCs in SLE patients was significantly higher than the healthy group.2The mRNA expression of Pyk2:compared with the control group, Pyk2expression level in PBMCs of SLE was significantly increased. the difference between SLE activity group and inactive group was statistically significant (P<0.05),the difference between SLE activity group and the healthy group was statistically significant (P<0.05), between SLE inactive group and healthy group,the difference was statistically significant (P<0.05).3PMA and TyrA9on lymphocyte proliferation:PMA can promote the proliferation of PBMCs in SLE and healthy patients. After using TyrA9, the proliferation of PBMCs with SLE was inhibited, but the healthy group did not change significantly.4Blys and Racl expression levels in lymphocytes of SLE patients was significantly higher than healthy group:after using PMA, Blys and Racl expression levels were increased in SLE and healthy group, while using Pyk2-specific inhibitor to block of the activation of Pyk2,PMA could not induce the activation of the Blys and Racl in lymphocytes with SLE. For the healthy group, after using TyrA9to block of the activation Pyk2, PMA can stimulate proliferation of lymphocyte. So TyrA9has specific inhibition to lymphocytes in patients with SLE.Conclusion1Pyk2may be involved in the occurrence and development of SLE, and with SLE disease activity correlation.2Pyk2may be useful for the expression of PBMCs and the impact of the development of SLE, and abnormal expression of Pyk2on SLE has specificity.3Racl may be induced by Pyk2in patients with SLE lymphocyte proliferation and activation of signaling pathways important protein, Pyk2through the induction of Racl expression by T lymphocytes activation and abnormal value.4Pyk2was induced by PMA SLE in patients with PBMCs proliferation and promotes the expression of Blys and the activation of a key signaling proteins, the aberrant activation of B lymphocytes play a key role in the process of. Pyk2may be useful for Blys and B lymphocytes elicited aberrant expression, which further led to increased production of autoantibodies, eventually leading to the pathogenesis of SLE.5Pyk2may be useful for Racl and up-regulation of Blys and its receptor, thereby causing the abnormal expression of Blys, further promote B lymphocytes to produce autoantibodies, leading to the onset of SLE.