Study On The Function And Mechanism Of P2Y₆ In Systemic Lupus Erythematosus

Objective:Systemic lupus erythematosus(systemic lupus erythematosus,SLE)is a typical autoimmune disease with chronic multi-system and multiple organ damage,which is characterized by the formation of a large number of autoantibodies and complements,which is seriously harmful to human health.In recent years,it has been found that its occurrence is closely related to aseptic inflammation induced by tissue injury,in which the increased infiltration and abnormal activation of tissue inflammatory cells play a key role in the occurrence and development of the disease.Studies have shown that purine receptor P2Y6 is highly expressed in inflammatory cells and plays a pro-inflammatory role in regulating the infiltration of inflammatory cells and the release of inflammatory factors.However,the role of P2Y6 receptor in SLE has not been reported,so this paper mainly studies the regulatory role of purine receptor P2Y6 in the occurrence and development of SLE,and provides new targets and new ideas for clinical immunotherapy of SLE.Methods:1.Real-time fluorescence quantitative PCR was used to analyze the difference of P2Y6 expression in peripheral blood mononuclear cells between healthy people and SLE patients.2.The lupus model of mice was established by intraperitoneal injection of Pristane.Anti-double-stranded DNA antibody(anti-dsDNA antibody)and anti-small nuclear ribonucleoprotein antibody(anti-snRNP antibody)in serum of lupus mice were detected by enzyme-linked immunosorbent assay(ELISA),and the effects of serum total IgG and IgM,on the production of autoimmune antibodies in lupus mice were analyzed.3.immunofluorescence staining were used to detect the effects of P2Y6 KO on glomerular IgG and complement C3 deposition in lupus mice.4.Flow cytometry was used to detect the effect of P2Y6 KO after 2 weeks of Pristane injection on the exudation ratio and surface activation/maturation markers of Ly6Chi monocytes in abdominal cavity of lupus mice.5.Flow cytometry was used to detect the effects of P2Y6 KO on the proportion of T and B lymphocytes and T cell activation in spleen of lupus mice after 8 months of Pristane injection.Mouse bone marrow cells were induced to mature in the medium containing L929 supernatant.The effect of P2Y6 receptor on the differentiation and maturation of mouse bone marrow cells was detected by flow cytometry.Results:1.Compared with healthy subjects,the expression of P2Y6 in(PBMC)of peripheral blood mononuclear cells in patients with SLE was increased;2.The mouse model of systemic lupus erythematosus was established successfully.it was found that P2Y6 KO could significantly reduce the contents of serum anti-double-stranded DNA antibody(anti-dsDNA antibody),anti-small nuclear ribonucleoprotein antibody(anti-snRNP antibody)and serum total IgG and IgM in lupus mice.3.P2Y6 KO alleviates glomerular IgG and complement C3 deposition in lupus erythematosus mice.The proportion of inflammatory cells in abdominal cavity of mice was detected by flow cytometry 2 weeks after injection of Pristane.The results showed that P2Y6 KO significantly decreased the proportion of monocytes and surface activation/maturation markers of Ly6Chi in abdominal cavity of mice.5.The proportion of spleen lymphocytes of lupus mice was detected by flow cytometry 8 months after Pristane injection.The results showed that P2Y6 KO significantly decreased the proportion of CD4+T cells and the activation of T cells in the spleen of lupus mice.6.P2Y6 KO accelerated the transformation of bone marrow cells from Ly6Chi monocytes to Ly6Clow monocytes.Conclusion:This study shows that the expression in peripheral blood mononuclear cells of patients with SLE is up-regulated.The deletion of P2Y6 receptor significantly reduced the production of anti-double-stranded DNA antibody(anti-dsDNA antibody)and anti-small nuclear ribonucleoprotein antibody(anti-snRNP antibody)and the deposition of glomerular IgG and complement C3 in lupus mice induced by Pristane,thus alleviating the disease progression of lupus mice.This may be achieved by reducing the number of inflammatory monocytes in the early stage of the disease and by reducing the number of pathogenic CD4+T cells and affecting the terminal differentiation of B cells in the progressive stage of the disease.Figure 10 table 3 reference 66...