| BackgroundSystemic lupus erythematosus (SLE) is a polyclonal self-reactivematerial B cell-mediated autoimmune diseases, the direct causative factoris a polyclonal autoantibodies and the matter of immunocomplex formcause a variety of tissue and organ damage. For the exact cause of SLE isstill not very clear, but autoantibodies to the pathogenesis of SLE hasplayed a very important role has been a consensus. Clinical treatment ofSLE was the use of non-specific immunosuppressants, cytotoxic drugsand hormones, even though these drugs could effectively suppress theimmune cells, to control disease progression, but side effects were moreobvious, on the other hand, after treatment auto-reactive lymphocytespersists, it was difficult to achieve the purpose of healing. Althoughtreatment of SLE has been made great progress in recent years to produceantibodies to B lymphocytes in the field of differentiation into plasmacells, but this strategy can not effectively clear the plasma cells, longevityof plasma cells still secrete their own antibodies in vivo, these antibodiescan be activated again pathological immuneresponse, which led to theSLE disease recurrence, seeking to become the core issue has beenmarked a new target for plasma cells secrete antibodiesObjectiveThis study aimed to explore that whether promote B lymphocyte maturation protein1(Blimp1/PRDM1), B lymphocyte maturation antigen(BCMA) and the number of CD138+plasma cells were differences ofexpression in peripheral blood between patients with active systemic lupuserythematosus and healthy people, to reveal the relationship between theexpression of PRDM1, BCMA and the number of CD138+plasma cells.We contrasted SLE model mouse and normal mouse, to reveal the Blimp1expression in different tissues, as well as to reveal the relationshipbetween Blimp1and SLE at the same time. This study will clear Blimp1whether could become drug design to provide a new strategy for thetreatment of SLE, targeted to plasma cells, and lay the foundation for thefurther development of new drugs to treat SLE, which can also treat otherplasma cell diseases.Method1. We collected40patients with SLE blood samples for theexperimental group, and30healthy blood samples for the control group.Total RNA was extracted from peripheral blood mononuclear cells(PBMCs) and reversed transcriptase to obtaine cDNA, then we measuredby fluorescence quantitative PCR (FQ-PCR), calculated PRDM1BCMAgene expression according to the relative quantitative method.2. We collected40patients with SLE blood samples for theexperimental group, and30healthy blood samples for the control group.Three groups a) CD38-FITC, CD56-PE, CD45-PC5; b) CD19-FITC,CD138-PE, CD45-PC5; c) IgG1-PE/IgG1-FITC/IgG1-PC5, CD45-PC5to label cells, studyed the difference of the amount CD138+plasma cellsbetween experimental group and control group by Flow cytometry.3. SLE model mouse and normal mouse were killed at16weeks ofage under ether anesthesia, to observe the Blimp1protein expression andpathological changes in kidneys, spleen and lymph node tissue by immunohistochemistry, Western blot, HE staining. Extraction total RNAfrom the above-mentioned organizations, RT-PCR and gel electrophoresis,measuring the Blimp1gel optical density (OD) value, Calculating therelative multiples of Blimp1mRNA expression between model mice andnormal mice.Result1. We calculated△ct value of the SLE experimental group’s andcontrol group’s PRDM1and BCMA△ct values by SPSS statisticalsoftware and t-test, and found that there is statistically significant (P<0.05).The mean△ct of the experimental group’s PRDM1mRNA is0.67,BCMA mRNA is0.81; The mean△ct of the control group’s PRDM1mRNA was1.74, BCMA mRNA is1.72. This implied that SLEexperimental group’s PRDM1mRNA expression level was2.09timesthan the control group’s，and the BCMA expression level was1.87timesthan the control group’s by the Relatively quantitative method.2. Flow cytometry (FCM) analysied, the results suggest that therewere significant differences in the number of CD138+plasma cellsbetween SLE patients and control people in peripheral blood.3. The results showed that immunohistochemistry (IHC) opticaldensity（OD）value of Blimp1protein in kidney, lymph node and spleenof SLE model mouse, detected by Imagepro-Plus6.0Software, weresignificantly higher than in normal mice with statistical significance (P﹤0.05). The multiples were1.57,1.86and2.00in order. We analysisWestern blot’s and PCR products gel’s optical density (OD) values by thequantity one software (P﹤0.05), the calculation results showed that the expression of Blimp1protein in SLE model mouse’s kidneys, lymphnode and spleen tissues were1.54,1.99and2.21times than normalmouse’s; Blimp1mRNA expression in SLE model mouse’s kidneys,lymph nodes and spleen tissues were1.76,2.02and2.05times than thatof normal mouse. HE staining result showed that renal interstitial wasinfiltrated by inflammatory cells, focal plasma cells, lymphocytes, and asmall amount of macrophage cell infiltration. Occasionally finded thecells increase in glomerular; the basement membrane was slightlythickened. The folliculus lymphaticus hyperplasy, germinal centerexpansed. Splenic corpuscle’s bright and dark areas are normal structure,occasionally multinucleated giant cells in the red pulp obviouslyhyperplasy, A small amount of plasma cells around the small arteries.Conclusion1. We confirmed that PRDM1and BCMA mRNA were highexpression, and the increased number of CD138+plasma cells associatedwith the BCMA and PRDM1high expression. PRDM1and BCMA mRNAhighly expressed and stimulated the differentiation of plasma cells, mature,and secreted antibodies, and these antibodies produced a series ofpathological immune response, resulting in a variety of clinicalmanifestations.2. We initially confirmed that the Blimp1protein and Blimp1mRNAhighly expressed in SLE mouse’s kidney, lymph and spleen tissues; thepathological changed consistent with human SLE clinical features. Thiswould lay a good foundation for further research. |
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