The Research Of The Correlation Between Zinc Finger Protein ZBTB20 And B Cell Abnormality In The Pathogenesis Of SLE

Objective To investigate the expression of ZBTB20 in B lymphocytes of SLE patients and in lupus mice and its relationship with B cell differentiation-related transcription factor Blimp-1,and explore the correlation between ZBTB20 and the pathogenesis of SLE.Methods(1)Peripheral blood mononuclear cells(PBMCs)were isolated from 36 cases patients with newly diagnosed SLE and 30 cases normal controls.(2)CD19+B cells were separated from PBMCs and enriched by immunomagnetic beads method and the m RNA expression levels of ZBTB20,Blimp1 in B cells were detected by real-time fluorescence quantitative polymerase chain reaction.The other PBMCs were detected by flow cytometry(FCM)to detect the ratio of B cell subsets.(3)To analyze the correlation between the expression of ZBTB20 m RNA and the ratio of B cell subsets(including CD19+B cells and CD19-CD138+ plasma cells), the expression of Blimp1 m RNA expression,anti-ds-DNA antibody and immunoglobulin Ig G in SLE patients.(4)Lupus Model MRL / lpr mice,NZB / WF1 mice were cultured for 25-28 weeks with the most serious disease condition,and the same period C57 BL / 6 mice were used as controls,and we detect the levels of anti-ds-DNA Antibody and immunoglobulin Ig G in serum.(5)The spleen,lymph nodes and thymus of mice were removed,and some of them were used to prepare paraffin sections for immunohistochemistry to detect the positive expression of ZBTB20,and the other part was used to extract total RNA,the m RNA expression levels of ZBTB20,Blimp-1 was detected by real-time fluorescence quantitative polymerase chain reaction.Results(1)The expression of ZBTB20 m RNA in peripheral blood CD19 + B cells in SLE patients was significantly higher than that in normal controls [(1.106 ± 0.161)vs(2.527 ± 0.355),P <0.05)];and the expression of Blimp1 m RNA in SLE patients was also significantly higher than that in normal control group [(1.106 ± 0.161)vs(2.527 ± 0.355),P <0.05](2)The proportion of CD19 + B cells in peripheral blood of SLE patients was lower than that of control group([4.87 ± 3.49)vs(6.92 ± 3.85),P <0.05),while the ratio of CD19-CD138+plasma cells and plasma cells /B cells were significantly higher than that of the control group [(0.49±0.41)vs(0.27±0.25),P<0.05;(0.14±0.05)vs(0.03±0.03),P<0.05],the difference was statistically significant.(3)The expression of ZBTB20 m RNA in B cells was positive correlated with the ratio of CD19-CD138 + plasma cells and plasma cell / B cell [(r = 0.165,P = 0.028),(r = 0.225,P = 0.021)],and was positive correlated with the expression of Blimp1 m RNA(r = 0.112,P = 0.039)and was positive correlated with the levels of anti-dsDNA antibody and immunoglobulin Ig G [(r = 0.415,P = 0.007),(r = 0.507,P = 0.005)].(4)MRL / lpr mice,NZB / WF1 mice were in severe morbidity,the spleen,lymph nodes and thymus were enlarged,the levels of anti-ds-DNA antibodies and immunoglobulin Ig G were higher than that in control group [(5.27 ± 3.79)vs(0.71 ± 0.65),P <0.05;(4.50 ± 2.18)vs(0.71 ± 0.65),P <0.05),[(12.35 ± 7.74)vs(1.68 ± 0.75)Vs(1.68 ± 0.75),P <0.05).The difference was statistically significant,indicating that the lupus models meet the experimental expectations.(5)The expression of ZBTB20 m RNA in the spleen of MRL / lpr mice and NZB / WF1 mice were significantly higher than that of the control group(0.49 ± 0.41 vs 0.27 ± 0.25),P <0.05(0.14 ± 0.05)vs(0.03 ± 0.03),P <0.05).Compared with the control group,the expression of ZBTB20 m RNA in the lymph nodes and thymus of The lupus model groups have no statistical significance.The results of immunohistochemistry showed that the expression of ZBTB20 in the spleen of the lupus model MRL / lpr and NZB / WF1 mice were significantly higher than that of the control group(21.3% vs 5.2%),(40.2% vs 5.2%)in the spleen of mice(P <0.05).However,the positive expression rate of ZBTB20 in the lymph nodes and thymus were not statistically significant compared with the control group.Conclusions 1.The expression of ZBTB20 m RNA in peripheral blood CD19 + B cells of SLE patients was significantly higher than that of the control group,and was positively correlated with the ratio of CD19-CD138 + plasma cells,and autoantibodies level in peripheral blood,therefore,ZBTB20 may be related to the pathogenesis of SLE.2.The expression of ZBTB20 m RNA in SLE patients was positively correlated with the expression of Blimp1 m RNA,which was related to the abnormal differentiation of B cells,which indicates that ZBTB20 is involved in the pathogenesis of SLE.3.Lupus mouse model experiments further indicate that the expression of ZBTB20 is associated with abnormal activation of B cells and differentiation into plasma cells,and ZBTB20 may be involved in the pathogenesis of SLE through this mechanism.