ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Lang-chuang-ding Decoction (, LCD) on the expression of DNA methylation of CD70 gene promoter in peripheral blood mononuclear cells (PBMCs) of females with systemic lupus erythematosus (SLE).</p><p><b>METHODS</b>PBMCs isolated from female patients with SLE or healthy donors were cultured and treated with LCD medicated serum or normal serum for 24 or 48 h. The mRNA expressions of CD70 gene in PBMCs were detected by reverse transcription polymerase chain reaction (PCR); the DNA methylation of the CD70 gene promoter region was detected by methylation-specific PCR.</p><p><b>RESULTS</b>After treated with medicated serum for 48 h, the mRNA expression levels of CD70 in PBMCs of SLE patients were signifificantly higher than those of healthy donors (P<0.05); the DNA methylation levels of CD70 promoter region in PBMCs of SLE patients treated with medicated serum for 48 h were signifificantly higher than those treated with fetal bovine serum (P<0.01).</p><p><b>CONCLUSION</b>LCD could inhibit CD70 gene expression in PBMCs of SLE patients by promoting the DNA methylation of CD70 gene promoter.</p>
Subject(s)
Adult , Female , Humans , CD27 Ligand , Genetics , Metabolism , DNA Methylation , Genetics , Drugs, Chinese Herbal , Pharmacology , Gene Expression Regulation , Leukocytes, Mononuclear , Metabolism , Lupus Erythematosus, Systemic , Drug Therapy , Genetics , Promoter Regions, Genetic , RNA, Messenger , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the expression of CD70 and the methylation level of CD70 promoter in immuno-related pancytopenia（IRP）patients, and explore the role of CD70 in the pathogenesis of IRP.</p><p><b>METHODS</b>Thirty- five IRP patients and fifteen healthy donors were enrolled in this study. Peripheral blood mononuclear cells were isolated from venous blood by density gradient centrifugation, and CD4(+) T cells were isolated by immunomagnetic beads. The mRNA level and the percentage of CD70 of CD4(+) T cells were measured by real- time quantitative polymerase chain reaction（RT- PCR）and flow cytometry respectively. Methylation- Specific PCR（MSP）was performed to determine the methylation level of CD70 promoter.</p><p><b>RESULTS</b>The percentage of CD70 expression on CD4(+) T cells of untreated IRP patients［（7.46±1.51）%］was significantly higher than that of recovered ones［（5.95±1.34）%］and normal controls［（1.83 ± 0.60）%］, and that of recovered IRP patients was significantly higher than of normal controls（P<0.05）. The relative expressions of CD70 mRNA in CD4(+) T cells were 2.314（0.200-6.084）, 1.021（0.135-3.434）, 0.353（0.008-2.258）in three groups respectively. The differences among untreated IRP patients, recovered IRP patients and normal controls were significant（P<0.05）. The CD70 promoter methylation level in CD4(+) T cells of all IRP patients was significantly lower than that of normal controls （P<0.05）. The expression of CD70 positively correlated to the ratio of CD5(+) B cells（r=0.533, P<0.01）.</p><p><b>CONCLUSION</b>The overexpression of CD70 may lead to immunologic disarrangement of patients, which may play important role in the pathogenesis of IRP.</p>