1.ALKBH3-AS1 Expression in Peripheral Blood CD4+T Cells of Patients with Systemic Lupus Erythematosus and Its Correlation with Th17/Treg and Disease Activity
Yi QU ; Maoyuan WANG ; Youdong TANG ; Lihui HUNAG
Journal of Modern Laboratory Medicine 2024;39(5):107-111,212
Objective To investigate the expression of ALKBH3-AS1 in peripheral blood CD4+T cells of patients with systemic lupus erythematosus(SLE)and its correlation with T helper cell 17/regulatory T cells(Th17/Treg)and disease activity.Methods A total of 60 patients diagnosed with SLE in Leshan Hospital of Traditional Chinese Medicine from July 2020 to March 2024 were retrospectively collected.According to SLEDAI score,they were divided into active group(n=33,SLEDAI ≥ 10 score)and stable group(n=27,SLEDAI<10 score).At the same time,52 healthy subjects were selected as control group.The general data of three groups were collected and peripheral blood mononuclear cell(PBMC)was obtained by centrifugation in peripheral blood.CD4+T cells were isolated by immunomagnetic beads,and Th17/Treg ratio was detected by flow cytometry.The relative expression levels of ALKBH3-AS1 and retinoid-related orphan receptor γ t(ROR γ t)in CD4+T cells were detected by fluorescence quantitative PCR.The contents of transforming growth factor(TGF)-β and interleukin(IL)-17 in serum were determined by enzyme-linked immunosorbent assay(ELISA).The levels of C3 and C4 were determined by rate scattering immunoturbidimetry.Pearson analyzed the correlation between ALKBH3-AS1,Th17 and various clinical indicators in SLE patients.Logistic regression analysis of the influencing factors in patients with severe SLE showed that the difference was statistically significant.Results Hb,ALB,ALKBH3-AS1 mRNA,CD4+T,complement C3 and C4 in the control group were significantly higher than those in SLE group(t/Z=3.245,-11.169,-12.675,-17.829,-15.240,-19.212),RDW,TGF-β,RORγ t,Th17/Treg,IL-17,and CRP in the control group were lower than that in SLE group(t/Z=4.206,10.054,19.869,37.942,50.463,3.115),and the differences were statistically significant(all P<0.05).ALB,ALKBH3-AS1 mRNA,CD4+T in the active group were significantly lower than those in the stable group(t/Z=-8.918,-2.483,-11.694),CRP,TGF-β,RORγt,Th17/Treg,and IL-17 were significantly higher than those in the stable group(t/Z=3.121,5.671,1.787,14.720,12.044),and the differences were statistically significant(all P<0.05).Pearson analysis showed that ALKBH3-AS1 was positively correlated with CD4+T(r=0.663),and negatively correlated with Th17/Treg,IL-17,TGF-β,ROR γ t,SLED AIindex(r=-0.687,-0.715,-0.705,-0.678,-0.671),Th17/Treg was negatively correlated with CD4+T(r=-0.817),and positively correlated with IL-17,TGF-β,ROR γ t,SLED AI index with statistical significance(r=0.687,0.767,0.598,0.704).Logistics regression analysis,showed that increased CD4+T[OR(95%CI):0.715(0.304~0.904)]proportion and up-regulated ALKBH3-AS1[OR(95%CI):0.654(0.320~0.987)]expression were protective factors affecting disease activity in SLE patients.The contents of TGF-β[OR(95%CI):1.487(1.120~1.814)]and IL-17[OR(95%CI):1.294(1.217~1.887)]were up-regulated,the proportion of Th17/Treg[OR(95%)CI:1.674(1.361~1.679)]was up-regulated,and the relative expression of ROR γ t[OR(95%)CI:1.547(1.252~1.941)]was increased as risk factors affecting disease activity in SLE patients.Conclusion The down-regulated expression of ALKBH3-AS1 and up-regulated expression of TGF-β,ROR γ t and IL-17 in CD4+T cells of SLE patients are all correlated with disease activity,and can be potential biomarkers for diagnosis,disease activity and efficacy evaluation in SLE patients.