Zhao Ming, Liang Gongping, Luo Shuangyan, Lu Qianjin
Department of Dermatology, Central South University, Changsha, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2012 May;37(5):463-8. doi: 10.3969/j.issn.1672-7347.2012.05.006.
To investigate the effect of total glucosides of peony (TGP) on expression and DNA methylation status of ITGAL gene (CD11a) in CD4(+) T cells from patients with systemic lupus erythematosus (SLE).
CD4(+) T cells were isolated by positive selection using CD4 beads. CD4(+) T cells were treated by TGP at 0, 62.5, 312.5 and 1562.5 mg/L for 48 h. The MTT method was used to assess cell viability; mRNA expression level was measured by realtime-PCR; protein level of CD11a was measured by flow cytometric analysis; DNA methylation status was assayed by bisulfite sequencing.
No significant change in cell viability was found in CD4(+) T cells among the different concentration groups (P>0.05). Compared with control, the mRNA and protein levels of ITGAL were down-regulated significantly in SLE CD4(+) T cells treated with TGP (1562.5 mg/L) (P< 0.01). Furthermore, the extent of DNA methylation of ITGAL promoter was increased in TGP (1562.5 mg/L) treated CD4(+) T cells compared with control group (P<0.01).
TGP can repress CD11a gene expression through enhancing DNA methylation of ITGAL promoter in CD4(+) T cells from patients with SLE. This observation represents a preliminary step in understanding the mechanism of TGP in SLE therapy.
探讨白芍总苷（TGP）对系统性红斑狼疮（SLE）患者CD4(+) T细胞中ITGAL基因（CD11a）表达及DNA甲基化状态的影响。
采用CD4磁珠阳性分选法分离CD4(+) T细胞。将CD4(+) T细胞分别用0、62.5、312.5和1562.5 mg/L的TGP处理48小时。采用MTT法评估细胞活力；通过实时定量PCR检测mRNA表达水平；采用流式细胞术分析检测CD11a蛋白水平；采用亚硫酸氢盐测序法检测DNA甲基化状态。
不同浓度组CD4(+) T细胞的细胞活力无明显变化（P>0.05）。与对照组相比，用TGP（1562.5 mg/L）处理的SLE CD4(+) T细胞中ITGAL的mRNA和蛋白水平显著下调（P<0.01）。此外，与对照组相比，用TGP（1562.5 mg/L）处理的CD4(+) T细胞中ITGAL启动子的DNA甲基化程度增加（P<0.01）。
TGP可通过增强SLE患者CD4(+) T细胞中ITGAL启动子的DNA甲基化来抑制CD11a基因表达。这一观察结果是理解TGP治疗SLE机制的初步步骤。
