外周血CD4^+T细胞组蛋白乙酰化在狼疮性肾炎中的作用

The histone acetylation of CD4^+T cells of peripheral blood in the lupus nephritis

目的研究狼疮性肾炎(LN)患者外周血CD4+T细胞组蛋白乙酰化水平,探究其在LN发病中的作用。方法 18例LN患者均来自该科病房,全部患者都达到美国风湿学会1997年推荐的系统性红斑狼疮(SLE)的分类诊断标准。满足以下条件为活动性的LN,蛋白尿大于0.5g/d,或活动性尿沉渣改变(血尿红细胞超过5个/Hp,或脓尿白细胞大于5个/Hp,或细胞管型大于1个/Hp),血肌酐升高大于1.2mg/L,排除感染、肾结石和其他原因引起的。非活动的LN:蛋白尿小于0.5g/L,非活动性的尿沉渣。将LN患者分为两组:非活动性(I组)8例、活动性的LN组(A组)10例,对照组(N组)8例。分别采集LN患者和N组的外周血50mL,密度梯度离心法(Ficoll法)分离出外周血中的单个核细胞,采用免疫磁珠技术分选出CD4+T细胞,并提取组蛋白,采用组蛋白H3/H4乙酰化检测试剂盒检测组蛋白H3/H4的乙酰化水平,分析组蛋白H3/H4乙酰化水平及与疾病的关系。结果 (1)与N组相比,A和I组LN患者外周血CD4+T细胞的组蛋白H3、H4均呈低乙酰化状态(P<0.01);(2)A组的组蛋白H4的乙酰化水平低于I组(P<0.01),而组蛋白H3乙酰化水平两组间差异无统计学意义(P>0.05)。(3)组蛋白H4的乙酰化水平与24h尿蛋白排泄量呈负相关(r=-0.661,P<0.05)。结论在LN患者中的CD4+T细胞组蛋白H3和组蛋白H4均呈低乙酰化状态可能参与了LN的发病,CD4+T细胞组蛋白H4的乙酰化水平可能与LN的活动性有关。

Objective To study the histone acetylation level of CD4＋ T cells in peripheral blood of lupus nephritis ,to explore the role of histone acetylation in pathogenesis of lupus nephritis .Methods According to Feng X ,Bernstein ,Wagner S J and other scholars′s classification criteria for LN ,those who met the following conditions considered for the activity of LN ：proteinuria 〉0 .5 g/d ,change or activity of urinary sediment （hematuria 〉 5 red blood cell /Hp ,or pyuria 〉 5 Hp white blood cells ,or 1 cell type/Hp） ,serum creatinine increased 〉1 .2 mg/L ,and the exclusion of infection ,kidney stones and other causes .Lupus nephritis pa‐tients were divided into inactive group （group I） 8 people ,active group （group A） 10 people .18 patients with LN and 8 normal con‐trols were collected peripheral blood 50 mL ,density gradient centrifugation method （Ficoll method） for separation of mononuclear cells in peripheral blood;CD4＋ T cell was analyzed by immunomagnetic beads ,extracted histone acetylation level and detected H3/H4 protein by the acetylation of histone H3/H4 kits and the relationship of histone H3/H4 acetylation with diseases was analyzed . Results First ,compared with group N ,the histone H3 and H4 of CD4＋ T cells in peripheral blood both in A and I of group LN pa‐tients showed low acetylation status （P〈0 .01）;Second ,the acetylation level of histone H4 in group A was lower than that in group I （P〈0 .01） ,histione H3 acetyl the level of the group of two groups had no statistical significant （P〉0 .05） .Third ,it was nega‐tively related to the acetylation level of histone H4 and 24 h urinary protein excretion（r= -0 .661 ,P〈0 .05） .Conclusion Histone H3 and histone H4 of the CD4＋ T cells showed low acetylation may be involved in the pathogenesis of lupus nephritis .The acetyla‐tion level of histone H4 in CD4＋ T cells may be related to the activity of the LN .