作者：徐紅星, 邱德華,王慧娟 ,楊曉帆,季曉輝
【摘要】 目的 探討系統性紅斑狼瘡(SLE)患者凋亡調節蛋白受體Fas及共刺激分子CD40L的表達情況及其相互之間的調節。方法 採用酶聯免疫吸附試驗(ELISA)檢測SLE患者和正常人血清sFas和sCD40L的水平並分析兩者之間的相關性;同時分離10例SLE患者和10例正常對照的外周血淋巴細胞：①將細胞培養48 h後，採用流式細胞儀檢測T細胞亞群表面Fas及CD40L的表達情況;②在培養過程中分別加入抗FasL抗體阻斷Fas訊號及抗CD40L抗體阻斷CD40L訊號後，觀察T細胞亞群表面CD40L及Fas的表達。結果 ①SLE患者血清sFas和sCD40L平均水平分別為4.18 μg/L和5.87 μg/L，顯著高於正常對照組2.27 μg/L和2.31 μg/L,且SLE患者血清sFas活動期高於穩定期(P<0.01或P<0.05);sFas、sCD40L水平與SLEDAI(疾病活動指數)之間存在一定的正相關性(r=0.688,r=0.253，P均<0.05),但sFas與sCD40L水平之間未顯示明顯相關(r=0.201，P>0.05)。②經細胞培養後，SLE患者CD4(+)和CD8(+) T細胞表面Fas表達增加，與正常對照組比較，差異有顯著性(P<0.01或P<0.05);SLE患者CD4(+)和CD8(+) T細胞表面CD40L表達增加，與正常對照組比較，差異有顯著性(P均<0.05)。③SLE患者CD4(+)及CD8(+) T細胞上Fas與CD40L表達均呈正相關關係(r=0.311和r=0.517，P均<0.05)。④加入抗FasL抗體阻斷後，CD4(+)和CD8(+) T細胞表面CD40L表達下降，與阻斷前比較，差異有顯著性(P<0.05);加入抗CD40L抗體阻斷後，CD4(+)和CD8(+) T細胞表面Fas表達與阻斷前比較差異無顯著性(P>0.05)。結論 SLE患者血清sFas和sCD40L水平升高且呈正相關關係;T細胞亞群表面Fas及CD40L高表達且呈正相關關係;T細胞上Fas的表達調控CD40L的表達。說明凋亡調節蛋白Fas及共刺激分子CD40L可能共同參與SLE的發病過程。
【關鍵詞】 系統性紅斑狼瘡;Fas;CD40L
Abstract: Objective To investigate the expressions of Fas and CD40L in patients with systemic lupus erythematosus (SLE). Methods Serum sFas, sCD40L levels in patients with SLE and healthy controls were determined using a commercially available ELISA system and the correlation between the levels of sFas and sCD40L was analyzed. Meanwhile, peripheral blood mononuclear cells (PBMC) of patients with SLE and health controls were isolated by gradient centrifugation of heparinized blood. The expressions of Fas and CD40L on CD4(+) and CD8(+) T lymphocytes in patients with SLE and controls were determined by flow cytometry (FCM) after cultivation for 48 h. The antibodies of FasL and CD40L were add in the process of cultivation, and then the expressions of CD40L and Fas of peripheral blood T lymphocytes were determined. Results ①The average levels of sFas and sCD40L in SLE patients were 4.18 μg/L and 5.87 μg/L, respectively, which were markedly higher than those of 2.27 μg/L and 2.31 μg/L in the healthy controls; the serum levels of sFas and sCD40L were significantly higher in active SLE patients than those inactive patients (P<0.01, P<0.05); the serum levels of sFas and sCD40L were shown to have a positive correlation with SLEDAI scores (r=0.688, r=0.253,P<0.05), while no positive correlation was revealed between the levels of sFas and sCD40L (r=0.201,P>0.05). ②The expressions of Fas on CD4(+) and CD8(+) T lymphocytes in patients with SLE were significantly higher than those of the healthy controls (P<0.01 and P<0.05); the expressions of CD40L on CD4(+) and CD8(+) T lymphocytes in patients with SLE were significantly higher than that of the healthy controls (P<0.05). ③There was no correlations between the expressions of Fas on CD4(+) and CD8(+) T lymphocytes and those of CD40L (r=0.311, r=0.517). ④The expressions of CD40L on CD4(+) and CD8(+) T lymphocytes in patients with SLE significantly decreased after blockage by the antibody of FasL (P<0.05); there was no difference in the expression of Fas in patients with SLE after blockage by the antibody of CD40L (P>0.05). Conclusion The levels of sFas and sCD40L in SLE patients were higher than those of healthy controls and there was a positive correlation between them. The expressions of Fas and CD40L on CD4(+) and CD8(+) T lymphocytes in patients with SLE increased; and the expression of Fas had a positive correlation with that of CD40L; meanwhile, the expression of Fas regulated that of CD40L, which suggests that both Fas and CD40L are involed in the process of SLE.
Key words：systemic lupus erythematosus; Fas; CD40L