多药耐药基因1及其表达产物在激素耐药系统性红斑狼疮患者淋巴细胞上的表达  

The expression of multidrug resistance gene 1 and P-glycoprotein in peripheral blood lymphocytes of patients with steroid resistant systemic lupus erythematosus

作　　者：王梅英[1,2] 曾沛英[1] 周耿民[1] 吕基杨 朱乙声[1] 陈澄[1] 王庆文[1] 郭成山 

机构地区：[1]北京大学深圳医院风湿免疫科,518036 [2]深圳市宝安区人民医院风湿免疫科,518101

出　　处：《免疫学杂志》2017年第12期1053-1059,共7页Immunological Journal

基　　金：广东省医学科研基金(A2016296);深圳市科创委资助项目(JCYJ20160427190358849;JCYJ20170307112009204)

摘　　要：目的探讨多药耐药基因1(multidrug resistance gene,MDR1)及其表达产物P-糖蛋白(P-glycoprotein,P-gp)在激素耐药系统性红斑狼疮(systemic lupus erythematosus,SLE)患者淋巴细胞的表达及其意义。方法选取初发SLE患者60例,均给予激素和(或)免疫抑制剂治疗6个月,根据治疗后SLEDAI评分及症状改善情况,筛选出激素有效组(n=49)和激素耐药组(n=11),同时以健康体检者30例作为正常对照,RT-PCR检测各组患者外周血淋巴细胞MDR1 mRNA表达,流式细胞术检测P-gp的表达和罗丹明123(rhodamine123,R123)荧光流式细胞术检测P-gp的活性。以环磷酰胺(cyclophosphamide,CTX,100μmol/L)、霉酚酸酯(mycophenolic acid,MPA,100μmol/L)和P-gp特异性封闭剂(Tariquidar,100μmol/L)干预培养激素耐药组患者(n=11)外周血淋巴细胞,R123荧光流式细胞术检测各组P-gp的活性。结果激素耐药组MDR1 mRNA和P-gp的表达明显高于激素有效组(t=3.43,P=0.002)、(t=3.792,P=0.001)和对照组(t=4.54,P<0.001)、(t=4.631,P<0.001);激素耐药组P-gp活性明显高于激素有效组(t=3.062,P=0.019)和对照组(t=5.563,P<0.001)。CTX可以抑制淋巴细胞P-gp活性,MPA抑制P-gp活性的能力明显降低,两者比较差异有统计学意义(t=3.372,P=0.011)。结论 MDR1及其表达产物P-gp参与SLE激素耐药机制的形成,CTX可以通过抑制P-gp活性,在激素耐药SLE的治疗中发挥作用。This study was performed to investigate the expression of multidrug resistance gene 1（MDR1） andP-glycoprotein（P-gp） in peripheral lymphocytes from systemic lupus erythematosus patients with poor responses tosystemic steroid therapy.A total of 60 SLE patients who had received systemic steroid treatment for longer than 6 months and 30 healthy subjects were monitored.SLE patients were subclassified into those with steroid-effectivegroup（n=49） and steroid-resistant group（n=11） according to their disease activity（estimated by SLEDAI-2000）after treatment.RT-PCR was used to detect the expression of MDR1 mRNA,the flow cytometry was used to detectthe P-gp expression,and rhodamine 123 was used to detect the P-gp activity.Lymphocytes,obtained from 11 patients with steroid-resistant SLE,with high levels ofP-gp expression were treated with cyclophosphamide,mycophenolic acid or Tariquidar in vitro,and subjectedto measure Rh123-efflux activity.Data showed that,after 6 months of treatment,the expression of MDR1 mRNA and the expression and activity of P-pg from patients with steroid-resistant group were significantly higherthan steroid-effective group and healthy controls.Treatment of lymphocytes with 100 μmol/L cyclophosphamide or100 μmol/L Tariquidar in vitro induced up to a 2-fold increase in the mean fluorescence intensity,as detected bythe Rh123-efflux assay.In conclusion,the high expression levels of MDR1 mRNA and P-gp in the peripherallymphocytes of SLE patients lead to poor disease control by systemic steroids.CTX,as a special type ofP-Glycoprotein inhibitor,possesses a promising effect for overcoming P-gp-mediated steroid resistance byinhibiting the P-gp activity.

关 键 词：多药耐药基因 P-糖蛋白 系统性红斑狼疮 激素耐药 环磷酰胺 

分 类 号：R593.241[医药卫生—内科学]