CAJ | 학술논문

目的探讨青蒿琥酯治疗对SLE患者外周血Olf-1/EBF相关锌指蛋白(OAZ)信号通路相关基因表达水平的影响.方法应用随机数字表将24只12周龄MRL/lpr狼疮鼠分成2组,即狼疮对照组与青蒿琥酯治疗组,另取8只BABL/C作为正常对照组.采集并分离治疗前与治疗后4、8周狼疮鼠PBMCs,从细胞中提取RNA,逆转录为cDNA,用实时荧光定量PCR技术分析不同分组OAZ信号通路相关基因OAZ、Id1-3 mRNA表达水平的变化,ELISA法测定上清液中IL-4、IFN-γ、B细胞活化因子(BAFF)、ANA浓度并分析这些因子分泌水平与OAZ基因表达的相关性.2组比较采用t检验,多个样本均数间的比较采用单因素方差分析,两两比较采用LSD-t检验.结果治疗后8周,治疗组OAZ、Id1、Id3mRNA的表达水平(△Ct分别为12.9±0.8、12.0±0.4、10.2±0.8)与狼疮对照组(△Ct分别为9.8±1.0、9.3±1.1、8.1±0.8)和正常对照组(△Ct分别为13.9±1.2、11.4±0.7、4.7±0.8、10.3±1.0)比较差异有统计学意义(F=7.46,P=0.008; F=6.37,P=0.032; F=5.63,P=0.042),且OAZ基因mRNA表达水平△△Ct值与Id1、Id3表达水平△△Ct值呈正相关(r=0.867,0.947;P均＜0.05);其血清IL-4、BAFF浓度,ANA的吸光度值均较治疗前显著降低,IFN-γ较治疗前显著增高(P＜0.05),OAZ基因mRNA表达水平△△Ct值与Th1/Th2比值呈正相关(r=0.976,P＜0.01),与ANA吸光度比值、BAFF浓度比呈负相关(r=-0.955,r=-0.937;P＜0.05).结论青蒿琥酯治疗SLE可降低患者外周血中OAZ及其下游基因表达,同时减低多种细胞因子及ANA水平,推测OAZ信号通路可能是青蒿琥酯治疗SLE有效机制之一.
목적 탐토청호호지치료대SLE환자외주혈Olf-1/EBF상관자지단백(OAZ)신호통로상관기인표체수평적영향.방법 응용수궤수자표장24지12주령MRL/lpr랑창서분성2조,즉랑창대조조여청호호지치료조,령취8지BABL/C작위정상대조조.채집병분리치료전여치료후4、8주랑창서PBMCs,종세포중제취RNA,역전록위cDNA,용실시형광정량PCR기술분석불동분조OAZ신호통로상관기인OAZ、Id1-3 mRNA표체수평적변화,ELISA법측정상청액중IL-4、IFN-γ、B세포활화인자(BAFF)、ANA농도병분석저사인자분비수평여OAZ기인표체적상관성.2조비교채용t검험,다개양본균수간적비교채용단인소방차분석,량량비교채용LSD-t검험.결과 치료후8주,치료조OAZ、Id1、Id3mRNA적표체수평(△Ct분별위12.9±0.8、12.0±0.4、10.2±0.8)여랑창대조조(△Ct분별위9.8±1.0、9.3±1.1、8.1±0.8)화정상대조조(△Ct분별위13.9±1.2、11.4±0.7、4.7±0.8、10.3±1.0)비교차이유통계학의의(F=7.46,P=0.008; F=6.37,P=0.032; F=5.63,P=0.042),차OAZ기인mRNA표체수평△△Ct치여Id1、Id3표체수평△△Ct치정정상관(r=0.867,0.947;P균＜0.05);기혈청IL-4、BAFF농도,ANA적흡광도치균교치료전현저강저,IFN-γ교치료전현저증고(P＜0.05),OAZ기인mRNA표체수평△△Ct치여Th1/Th2비치정정상관(r=0.976,P＜0.01),여ANA흡광도비치、BAFF농도비정부상관(r=-0.955,r=-0.937;P＜0.05).결론 청호호지치료SLE가강저환자외주혈중OAZ급기하유기인표체,동시감저다충세포인자급ANA수평,추측OAZ신호통로가능시청호호지치료SLE유효궤제지일.
Objective To investigate the in vivo effects of artesunate (ART) treatment on OAZ gene expressions in MRL/lpr lupus mice.Methods Twenty-four 12-week-old female MRL/lpr lupus mice were randomly divided into two groups by the random number table,i.e.,the lupus control group and the ART treatment group,12 in each group.Besides,another 8 BABL/C mice were recruited as the normal control group.Peripheral blood mononuclear cells (PBMC) were collected from lupus mice before treatment as well as 4 week and 8 week after treatment,and RNA was extracted and reverse transcripted to cDNA.mRNA expression levels of OAZ and Id1-3 were measured by using real-time polymerase chain reaction (PCR),and the data between the two groups were analyzed by t test,a plurality of samples were compared with the single factor analysis of variance.Serum levels of IFN-γ,IL-4,BAFF and ANA were tested by enzyme-linked immunosorbent assay (ELISA).Relationships of the gene expression levels with levels of cytokines and ANA were analyzed.Statistical analysis were uising t test,ANOVA and LSD-t test.Results mRNA expression levels of OAZ,Id1 and Id3 gene (△Ct:12.9±0.8,12.0±0.4,10.2±0.8) in lupus mice were significantly different from 8 weeks after the treatment comparing to those in the lupus control group (△Ct:9.8±1.0,9.3± 1.1,8.1±0.8) and the normal control group (△Ct:13.9±1.2,11.4±0.7,4.7±0.8,10.3±1.0)(F=7.46,P=0.008; F=6.37,P=0.032; F=5.63,P=0.042),and alteration of OAZ mRNA expression levels before and after the treatment were positively correlated with changes of Id1,Id3 levels (r=0.867,0.947; P＜0.05),and levels of IL-4 and ANA were significantly lower 8 weeks after the treatment than those in the lupus control group (P＜0.05); while level of IFN-γwas higher than those in the lupus control group (P＜0.05).Alteration of OAZ mRNA expression levels before and after the treatment were negatively correlated with changes of ANA,BAFF levels (r=-0.955,r=-0.937; P＜0.05) and positively correlated with changes of Th1/Th2 levels (r=0.976,P＜0.01).Conclusion The expression of genes involving in the OAZ and downstream gene are effectively reduced along with the alteration of several cytokines and ANA after ART treatment.OAZ signaling pathway can play an important role in ART treatment for SLE.