CAJ | 학술논문

目的观察再生障碍性贫血(AA)患者外周血单个核细胞(PBMNC)端粒长度及端粒酶活性与免疫抑制治疗(IST)疗效的关系,探讨AA发病机制及端粒长度对于评估IST疗效的意义.方法选取2010年9月至2013年3月71例AA患者,于初诊未接受治疗时留取外周血标本3 ml.采用流式-荧光原位杂交法(flow-FISH)检测PBMNC端粒长度,采用端粒重复序列扩增-PCR-酶联免疫吸附测定法检测端粒酶活性.结果各组PBMNC端粒长度均随患者年龄增长而缩短(b=-0.387,P=0.001).去掉年龄对PBMNC端粒长度影响,非重型AA(NSAA)组PBMNC端粒长度(30.957±4.502)与重型AA+极重型AA组(29.510±5.911)比较差异无统计学意义(P=0.573),均短于正常对照组(51.086±10.844)(P值均＜0.01).IST无效组患者初始PBMNC端粒长度(25.357±4.848)低于正常对照组(51.086±10.844),差异有统计学意义(P=0.005),而部分有效组(30.334±4.464)、完全有效组(32.808±4.685)与正常对照组比较差异均无统计学意义(P=0.517、P=0.254).当AA患者PBMNC端粒长度低于界值点(29.21％)时,IST无效的可能性更大.PBMNC端粒酶活性NSAA组(0.234±0.175)、SAA+VSAA组(0.324±0.178)均高于正常对照组(0.107±0.083),差异有统计学意义(P值均＜0.01).结论部分AA患者PBMNC端粒长度较同龄正常人缩短、端粒酶活性相对增高,此类患者IST可能无效,应尽早调整治疗方案.端粒或可成为预测AA患者IST疗效的指标.
목적 관찰재생장애성빈혈(AA)환자외주혈단개핵세포(PBMNC)단립장도급단립매활성여면역억제치료(IST)료효적관계,탐토AA발병궤제급단립장도대우평고IST료효적의의.방법 선취2010년9월지2013년3월71례AA환자,우초진미접수치료시류취외주혈표본3 ml.채용류식-형광원위잡교법(flow-FISH)검측PBMNC단립장도,채용단립중복서렬확증-PCR-매련면역흡부측정법검측단립매활성.결과 각조PBMNC단립장도균수환자년령증장이축단(b=-0.387,P=0.001).거도년령대PBMNC단립장도영향,비중형AA(NSAA)조PBMNC단립장도(30.957±4.502)여중형AA+겁중형AA조(29.510±5.911)비교차이무통계학의의(P=0.573),균단우정상대조조(51.086±10.844)(P치균＜0.01).IST무효조환자초시PBMNC단립장도(25.357±4.848)저우정상대조조(51.086±10.844),차이유통계학의의(P=0.005),이부분유효조(30.334±4.464)、완전유효조(32.808±4.685)여정상대조조비교차이균무통계학의의(P=0.517、P=0.254).당AA환자PBMNC단립장도저우계치점(29.21％)시,IST무효적가능성경대.PBMNC단립매활성NSAA조(0.234±0.175)、SAA+VSAA조(0.324±0.178)균고우정상대조조(0.107±0.083),차이유통계학의의(P치균＜0.01).결론 부분AA환자PBMNC단립장도교동령정상인축단、단립매활성상대증고,차류환자IST가능무효,응진조조정치료방안.단립혹가성위예측AA환자IST료효적지표.
Objective To observe the changes of telomere length and telomerase activity in patients with aplastic anemia (AA),and relationship with immunosuppressive therapy (IST) efficacy,to explore the pathogenesis of AA and the role of telomere length in evaluating immunosuppressive therapy efficacy.Method 71 cases of AA patients between September 2010 and March 2013 were enrolled into this study.3 ml peripheral blood specimens from this cohort of patients were collected to test the telomere length in peripheral blood mononuclear cell (PBMNC) with flow-FISH and detect telomerase activity with TRAP-PCR-ELISA method.Results Telomere length and age showed negative correlation (b=-0.387,P=0.001) in normal control,NSAA and SAA + VSAA groups,telomere length became shorter with the growth of age,and normal control group telomere length decreased along with the age growth slightly greater than the other two groups (NSAA,SAA+VSAA).Besides the effect of age on telomere length,(30.957±4.502) (29.510±5.911) no significant difference was observed between NSAA and SAA+VSAA groups (P=0.573),and NSAA,SAA+VSAA groups were significantly shorter than normal control group (51.086± 10.844) (P＜0.01).Telomere length in NR group (25.357±4.848) was significantly lower than normal control group (51.086± 10.844) (P=0.005),telomere length in CR (32.808±4.685)/PR groups (30.334±4.464) compared with normal control group had no significant difference (P=0.517,P=0.254).Telomere length below 29.21％ obviously decreased outcomes of IST.Telomerase activity had significant difference (x2=20.385,P＜0.01).The telomerase activity had no significant difference interms of age and gender in three groups,multiple comparison found that telomerase activities in SAA + VSAA (0.324± 0.178) (P＜0.01),and NSAA (0.234± 0.175)groups (P=0.002) were significantly higher than normal control group (0.107±0.083).Conclusion Telomere length of PBMNC in AA patients was significantly shortened than normal control group with telomerase activity increased,and telomere shorted more apparently in NR group,these patients should adjust the treatment as early as possible.Telomeres could predicte the curative effect of IST.