CAJ | 학술논문

目的探讨角质细胞生长因子(KGF)在白血病小鼠异基因脐带血移植(UCBT)中的作用及机制.方法 C57BL/6胎鼠外周血作为脐带血移植物.BALB/c小鼠随机数字表法分为7组,每组12只.对照1组:单纯接种白血病；对照2组:全身辐射(TBI)前第4天(-4d)接种白血病,单纯TBI处理；对照3组:不接种白血病,TBI处理后输注2×106个脐带血总有核细胞数(TNC)；对照4组:不接种白血病,自TBI-3 d起每日皮下注射磷酸盐缓冲液(PBS)连用7d,TBI后输注脐带血TNC,移植后第8天(+8 d)予输血小板支持；对照5组:TBI-4 d接种白血病,自TBI-3 d起每日皮下注射PBS连用7d,TBI后输注2×106个脐带血TNC,移植+8d予输血小板支持；实验1组:不接种白血病,自TBI-3 d起每日皮下注射KGF 1 mg/kg连用7d,TBI后输注脐带血TNC,移植+8d予输血小板支持；实验2组:接种白血病,自移植-3d起每日皮下注射KGF 1 mg/kg连用7d,TBI后输注脐带血TNC,移植+8d予输血小板支持.以生存期、病理组织学变化、脾脏淋巴细胞亚群、胸腺输出功能为观察指标并作组间比较.结果对照1组,12只小鼠全部死于白血病,小鼠生存时间为(11.1±1.5)d；对照2组,小鼠单纯TBI处理后12只全部死于造血衰竭,生存时间为(11.5±2.5)d；对照3组,5只(5/12)小鼠生存期超过100 d,7只小鼠20 d内死于内脏出血；对照5组,白血病小鼠脐带血移植后4只(4/12)生存期超过100 d；实验2组,白血病小鼠9只(9/12)生存期超过100d.实验2组与对照5组白血病小鼠生存时间比较差异有统计学意义(Log-rank检验,x2=4.996,P=0.0254).移植后对照4组小鼠脾脏T、NK和B细胞数分别为(9.32 ±0.48)×106、(1.59 ±0.11)×106、(18.74±2.01)×106个；实验1组小鼠脾脏T、NK和B细胞数分别为(13.20±1.14)×106、(1.75±0.12)×106、(20.36±0.86)×106个,实验1组T细胞、NK细胞数均高于对照4组(均P＜0.05).实验1组信号结合T细胞受体删除DNA环(sjTREC)水平明显高于对照4组[(228±24)拷贝/105脾细胞比(167±17)拷贝/105脾细胞,P =0.002].结论足月胎鼠外周血富含造血干祖细胞.KGF输注减少小鼠异基因脐带血移植后白血病复发,其机制为促进胸腺输出功能恢复. 目的探討角質細胞生長因子(KGF)在白血病小鼠異基因臍帶血移植(UCBT)中的作用及機製.方法 C57BL/6胎鼠外週血作為臍帶血移植物.BALB/c小鼠隨機數字錶法分為7組,每組12隻.對照1組:單純接種白血病；對照2組:全身輻射(TBI)前第4天(-4d)接種白血病,單純TBI處理；對照3組:不接種白血病,TBI處理後輸註2×106箇臍帶血總有覈細胞數(TNC)；對照4組:不接種白血病,自TBI-3 d起每日皮下註射燐痠鹽緩遲液(PBS)連用7d,TBI後輸註臍帶血TNC,移植後第8天(+8 d)予輸血小闆支持；對照5組:TBI-4 d接種白血病,自TBI-3 d起每日皮下註射PBS連用7d,TBI後輸註2×106箇臍帶血TNC,移植+8d予輸血小闆支持；實驗1組:不接種白血病,自TBI-3 d起每日皮下註射KGF 1 mg/kg連用7d,TBI後輸註臍帶血TNC,移植+8d予輸血小闆支持；實驗2組:接種白血病,自移植-3d起每日皮下註射KGF 1 mg/kg連用7d,TBI後輸註臍帶血TNC,移植+8d予輸血小闆支持.以生存期、病理組織學變化、脾髒淋巴細胞亞群、胸腺輸齣功能為觀察指標併作組間比較.結果對照1組,12隻小鼠全部死于白血病,小鼠生存時間為(11.1±1.5)d；對照2組,小鼠單純TBI處理後12隻全部死于造血衰竭,生存時間為(11.5±2.5)d；對照3組,5隻(5/12)小鼠生存期超過100 d,7隻小鼠20 d內死于內髒齣血；對照5組,白血病小鼠臍帶血移植後4隻(4/12)生存期超過100 d；實驗2組,白血病小鼠9隻(9/12)生存期超過100d.實驗2組與對照5組白血病小鼠生存時間比較差異有統計學意義(Log-rank檢驗,x2=4.996,P=0.0254).移植後對照4組小鼠脾髒T、NK和B細胞數分彆為(9.32 ±0.48)×106、(1.59 ±0.11)×106、(18.74±2.01)×106箇；實驗1組小鼠脾髒T、NK和B細胞數分彆為(13.20±1.14)×106、(1.75±0.12)×106、(20.36±0.86)×106箇,實驗1組T細胞、NK細胞數均高于對照4組(均P＜0.05).實驗1組信號結閤T細胞受體刪除DNA環(sjTREC)水平明顯高于對照4組[(228±24)拷貝/105脾細胞比(167±17)拷貝/105脾細胞,P =0.002].結論足月胎鼠外週血富含造血榦祖細胞.KGF輸註減少小鼠異基因臍帶血移植後白血病複髮,其機製為促進胸腺輸齣功能恢複.
목적 탐토각질세포생장인자(KGF)재백혈병소서이기인제대혈이식(UCBT)중적작용급궤제.방법 C57BL/6태서외주혈작위제대혈이식물.BALB/c소서수궤수자표법분위7조,매조12지.대조1조:단순접충백혈병；대조2조:전신복사(TBI)전제4천(-4d)접충백혈병,단순TBI처리；대조3조:불접충백혈병,TBI처리후수주2×106개제대혈총유핵세포수(TNC)；대조4조:불접충백혈병,자TBI-3 d기매일피하주사린산염완충액(PBS)련용7d,TBI후수주제대혈TNC,이식후제8천(+8 d)여수혈소판지지；대조5조:TBI-4 d접충백혈병,자TBI-3 d기매일피하주사PBS련용7d,TBI후수주2×106개제대혈TNC,이식+8d여수혈소판지지；실험1조:불접충백혈병,자TBI-3 d기매일피하주사KGF 1 mg/kg련용7d,TBI후수주제대혈TNC,이식+8d여수혈소판지지；실험2조:접충백혈병,자이식-3d기매일피하주사KGF 1 mg/kg련용7d,TBI후수주제대혈TNC,이식+8d여수혈소판지지.이생존기、병리조직학변화、비장림파세포아군、흉선수출공능위관찰지표병작조간비교.결과 대조1조,12지소서전부사우백혈병,소서생존시간위(11.1±1.5)d；대조2조,소서단순TBI처리후12지전부사우조혈쇠갈,생존시간위(11.5±2.5)d；대조3조,5지(5/12)소서생존기초과100 d,7지소서20 d내사우내장출혈；대조5조,백혈병소서제대혈이식후4지(4/12)생존기초과100 d；실험2조,백혈병소서9지(9/12)생존기초과100d.실험2조여대조5조백혈병소서생존시간비교차이유통계학의의(Log-rank검험,x2=4.996,P=0.0254).이식후대조4조소서비장T、NK화B세포수분별위(9.32 ±0.48)×106、(1.59 ±0.11)×106、(18.74±2.01)×106개；실험1조소서비장T、NK화B세포수분별위(13.20±1.14)×106、(1.75±0.12)×106、(20.36±0.86)×106개,실험1조T세포、NK세포수균고우대조4조(균P＜0.05).실험1조신호결합T세포수체산제DNA배(sjTREC)수평명현고우대조4조[(228±24)고패/105비세포비(167±17)고패/105비세포,P =0.002].결론 족월태서외주혈부함조혈간조세포.KGF수주감소소서이기인제대혈이식후백혈병복발,기궤제위촉진흉선수출공능회복.
Objective To explore the actions of keratinocyte growth factor(KGF)in leukemic mice allogeneic umbilical cord blood cell transplantation(UCBT)and elucidate its mechanism.Methods Peripheral blood drawn from the litters of C57BL/6 females was used as umbilical cord blood(UCB)graft.BALB/c mice were randomly divided into 7 groups(n =12 each).The grouping was as follows.Control group 1,inoculated with leukemia.Control group 2,inoculated with leukemia at-4 d and total body irradiation(TBI)treatment.Control group 3,TBI treatment and reconstituted with 2 × 106 UCB-TNCs.Control group 4,injected with PBS subcutaneously,TBI treatment and reconstituted with UCB-TNCs with platelet transfusion.Control group 5,inoculated with leukemia,injected with PBS subcutaneously,TBI treatment and reconstituted with UCB-TNCs with platelet transfusion.Experiment group 1,injected with KGF subcutaneously,TBI treatment and reconstituted with UCB-TNCs with platelet transfusion.Experiment group 2,inoculated with leukemia,injected with KGF subcutaneously,TBI treatment and reconstituted with UCB-TNCs with platelet transfusion.The survival status,pathohistological changes,splenic lymphoid cell subsets and thymic output post-UCBT were compared between groups.Results The survival time of control group 1 was(11.1 ± 1.5)days and all died of leukemia.The survival time of control group 2 was(11.5 ±2.5)days and all died of aplasia.Five of 12 mice of control group 3 survived for 100 days and 7 mice died of visceral hemorrhage.Four of 12 mice of control group 5 survived for 100 days and 8 mice died of leukemia with a survival rate of 33.3％.Nine of 12 mice of experiment group 2 survived for 100 days and 3 mice died of leukemia with a survival rate of 75.0％.The survival was prolonged in experiment group 2 mice as compared with that of control group 5 mice(x2 =4.996,P =0.0254).The splenic T,NK and B cell counts in control group 4 mice at +35 d were(9.32 ±0.48)× 106,(1.59 ±0.11)× 106 and(18.74 ±2.01)× 106 respectively.While in group 6 mice at + 35 d were(13.20 ± 1.14)× 106,(1.75 ± 0.12)×106 and(20.36 ±±0.86)× 106 respectively.The counts of T cell and NK cell of group 6 were higher than those of group 4(both P ＜ 0.05).The level of signal joint T-Cell receptor excision circles(sjTRECs)in control group 4 mice was(167 ± 17)copies per l05 cells while that of experiment group 1 mice(228 ±24)copies per 105 cells.They were higher than that of control mice(P =0.002).Conclusion Hematopoietic stem/precursor cells are abundant in full-term murine fetal peripheral blood.The infusion of KGF reduces the post-UCBT relapse of leukemia through the enhancement of thymic output.