CAJ | 학술논문

目的观察X射线照射后小鼠胸腺细胞中CD4+ CD25+ NRP1+ Treg细胞数量及TGF-β1分泌量的变化,并探索PLC/ PIP2信号通路在其中的作用机制.方法 36只ICR小鼠按随机数字表法分为假照组、0.5、1.0、2.0、4.0和6.0 Gy的不同剂量组,X射线深部治疗机进行全身照射,于照射后16 h应用流式细胞仪分析小鼠胸腺细胞中CD4+ CD25+ NRP1+ Treg细胞的变化,ELISA法检测胸腺细胞TGF-β1含量的变化.EL-4细胞株经PKC激动剂(PMA)、Ca2+阻断剂(TMB-8)作用2h后并经4 GyX射线照射,于照射后48 h应用流式细胞术及ELISA法分别检测CD4+CD25+ NRP1+ Treg细胞及TGF-β1含量的变化.结果小鼠胸腺细胞中NRP1+ Treg在0.5 GyX射线作用后稍有下降,于1.0 Gy降至最低(t=6.96,P＜0.01),而后呈剂量依赖性升高,于6.0 Gy升至最高(t =6.70,P＜0.01)；胸腺细胞TGF-β1在0.5 GyX射线照射后显著下降(t=12.53,P＜0.01),而1.0～4.0 Gy照射后则呈剂量依赖性升高,于6.0 Gy仍保持高水平(t=10.40 ～ 15.30,P＜0.01).PMA作用后,与假照组相比,单独PMA组NRP1+ Treg细胞表达显著降低(t=3.06,P＜0.01),而联合照射后表达则明显上调(t=8.27,P＜0.01),TGF-β1无论受照与否,其含量均明显降低(t=10.46 ～39.69,P＜0.01)；而TMB-8作用后,无论受照与否CD4+ CD25+ NRP1+ Treg的表达及TGF-β1的分泌量均显著上调(t=5.53～44.26,P＜0.01).结论高剂量电离辐射可以诱导小鼠胸腺细胞中NRP1+ Treg细胞表达并增加TGF-β1的含量,此现象可能与启动PLC/PIP2信号通路有关.
목적 관찰X사선조사후소서흉선세포중CD4+ CD25+ NRP1+ Treg세포수량급TGF-β1분비량적변화,병탐색PLC/ PIP2신호통로재기중적작용궤제.방법 36지ICR소서안수궤수자표법분위가조조、0.5、1.0、2.0、4.0화6.0 Gy적불동제량조,X사선심부치료궤진행전신조사,우조사후16 h응용류식세포의분석소서흉선세포중CD4+ CD25+ NRP1+ Treg세포적변화,ELISA법검측흉선세포TGF-β1함량적변화.EL-4세포주경PKC격동제(PMA)、Ca2+조단제(TMB-8)작용2h후병경4 GyX사선조사,우조사후48 h응용류식세포술급ELISA법분별검측CD4+CD25+ NRP1+ Treg세포급TGF-β1함량적변화.결과 소서흉선세포중NRP1+ Treg재0.5 GyX사선작용후초유하강,우1.0 Gy강지최저(t=6.96,P＜0.01),이후정제량의뢰성승고,우6.0 Gy승지최고(t =6.70,P＜0.01)；흉선세포TGF-β1재0.5 GyX사선조사후현저하강(t=12.53,P＜0.01),이1.0～4.0 Gy조사후칙정제량의뢰성승고,우6.0 Gy잉보지고수평(t=10.40 ～ 15.30,P＜0.01).PMA작용후,여가조조상비,단독PMA조NRP1+ Treg세포표체현저강저(t=3.06,P＜0.01),이연합조사후표체칙명현상조(t=8.27,P＜0.01),TGF-β1무론수조여부,기함량균명현강저(t=10.46 ～39.69,P＜0.01)；이TMB-8작용후,무론수조여부CD4+ CD25+ NRP1+ Treg적표체급TGF-β1적분비량균현저상조(t=5.53～44.26,P＜0.01).결론 고제량전리복사가이유도소서흉선세포중NRP1+ Treg세포표체병증가TGF-β1적함량,차현상가능여계동PLC/PIP2신호통로유관.
Objective To explore the role of PLC/PIP2 signal pathway in the changes of mouse thymus CD4 + CD25 + NRP1 + Treg and TGF-β1 after different doses of X-ray irradiation Methods 36 ICR mice were randomly divided into 6 groups according to the irradiation doses of 0,0.5,1.0,2.0,4.0 and 6.0 Gy,respectively.Flow cytometry was used to detect the expression of NRP1 + Treg,and ELISA was used to detect the expression of TGF-β1 in mouse thymocytes at 16 h post-irradiation.The EL-4 cells were irradiated by X-rays at the dose of 4.0 Gy after co-cultured with the PMA and TMB-8 for 2 hours.Flow cytometry was used to detect the expression of NRP1 + Treg,and ELISA was used to detect the changes of TGF-β1 at 48 h post-irradiation.Results The NRP1 + Treg appeared a transient decrease at both 0.5 and 1.0 Gy irradiation and reached its valley value at 1.0 Gy (t =6.96,P ＜ 0.01),then showed a dosedependent increase and reached its peak at 6.0 Gy (t =6.70,P ＜ 0.01).The TGF-β1 level decreased after 0.5 Gy X-rays (t =12.53,P ＜0.01),then increased at a dose-dependent manner and reached its peak at 4.0 Gy (t =10.40-19.56,P ＜ 0.01).Compared with the sham-irradiation,NRP1 + Treg was decreased significantly after PMA treatment (t =3.06,P ＜ 0.01),while it was up-regulated significantly after irradiation in the presence of PMA (t =8.27,P ＜ 0.01).TGF-β1 was reduced in the presence of PMA with or without irradiation (t =10.46-39.69,P ＜ 0.01).NRP1 + Treg and TGF-β1 were increased significantly after TMB-8 treatment (t =5.53-44.26,P ＜ 0.01).Conclusions NRP1 + Treg cells and TGF-β1 were up-regulated after a high dose radiation,and the PLC/PIP2 signal pathway may participate in the regulation.