Experimental therapeutic effect of combined cytokines on beagle dogs exposed to 4.5 Gy γ-rays

Objective To evaluate the effects of combined administration of recombinant human interleukin-11(rhIL-11),recombinant human G-CSF(rhG-CSF)and recombinant human interleukin-2 (rhIL-2)on acute radiation sickness(ARS)beagles.Methods Sixteen beagle were irradiated with 4.5 Gy60 Co γ-rays to establish ARS models,and were divided into irradiation control group,supportive care group and combined cytokines treatment group.After irradiation irradiation control group was given no treatment,the dogs in supportive care group received purely symptomatic treatment,while combined cytokines treatment group received rhIL-11 50μg/(kg·d)and rbG-CSF 10μg/(kg·d)subcutaneously(0-14 d)and rhIL-2 1×1 06 U/d(29-43 d)besides symptomatic treatment.Manifestation and characteristics of ARS beagles were observed,and the survival time were recorded.At last,post-mortem examination and histological examination were performed.Results All animals underwent nausea,diarrhea and fever.After irradiation,all animals in irradiation control group died in two weeks,and the mean survival time was 12.7 d,while only one died at 33 d in supportive care group.All dogs in combined cytokine group survived at 45 day after exposure,and their haematopoiesis and gastrointestinal tract were recovered.Conclusions Combination of rhIL-11 + rhG-CSF + rhIL-2 treatment could be significantly effective on ARS beagles irradiated by 4.5 Gy60 Co γ-rays,which could accelerate injured haemotopoiesis and intestinal tract recovery,increase the survival rate and improve the life quality of animals.     

Initial research of screening for the differentially expressed proteins in beagles irradiated with 4.5 Gy60 CO γ-rays by two-dimensional gel electrophoresis and mass spectrometry

Objective To explore the mechanisms of cytokines on acute radiation disease in irradiated beagles.Methods The sera of beagles irradiated with 4.5 Gy γ-rays with cytokines treatment was collected at different time points post irradiation.The two-dimensional gel electrophoresis(2-DE)was used to isolate and compare the differentially expressed proteins in sera.HD-MS was used to analyze the differentially expressed proteins with significance,and the amino acid sequences should be determined. Results High resolution 2-DE gel map was obtained.There were six differentially expressed proteins in sera of irradiated beagles at different time points.Four protein spots were successfully identified by MS.A significant spot was identified as serum amyloid A(SAA)by HD-MS,with relative molecular mass of 13 077 and isoelectfie point of 6.26.Expression of SAA was not found 1 d pre-irradiation and 36 d postirradiation,but increased slightly 1 d(0.2166)and significantly 14 d post-irradiation(0.4577). Conclusions The expression of serum amyloid A was consistent with the process of acute radiation injury,which might indicate the turnover of the disease.     

Clotting mechanism in beagles irradiated by 4.5 Gy γ-rays

Objective To explore the clotting mechanism in beagles irradiated by 4.5 Gy γ-rays after treatment with supportive care,or supportive care and combined cytokines.Methods Sixteen beagles were divided into irradiation control group,Supportive care group and combined cytokines treatment group.Platelet aggregation test,thrombelagtography (TEG) and the time measurement were analyzed in vitro.Results In irradiation group and supportive care group,the platelet aggregation rates in beagles were decreased markedly and the k value of TEG was increased 7 d post-irradiation,while those indexes in combined cytokines treatment group changed little.At 14 d post-irradiation,each parameter of TEG in irradiated group changed obviously.The values of r,k,r+k and M were elevated significantly,clotting time and the maximum coagulation time of thrombus delayed,the Ma value was decreased markedly,and the maximum elasticity amplitude of thrombus was diminished.All parameters in combined cytokines treatment group were better than those in supportive care group.The thrombin time was prolonged obviously in irradiated group 14 d post-irradiation,while the thrombin time was the longest at 2-3 weeks post irradiation in supportive care group and combined cytokines treatment group(P>0.05).Conclusions Cytokines could improve the platelet aggregation and the blood clotting functions of beagles suffering from acute radiation sickness.     

辐射损伤的药物防护与治疗研究进展

随着核能、电磁能等在军事、科技、医学及日常生活中的广泛应用,人们接触辐射的机会越来越多,当辐射剂量超过一定范围时,就会给人们造成辐射损伤。如何预防辐射损伤、做好辐射损伤的医学救治,一直是人们研究的重点。     

Therapeutic effects of combined cytokines on hematopoietic injuries induced by 4.5 Gy γ-rays irradiation in beagles

Objectivc To observe the therapeutic effects of combined cytokines on hematopoietic injuries induced by 4.5 Gy60 Co γ-rays irradiation in beagles,and to provide experimental evidences for the clinical treatment of extremely severe myeloid acute radiation sickness(ARS).Methods 16 beagles were given 4.5 Gy60 Co γ-rays total body irradiation,and then randomly assigned into irradiation control group,supportive care group and cytokines group.In addition to supportive care,recombinant human granulocyte colony-stimulating factor (rhG-CSF),recombinant human interleukin-11(rhIL-11)and recombinant human interleukin-2(rhIL-2)were administered subcutaneouly to dogs in cytokines group.Peripheral blood hemogram was examined once every two days.Bone marrow and peripheral blood were collected to proceed colony cultivation 4 d pre-irradiation and 1 and 45 d post-irradiation.Conventional histopathological sections of sternum were prepared to observe the histomorphology changes. Results After irradiation,the population of all kinds of cells in peripheral blood declined sharply.WBC nadir Was elevated(1.04×109/L,but 0.28×109/L and 0.68×109/L for the irradiation control group and the supportive care group separately),the duration of thrombocytopenia was shortened (24 days,but 33 days for the supportive care groug) and red blood cell counts were maintained in the range of normal values after cytokincs treatment in combination.The colony forming efficiency of haemopoietic stem cells(HSCs)in bone marrow and peripheral blood decreased obviously 1 d post irradiation,but recovered to the level of that before irradiation 45 d post irradiation after supportive care and cytokines treatment.Hematopoietic cells disappeared in bone marrow of animals in irradiation control group,but hematopoietic functions were recovered after cytokines were administrated.Conclusions RhG-CSF.rhIL-11 and rhIL-2 used in combination could elevate WBC nadir,accelerate the recovery of leukocytes,platelets and red blood cells and promote the proliferation,differentiation and maturity of HSPCs left in the body after 4.5 Gy γ-rays total body irradiation,eventually restore the hematopoietic function.Hence,combination of rhG-CSF,rhIL-11 and rhIL-2 could serve as better therapeutic strategy to treat extremely severe myeloid ARS.     

细胞因子组合治疗急性放射损伤犬的组织病理学观察

急性放射性损伤的主要病理变化不仅是骨髓造血功能受到严重抑制,外周血白细胞和血小板极度缺乏,由此引起感染和出血,同时,放射性损伤还累及消化、呼吸、泌尿和生殖等多个系统,导致体内多个系统器官的损伤"[1-2].研究发现,动物放射性损伤后体内多种细胞因子、化学增活素和细胞外基质表达水平增高,以保护受损脏器的损伤"[3-4].而采用细胞因子治疗急性放射性损伤能促进造血功能的恢复[5].     

细胞因子联合对4.5Gyγ射线照射比格犬造血损伤的治疗作用

目的 观察细胞因子组合对4.5 Gy γ射线照射比格犬造血系统损伤的治疗效果,为极重度骨髓型急性放射病的临床救治提供实验依据。方法 16只比格犬均给予4.5 Gy ⁶⁰Co γ射线全身照射,随机分为照射对照、综合对症和细胞因子3组。细胞因子组动物在综合对症支持治疗的基础上应用rhG-CSF、rhIL-11和rhIL-2联合治疗。2 d检测1次外周血象,分别于照射前4 d,照射后1和45 d收集骨髓和外周血进行造血细胞集落培养,制备胸骨病理切片观察组织形态学改变。结果 照射后各组动物外周血各类细胞数急剧下降,细胞因子联合治疗可提高白细胞最低值(1.04×10⁹/L,而照射对照组和综合对症组分别为0.28×10⁹/L和0.68×10⁹/L),缩短血小板减少持续时间(细胞因子组24 d,综合对症组33 d),使红细胞维持在正常值范围;照射后1 d骨髓及外周血中造血干细胞集落形成率明显下降,照射后45 d 2个治疗组造血干细胞集落数均恢复为照射前水平;照射对照组动物骨髓造血细胞完全消失,细胞因子治疗使得骨髓造血功能完全恢复,与照射前水平比较差异无统计学意义。结论 rhG-CSF、rhIL-11和rhIL-2联合应用可提高极期时外周血白细胞最低值,加速白细胞、血小板和红细胞恢复,促进4.5 Gy γ射线照射犬体内残留造血干/祖细胞的增殖、分化和成熟,从而加速造血功能的重建。rhG-CSF、 rhIL-11和rhIL-2不失为治疗极重度骨髓型急性放射病的有效措施。     

极重度骨髓型急性放射病犬造血恢复的实验观察

3.27～4.19Gy照射所致的重度骨髓型急性放射病犬,经恰当的综台治疗后可以治活,其造血功能可以恢复。同样,部分5.5～7.0Gy照射所致的极重变骨髓型急性放射病犬,也可治活,其造血功能 仍有恢复的可能性。     

rhIL-3对化疗恒河猴和体外人骨髓造血祖细胞的影响

ｒｈＩＬ┐３对化疗恒河猴和体外人骨髓造血祖细胞的影响柳晓兰邱丽玲罗庆良孙连生夏贞彪董波毛秉智军事医学科学院放射医学研究所北京１００８５０国外大量文献资料表明重组人白介素３（ｒｈＩＬ－３）在肿瘤大剂量化疗等病人引起的白血病、血小板减少性疾病的治疗中显示...     

血小板生成素对猴急性放射病的治疗

本实验观察了重组人血小板生成素(recombinant human thrombopoietin,rhTPO)10μg·kg~(-1)·d~(-1)×21对5.0Gy300kV X线(20cGy/min)照射所致猴急性放射病的治疗效果。治疗组照后第一周内,外周血小板轻度升高至105％—114％,第7天开始下降,第10天达最低值(45％);此后开始迅速回升,第14天达照前值水平,第21天达最高值(311％),第30天仍为263％。对照组照后前6天外周血小板波动在67％—95％,第7天开始迅速下降,第16天达最低值9％,仅为治疗组的1/17(P<0.001);此后开始缓慢回升,第30天仍未恢复到照前水平。外周血网织红细胞数两组动物照后均迅速下降,治疗组于照后第8天达最低值(11％),低于照前值50％的时间为8天,第12天开始迅速回升,第20天达最高值(402％);对照组动物下降程度更为迅速,第8天仅为照前值的9％,低于照前值50％的时间长达16天,第27天达治疗组的最高值水平。病程极期对照组动物均接受了2—3次血小板悬液或全血输注,而治疗组动物均不需要输血或输血小板悬液。以上结果表明,重组人血小板生成素可明显地促进照射动物造血功能的恢复,尤其是促进巨核细胞生成血小板的恢复更为明显。