白细胞介素—11基因治疗促进放射损伤小鼠造血恢复的…

白细胞介素－１１（ＩＬ－１１）是一种新的造血生长因子，体内、外实验结果显示ＩＬ－１１具有显著的造血促进作用，它能作用于早期的造血干细胞及祖细胞，促进其增殖和分化。为了探讨ＩＬ－１１基因治疗放射损伤后造血恢复的促进作用，我们构建了含人ＩＬ－１１ｃＤＮＡ的逆转录病毒载体，并转染小鼠成纤维细胞系ＮＩＨ－３Ｔ３，经筛选获得了３株高表达ＩＬ－１１的转染细胞亚克隆，用高表达ＩＬ－１１的转染细胞作为载体细胞，进     

白细胞介素－3对放烧复合伤小鼠造血功能的影响

作者研究了ＩＬ－３对放烧复合伤动物骨髓造血功能的影响。发现３Ｔ３－ＩＬ－３细胞培养上清液中含有很高活性的ＩＬ－３。用它给放射损伤或放烧复合伤动物预防加治疗应用，对造血的刺激作用显著优于单独预防或单独治疗。对造血的这种良好作用既表现在造血实质细胞，也表现在造血基质细胞；在实质细胞中粒系祖细胞、红系早期和晚期祖细胞都有作用。ＩＬ－３对造血功能未表现抑制的烧伤动物未显示出对造血的刺激作用。     

重组人IL-11对照射小鼠造血系统的影响

目的：观察重组人白细胞介素－１１（ｒｅｃｏｍｂｉｎａｎｔ ｈｕｍａｎ ＩＬ－１１,ｒｈＩＬ－１１）对照射小鼠血恢复的影响。方法：给５．５Ｇｈ＾６０Ｃｏγ射照射小鼠连续１０ｄ皮下注射ｒｈＩＬ－１１ ２００,４０００μｇ／（ｋｇ．ｄ）或溶剂,观察体重、外周血象和髓髓造血祖细胞的变化。结果：各种给药组小鼠体重明显增加（Ｐ〈０．０１）。照后第１１天ｒｈＩＬ－１１ ２００μｇ／（ｋｇ．ｄ）组血小板计数检查值     

重组人白细胞介素-6对照射小鼠早期造血功能恢复的影响

作者报道了国内重组人白介素－６（ｒｈＩＬ－６）对６．５Ｇｙ照射Ｃｓ７ＢＬ／６Ｊ小鼠后１０天造血功能（主要是造血干细胞、粒系和红系祖细胞）恢复的影响．结果表明；ｒｈＩＬ－６对照后早期造血的影响与白细胞介素－６（ＩＬ－６）剂量有关，且作用是双向的．当ＩＬ－６剂量在２００～１０００μｇ·ｋｇ－１·ｄ－１时，脾重和内源性肿结节（ＣＦＵ－Ｓ）数随ＩＬ－６剂量增大而逐渐增加，与对照组相比，ＣＦＵ－Ｓ数的增加统计学意义更明显．与此同时骨髓有核细胞数和ＣＦＵ－ＧＭ产率以及ＣＦＵ－ＧＭ／股骨也明显增加，但１０００μｇ·ｋｇ－１·ｄ－１组的效果反而下降，因此，ｒｈＩＬ－６促进照后早期造血恢复的最佳剂量可能在５００μｇ·ｋｇ－１·ｄ－１左右．当ＩＬ－６剂量为１０μｇ·ｋｇ－１·ｄ－１时，对照射小鼠造血恢复起抑制作用。这些发现对ｒｈＩＬ－６的应用是很有实际意义的．     

rhIL-11对外周血小板数及造血祖细胞增殖的影响

观察rhIL-11在体外对人骨髓造血祖细胞集落形成和对3.0Gy照射猕猴的外周血小板数及骨髓造血祖细胞恢复的影响。结果显示，rhIL-11可明显提高照射猴外周血小板数的最低值，缩短血小板最低值的持续时间，加速血小板数的恢复，但未见到血小板数与rhIL-11剂量间的明显关系。rhIL-11不仅直接刺激CFU－MK增殖和分化，而且还可促进CFU－GM、CFU－e、BFU－e及早期的CFU－Mix的增殖。提示rhIL-11可能是治疗血小板减少和骨髓再生不良的一个有效药物。     

重组人白细胞介素—6对照射小鼠早期造血功能恢复的…

作者报道了国内重组人白介素－６（ｒｈＩＬ－６）对６、５Ｇｙ照射Ｃｓ７ＢＬ／６Ｊ小鼠后１０天造血功能（主要是造血干细胞、粒系和红系祖细胞）恢复的影响。结果表明：ｒｈＩＬ－６对照后早期造血的影响与白细胞介素－６（ＩＬ－６）剂量有关，且作用是双向的。当ＩＬ－６剂量在２００～１０００μｇ．ｋｇ＾－１．ｄ＾－１时，脾重和内源性脾结节（ＣＦＵ－Ｓ）数随ＩＬ－６剂量增大而逐渐增加，与对照组相比，ＣＦＵ－Ｓ数的增     

IL—11对造血的刺激作用及其在辐射损伤治疗中的应用

体内外实验结果表明，IL－11（白细胞介素－11），可刺激骨髓巨核素，粒系和红系造血祖细胞的增殖，升高外周血小板，白细胞和红细胞数。因此，IL－11作为一种新的刺激骨髓造血细胞增殖的细胞因子具有它的特殊价值，可用于平时辐射事故病人和核战时伤员的救治，也可用于癌症患放，化疗后造血功能低下的治疗，具有广阔的临床应用前景。     

白细胞介素-3对放烧复合伤小鼠造血功能的影响

作者研究了ＩＬ－３对放烧复合伤动物骨髓造血功能的影响。发现３Ｔ３－ＩＬ－３细胞培养上清液中含有很高活性的ＩＬ－３。用它给放射损伤或放烧复合烧动物预防加治疗应用，对造血的刺激作用显著优于单独预防或单独治疗。对造血的这种良好作用既表现在造血实质细胞，也表现在造血基质细胞；在实质细胞中粒系祖细胞、红系早期和晚期祖细胞都有作用。ＩＬ－３对造血功能未表现抑制的烧伤动物未显示出对造血的刺激作用。     

rhIL-11联合rhG-CSF和CTX动员小鼠外周造血干/祖细胞的实验研究

为研究rhIL 11联合rhG CSF和CTX对C5 7BL/ 6小鼠外周造血干细胞的动员作用 ,将 48只C5 7BL/ 6小鼠随机分为 4组 ,3组d0给予一次腹腔注射 2 0 0mg/kg的CTX ,其中 2组d1d15分别皮下注射rhIL 11、rhIL 11加rhG CSF ,观察不同时间外周血白细胞、血小板计数、CD34 细胞比例、CFU GM、CFU MK及CFU E的数量变化。结果发现 ,rhIL 11或联合rhG CSF可明显促进CTX骨髓抑制小鼠外周血血小板和白细胞的恢复、提高外周血CD34 细胞比例及三系造血细胞集落数量。证明rhIL 11具有外周血造血干细胞动员作用 ,而与rhG CSF和CTX联合动员效果更佳。     

重组人白细胞介素-11不良反应的临床研究

目的:观察重组人白细胞介素-11(rhIL-11)在临床应用中的不良反应.方法:选择经病理学确诊的住院恶性实体瘤46例,随机分为观察组22例及对照组24例;观察组化疗后24 h开始应用rhIL-11,对照组应用安慰药.结果:rhIL-11相关不良反应有注射部位反应、结膜充血、头晕、乏力,以及肌肉和(或)骨痛等,程度Ⅰ～Ⅲ度;发生rhlL-11相关毛细血管渗漏综合征死亡1例.结论:rhIL-11临床使用安全,但个别病例可发生严重不良反应.     

真皮多能干细胞促进大鼠急性放射损伤造血恢复的研究

目的 观察真皮多能干细胞移植对大鼠急性放射损伤后造血恢复的作用。方法 分离新生大鼠真皮多能干细胞体外培养至第10代用于研究，观察干细胞层体外对正常大鼠骨髓粒-巨噬系祖细胞和红系祖细胞集落生长的影响；进一步将真皮多能干细胞经尾静脉输入5Gyγ射线全身照射的大鼠体内，检测移植后骨髓有核细胞、粒-巨噬系祖细胞集落CFU—GM、红系祖细胞集落CFU—E和外周血白细胞数量、血红蛋白含量等指标的恢复情况，观察4周后动物的存活率。结果 真皮多能干细胞层体外可促进正常大鼠CFU—GM和CFU—E集落的生长；全身输入干细胞促进了5Gyγ射线全身照射动物的外周血白细胞和骨髓有核细胞、CFU-GM和CFU—E集落数的恢复。实验组动物4周后的存活率也明显高于对照组。结论 真皮多能干细胞具有一定支持造血的功能，可促进急性放射损伤大鼠的造血恢复，为造血损伤相关疾病的干细胞治疗提供新的细胞来源途径。     

成骨细胞促进急性放射损伤小鼠骨髓造血系统及血管恢复的研究

目的 探讨成骨细胞对放射损伤小鼠骨髓造血系统及血管恢复的影响.方法 取18只雄性BALB/c小鼠股骨制备成骨细胞,其余42只小鼠随机分成健康对照组、成骨细胞组和生理盐水组3组.健康对照组不做任何处理;成骨细胞组和生理盐水组小鼠于6.0 Gy 60 Co γ射线一次性全身均匀照射后,分别经尾静脉输入成骨细胞(2×106个/只)和等体积的生理盐水.于照射后第7、14和21天计数小鼠外周血细胞,骨髓单个核细胞并作骨髓组织学观察.采用流式细胞仪检测骨髓单个核细胞中CD34+细胞百分比,采用免疫组织化学方法检测小鼠骨髓微血管密度.结果 照射后第7、14和21天成骨细胞组小鼠外周血细胞和骨髓单个核细胞计数,骨髓单个核细胞中CD34+细胞百分比,骨髓组织造血面积及骨髓微血管密度均明显高于生理盐水组(t=2.46～64.51,P＜0.05).结论 成骨细胞能促进放射损伤小鼠骨髓造血系统及血管的恢复.

Abstract：

Objective To explore the effects of osteoblasts on the recovery of hematopoiesis and angiogenesis in acute irradiation injury mice.Methods The femurs of 18 male BALB/c mice were used to prepare the bone marrow osteoblasts, and the rest mice were divided into 3 groups as normal group, saline group and osteoblast group.The mice in normal group received no treatment, and the other two groups were received 6.0 Gy 60Co γ-ray irradiation.After irradiation each mouse of osteoblast group was administered with 2 × 106 osteoblasts through tail vein injection, and equal volume saline was given to each mouse of saline group by the same way.The following factors were measured at 7, 14, 21 d after irradiation, they were the counts of peripheral blood cells and bone marrow mononuclear cells ( BMMNC ) , the percentage of CD34 + cells in BMMNC, the histology changes and micro vascular density (MVD) of bone marrow tissue.Results The counts of peripheral blood cells, BMMNC and hematopoietic tissue area in osteoblast group were higher than those in saline group.The percentage of CD34 + cells in BMMNC and the MVD of bone marrow in osteoblast group were also higher than those in saline group at 7, 14, 21 d after irradiation ( t = 2.46 - 64.51, P ＜ 0.05 ).Conclusions Osteoblasts could significantly promote the recovery of hematopoiesis and angiogenesis in mice after acute irradiation injury.     

小鼠受γ射线全身照射后FL表达变化的研究

目的 FL(fit3 ligand)是一种作用于早期造血细胞的生长因子，为进一步研究FL水平与辐射所致造血损伤的相关性，观察了小鼠受γ射线全身照射后FL的表达变化。方法以^60Coγ射线全身一次性均匀照射所致骨髓型急性放射病小鼠为模型，应用ELISA、流式细胞术、Western blot法检测照射前后小鼠外周血、骨髓、胸腺与脾脏中FL的表达变化。结果 小鼠受2～10Gyγ射线照射后24h，血清中FL浓度即有增加，并且随时间推移FL水平逐渐升高；吸收剂量在2～6Gy之间时，FL的水平随剂量增加而增加。6Gyγ射线照射后24h，外周血白细胞膜表面FL的表达增加，但24h后细胞膜表面FL的表达没有升高。6Gyγ射线照射后24h，骨髓、胸腺和脾脏有核细胞膜表面FL的表达明显升高，总蛋白中FL的水平亦明显增加。结论 辐射后早期小鼠FL可溶性与膜结合型蛋白的水平都有升高，而且辐射后血清中FL的水平与受照剂量及照射后时间存在相关性。     

白介素—10基因疗法延长异体皮肤移植存活期及其机理初探

目的：探讨成纤维细胞介导的IL－10基因疗法对异体皮肤移植存活期的影响与可能的机理。方法：小鼠腹腔内埋植高分泌IL－10的成纤维细胞，并移植异体鼠皮肤，观察排斥时间，测定受体鼠的淋巴细胞增殖反应、IFN－γ、IL－2、NK活性。结果：成纤维细胞介导的IL－10基因疗法显著延长异体皮肤移植的存活期，并降低小鼠淋巴细胞的增殖反应，减少IFN－γ、IL－2的产生及降低NK活性。结论：IL－10基因疗法可延长小鼠异体皮肤移植存活期，其机理与抑制机体的免疫效应因子有关。     

rhIL-11对急性放射病小鼠骨髓细胞凋亡的影响

目的　研究rhIL 11对急性辐射损伤后骨髓造血细胞凋亡的影响。方法　应用原位末端标记、流式细胞术及琼脂糖凝胶电泳法分别在照射后不同时间观察了rhIL 11对 5 5Gy6 0 Coγ射线全身照射小鼠骨髓造血细胞凋亡的影响。应用免疫组织化学法观察了各组小鼠骨髓细胞Bcl 2和P5 3蛋白的表达变化。结果　①照射前应用rhIL 11可使照射后 6h小鼠骨髓造血细胞凋亡率下降 ,而照射后给药组照射后 12h凋亡率下降更加明显。②照射后 3d给药组P5 3蛋白表达均较照射对照组明显降低 (P <0 0 1) ,Bcl 2蛋白表达则明显升高 ;照射后 7d给药组Bcl 2升高仍明显。结论应用rhIL 11可有效抑制照射后小鼠骨髓造血细胞凋亡 ,有利于造血功能的恢复。rhIL 11对细胞凋亡的影响可能与Bcl 2和P5 3等蛋白的调节密切相关。     

重组人白细胞介素-11和姜黄素对中子照射致小鼠肠道损伤的治疗作用

Objective To explore the therapeutic effect of recombinant human interleukin(rhIL-11) and curcumin on jejunal damage in mice after neutron irradiation.Methods 140 male BALB/c mice were randomly divided into 4 groups:20 mice in healthy control group,60 mice in mere irradiation group,30 mice in IL-11 treatment group and 30 mice in curcumin treatment group.The mere irradiation group mice were wholly exposed to 3 Gy neutron irradiation.The treatment groups mice were intraperitoneally enterocoelia once a day for 5 d after irradiation.The mortality of the mice were observed.The mice in the control and mere irradiation groups were killed 6 h,1,3,and 6 d post-irradiation,respectively,and the mice of the 2 treatment groups were killed 3 and 6 d post-irradiation,respectively and the samples of jujunum were colleted.HE staining,argyrophilic of nucleaolar organizer regions staining,Feulgen staining,and image analysis were used to observe the pathology and levels of argyrophilic proteins and DNA.Results The mice in the mere irradiation group all died at 5 d post-irradiation,while 2 mice in the IL-11 treatment group and 3 in the curcumin group survived.Large area necrosis and exfoliation were found in the intestinal epithelial mucosa of the mere irradiated group mice since 6 h to 3 d after irradiation.Crypt cell regeneration was seen occasionally found 3 days later and much more 5 days later.Crypt cell regeneration was obviously found in the intestinal epithelial mucosa and lots of new villi were observed 5 d after irradiation in both treatment groups,however,the amounts of crypt cells and new villi of the curcumin treatment group were less than those of the IL-11 treatment group.The contents of AgNOR and DNA in the intestinal epithelial cells 5 days after irradiation of the 2 treatment groups were all significantly higher than those of the mere irradiation group (F = 0.015-0.035,all P ＜ 0.05) but without significant differences between them.Conclusions Jejunal damage in mice could be induced after 3 Gy neutron irradiation.rhIL-11 and curcumin might reduce the damage and promote the regeneration and repair of the intestinal epithelium.     

Gene expression and gene therapy imaging

The fast growing field of molecular imaging has achieved major advances in imaging gene expression, an important element of gene therapy. Gene expression imaging is based on specific probes or contrast agents that allow either direct or indirect spatio-temporal evaluation of gene expression. Direct evaluation is possible with, for example, contrast agents that bind directly to a specific target (e.g., receptor). Indirect evaluation may be achieved by using specific substrate probes for a target enzyme. The use of marker genes, also called reporter genes, is an essential element of MI approaches for gene expression in gene therapy. The marker gene may not have a therapeutic role itself, but by coupling the marker gene to a therapeutic gene, expression of the marker gene reports on the expression of the therapeutic gene. Nuclear medicine and optical approaches are highly sensitive (detection of probes in the picomolar range), whereas MRI and ultrasound imaging are less sensitive and require amplification techniques and/or accumulation of contrast agents in enlarged contrast particles. Recently developed MI techniques are particularly relevant for gene therapy. Amongst these are the possibility to track gene therapy vectors such as stem cells, and the techniques that allow spatiotemporal control of gene expression by non-invasive heating (with MRI guided focused ultrasound) and the use of temperature sensitive promoters.     

rhIL-11对中子照射小鼠肠损伤的治疗作用

目的研究重组人白介素11(rhIL-11)对中子照射小鼠小肠上皮损伤及细胞周期的影响。方法采用裂变中子照射小鼠模型，观察rhIL-11对照射后小鼠小肠上皮形态、隐窝细胞坏死和细胞增生的影响，用流式细胞仪检测小鼠小肠上皮细胞周期的变化。结果rhIL-11治疗和预防给药可通过促进小肠隐窝细胞增殖使肠损伤得到迅速修复。照射后小鼠小肠上皮细胞发生明显的G2期阻滞，rhIL-11治疗可明显提高S期细胞比例。结论rhIL-11对中子照射小鼠小肠损伤具有防护作用，并能够影响细胞周期的变化。