生长激素缺乏性侏儒症基因治疗前瞻性研究:重组人生长激素非病毒载体基因治疗体内实验安全性观察  

Prospective study on gene therapy for growth hormone deficiency dwarfism: in vivo experimental safety of gene therapy of recombinant human growth hormone non-viral vector

作　　者：杨孔宾[1] 胡志强[1] 王超[1] 董齐[1] 胡恩喜[1] 程玉[1] 初明[1] 

机构地区：[1]哈尔滨医科大学第一临床学院神经外科,黑龙江省哈尔滨市150001

出　　处：《中国临床康复》2004年第34期7698-7699,共2页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(39800050)~~

摘　　要：目的:研究携带重组人生长激素(hGH)基因非病毒载体pNMG3骨骼肌注射方法的急性期安全性。方法:实验于2001/2002在哈尔滨医科大学神经外科研究所和哈尔滨兽医研究所生物技术国家重点实验室进行。用纯化裸露质粒pNMG3向幼鼠胫前肌不同剂量单次注射,两周内不同时间RT-PCR检测转录,RI-A检测血清hGH蛋白表达,局部注射部位组织病检,测定血清中谷草转氨酶(GOT)和肌酐检测肝功、肾功。结果:30～480μgpNMG3剂量组肌肉注射两周后RT-PCR,RIA检测结果阳性;7.5～480.0μg剂量组两周后少数见脾组织细胞轻度至中度增多;各组GOT、肌酐无明显变化。120μg剂量组病理检查4h可见肌注部位明显白细胞浸润、脾组织细胞增生,3d时心肌炎性细胞浸润,以上变化随时间延长而减轻,两周后仅少数见脾组织细胞轻度至中度增多。结论:7.5～480.0μg剂量组肌肉注射是安全的;超急性期纯化裸露质粒肌肉注射局部肌肉、心肌、脾受累明显,但是可逆的。AIM:To study the safety of intramuscular injection of non viral vector pNMG3 of recombinant human growth hormone gene(hGH). METHODS:The experiment was conducted in the National Key Laboratory of Biological Technique,Harbin Institute of Veterinarian and Institute of Neurosurgery,Harbin Medical University between 2001 and 2002.A single different dose of purified naked plasmid pNMG3 was injected into rat anterior tibialis muscle.The expression of hGH gene was detected by RT PCR and radioimmunoassay(RIA),glutamic oxalacetic transaminase(GOT)and creatinine and histopathology test were used to evaluate the function of liver and kidney. RESULTS:Two weeks later,in the dose of 30 μg to 480 μg pNMG3,the results of RT PCR and RIA were both positive.Except for mild to moderate hyperplasia in the spleen in several cases,histological examination was normal,GOT and creatinine of these blood samples had no apparent difference in the dose of 7.5 to 480 μg pNMG3.Then,4 hours after the intramuscular injection of pNMG3 at a single dose of 120 μg,apparent white cells were found among the local injection sites,and lymphoid hyperplasia in the spleen was found,too.A small amount of lymphocytes among myocardia of several cases were observed 3 days after injection.These histological lesions decreased gradually. CONCLUSION:Intramuscular injection of pNMG3 at different single doses of 7.5 to 480 μg is safe.In the super acute period,local sites of injection,myocardia and spleen are impaired,but they are reversible.

关 键 词：生长激素缺乏 侏儒症 基因治疗 重组人生长激素 非病毒载体 实验 安全性 骨骼肌注射 

分 类 号：R584.21[医药卫生—内分泌] R394[医药卫生—内科学]