摘要
目的:观察腺病毒载体搭载的人转化生长因子基因对兔椎间盘退行性变模型的治疗作用。方法:实验于2003-03/2004-09在协和医院骨科实验室进行。取4个月龄健康雌性日本大白兔30只,于协和医院骨科实验室行L4～5,L5～6前外侧纤维环损伤手术以构建椎间盘退行性变模型。术后8周将动物随机分入3组。治疗组14只,注射20μL(10×106pfu)腺病毒载体介导人转化生长因子β1基因;对照组8只,注射20μL(10×106pfu)腺病毒载体介导标志基因(Ad/CMV-LacZ);空白组6只,注射20μL生理盐水。另取2只同类的健康白兔作为正常对照,不予任何手术或治疗。注射后3周取各组椎间盘组织行苏木精-伊红染色、T2加权磁共振成像、蛋白免疫印迹、转化生长因子β1免疫组织化学观察。结果:①兔椎间盘组织切片观察结果:注射3周后,治疗组切口已基本愈合,髓核细胞大量增生;空白组纤维环外侧内2/3部分仍未愈合,部分髓核细胞坏死、凋亡;对照组较空白组改善不明显。②MRI扫描结果:治疗组椎间盘信号较对照组及空白组有所恢复,但并未完全达到正常椎间盘水平。③蛋白质免疫印迹分析结果:治疗组转化生长因子β1含量较对照组和空白组有显著上升(P<0.01)。④转化生长因子β1免疫组织分析结果:治疗组转化生长因子β1为强阳性表达(>50%),空白组几乎没有转化生长因子β1表达,对照组与空白组相似。治疗组显著高于对照组(χ2=20.563,P=0.00)。结论:腺病毒载体介导人转化生长因子β1基因转染退行性变的椎间盘细胞能够有效提高髓核细胞内转化生长因子β1含量,阻逆椎间盘退行性变。
AIM： To observe the therapeutic effect of adenovirus/cytomegalovirus human transforming growth lector-β1 （Ad/CMV-hTGF-β1） on degeneration of intervertebral disc. METHODS： The experiment was carried out in the orthopedic lab between March 2003 and September 2004. Thirty healthy 4-month female Japanese white rabbits were made into models of intervertebral disc degeneration by means of L4-5,L5-6 anterior-lateral anulus fibrosus injury operation. Eight weeks after operation, the rabbits were randomly divided into 3 groups： treatment group [n=14, the rabbits were injected with 20μL （10×10^6pfu） Ad/CMV- hTGF-β1 gene], control group [n=8, the rabbits were injected with 20μL （10×10^6pfu） Ad/CMV-LacZ], blank group （n=6, the rabbits were injected with 20μL saline）. Another 2 similar healthy rabbits without any treatment were taken as normal control group. Three weeks after the injection, corresponding discs were collected for investigations of hematoxylin eosin staining （HE staining）, T2 weighted magnetic resonance image （T2 MRI）, Western blotting and immunohistochemical observation of TGF-131. RESULTS： ① Observational results of the intervertebral disc tissue section in rabbits： Three weeks after injection, the incisions in the treatment group almost healed, and there was plenty of hyperplasia of nucleus pulposus cells; In the blank group, 2/3 of the lateral anulus fibrosus still not healed, necrosis and apoptosis were observed in part of the nucleus pulposus cells; The ameliorations in the control group were not obvious as compared with those in the blank group. ② MRI results： The intervertebral disc signal in the treatment group was recovered as compared with those in the control group and blank group, but still not reached the normal level of intervertebral disc. ③ Western blotting analysis showed that the TGF-β1 content in the treatment group was significantly increased as compared with those in the control group and blank .group （P 〈 0.01）. ④ Immunohis tochemical results of TGF-β1 There was strong positive expression （〉 50%） of TGF-β1 in the treatment group, and nearly no TGF-β1 expression in the blank group, which was similar to that in the control group. It was signifi- cantly higher in the treatment group than in the control group （χ^2=20.563, P=0.00） CONCLUSION：Transfection of Ad/CMV- hTGF-β1 gene is a potential way to increase content of TGF-β1 and reverse the intervertebral disc degeneration.
出处
《中国临床康复》
CAS
CSCD
北大核心
2005年第34期99-101,i0005,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家教委资助课题
教外司留[2001(345)]~~
