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Role of human insulin-like growth factor-1 gene on collagen type Ⅱ expression of degenerative intervertebral disk / 中国矫形外科杂志
Orthopedic Journal of China ; (24)2006.
Article in Chinese | WPRIM | ID: wpr-546042
ABSTRACT
[Objective]To study the role of hIGF-1 gene on collagen type Ⅱ expression of degenerative intervertebral disk.[Method]Twenty-four male New-Zealand rabbits intervertebral disk degenerontion(IVDD) models were done according to reference and randomly divided into Ad-CMV-hIGF-1,hIGF-1 growth factor and PBS group.Twenty five microlitre the second generation Ad/CMV-hIGF-1(T=80?109 PFU/L),hIGF-1 growth factor(100 ?g/L)and PBS were respectively injected into L4、5,L5、6 intervertebral disk under fluoroscopic guidance.One,two,four and eight weeks post-operation,rabbits were sacrificed,intervertebral disk samples were harvested.Total proteins of equal mass intervertebral disks were extracted,isolated in SDS-Polyacrylamide gel electrophoresis(SDS-PAGE)and transferred to polyvinylidene difluoride(PVDF)Millipore.The hIGF-1 growth factor expression were indentified with Western blot.Collagen type Ⅱ gene fragments were amplified with RT-PCR,and relative expression was done with GAPDH as intern control.[Result]The hIGF-1 interest protein existed at 7.6 Kilo-Dalton.At one week after injection,its expression quantities were almost equal between Ad/CMV-hIGF-1 and hIGF-1 growth factor group.At two week after injection,it obviously declined in hIGF-1 growth factor group.At four week after injection,it still expressed in Ad/CMV-hIGF-1 group.At eight week after injection,it did not express in theree groups.Collagen type Ⅱ mRNA relative expressions increased significantly from one to four weeks after injection,declined slightly at the end of eight weeks in Ad/CMV-hIGF-1 group.However,they appeared to decrease continuously in the other two groups with time.At the corresponding phases,those in PBS group were the lowest.[Conclusion]Ad/CMV-hIGF-1 could successfully infect degenerative intervertebral disk.The hIGF-1gene expression could last four weeks and could stimulate collagen type Ⅱ synthesis in Ad/CMV-hIGF-1 group.

Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Orthopedic Journal of China Year: 2006 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Orthopedic Journal of China Year: 2006 Type: Article