Role of human insulin-like growth factor-1 gene on collagen type Ⅱ expression of degenerative intervertebral disk
- VernacularTitle:hIGF-1基因转染促进退变椎间盘Ⅱ型胶原的表达
- Author:
Zongqiang HUANG
;
Shangli LIU
;
Zhaomin ZHENG
- Publication Type:Journal Article
- Keywords:
intervertebral disk;
degeneration;
expression;
insulin-like growth factor-1
- From:
Orthopedic Journal of China
2006;0(05):-
- CountryChina
- Language:Chinese
-
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.
