转染BMP7基因对兔髓核细胞生物学行为的影响

目的观察转染重组BMP7腺病毒（Ad-BMP7）后,兔髓核细胞BMP7和Ⅱ型胶原的表达情况。方法⑴取20只新西兰大白兔,15只作为实验组,5只作为实验对照组。实验组椎间盘注射Ad-BMP7,实验对照组注射携带绿色荧光蛋白（GFP）基因的腺病毒（Ad-GFP）,另设空白对照组。⑵分别于注射后1、2、4周各处死5只实验组兔子,注射后1周处死全部实验对照组兔子。RT-PCR和免疫组织化学染色检测BMP7的表达情况,免疫组织化学染色检测Ⅱ型胶原的表达情况。结果⑴实验组的髓核细胞显示了BMP7染色阳性结果,1、2、4周组阳性染色率差异无统计学意义（P〉0.05）。⑵实验组的髓核组织间质Ⅱ型胶原呈阳性染色,1、2、4周的灰度值单因素方差分析,差异有高度统计学意义（P〈0.01）;SNK两两比较,第1周与第2、第4周差异有统计学意义（P〈0.05）,第2、第4周间差别无统计学意义（P〉0.05）。结论Ad-BMP7转染兔髓核细胞后,能获得长期表达,并能促进Ⅱ型胶原合成。

Effects of BMP7 Gene Transfection on Rabbit Nucleus Pulposus Cells in vivo

Objective To observe the expression of BMP7 and collagen type Ⅱ in rabbit nucleus pulposus cells after the transfection of BMP7 recombinant adenovirus.Methods （1）Twenty New Zealand white rabbits were chosen and 15 rabbits in the experimental group while 5 rabbits in the experimental control group. Ad-BMP7 and Ad-GFP were injected directly into rabbit nucleus pulposus of lumbar discs in the experimental group and the experimental control group respectively. The self control group was still kept for reference. （2） The expression of BMP7 was detected by RT-PCR and immunohisto- chemistry （1st,2nd and 4th week）.Collagen type Ⅱ was detected by immunohistochemistry.Results （1） BMP7 gene showed positive staining results effectively in the experimental group. There was no signifi cant diffrence among the groups of the 1st, 2nd and 4th week（P〉0.05）. （2）The positive staining of collagen type Ⅱ in rabbit nucleus pulposus interstitial was able to be detected in the experimental group. There was significant difference between the 1st week group and the 2nd week and the 4th week group by the analysis of variance （P〈0.05）. There was no significant difference between the 2nd week and the 4th week group （P〉0.05）.Conclusion The transfection of BMP7 gene in rabbit nucleus pulposus cells can lead to its long-term expression as well as the stimulation to the synthesis of collagen type Ⅱ.