title: [Effects of recombinant adenovirus vector carrying human insulin-like growth factor 1 gene on the apoptosis of nucleus pulposus cells in vitro].
abstract: OBJECTIVE
  To investigate the effects of human insulin-like growth factor 1 (hIGF-1) gene transfected by recombinant adenovirus vector (Ad-hIGF-1) on the apoptosis of rabbit nucleus pulposus cells induced by tumor necrosis factor a (TNF-alpha).
  
  
  METHODS
  The intervertebral disc nucleus pulposus were harvested from 8 healthy adult domestic rabbits (male or female, weighing 2.0-2.5 kg). The nucleus pulposus cells were isolated with collagenase II digestion and the passage 2 cells were cultured to logarithm growing period, and then they were divided into 3 groups according to culture condition: DMEM/ F12 medium containing 10% PBS, DMEM/F12 medium containing 10% PBS and 100 ng/mL TNF-alpha, and DMEM/F12 medium containing 10% PBS, 100 ng/mL TNF-alpha, and Ad-hIGF-1 (multiplicity of infection of 50) were used in control group, TNF-alpha group, and Ad-hIGF-1 group, respectively. The results of transfection by adenovirus vector carrying hIGF-1 gene were observed by fluorescent microscopy; the expression of hIGF-1 protein was detected by Western blot, hIGF-1 mRNA expression by RT-PCR, and the cell apoptosis rate by TUNEL and flow cytometry.
  
  
  RESULTS
  Green fluorescence was observed by fluorescent microscopy in Ad-hIGF-1 group, indicating that successful cell transfection. The expressions of hIGF-1 protein and mRNA were detected in Ad-hIGF-1 group by Western blot and RT-PCR, while the control group and TNF-alpha group had no expression. The cell apoptosis rates of TNF-alpha group, Ad-hIGF-1 group, and control group were 34.24% +/- 4.60%, 6.59% +/- 1.03%, and 0.40% +/- 0.15%, respectively. The early apoptosis rates of TNF-alpha group, Ad-hIGF-1 group, and control group were 22.16% +/- 2.69%, 5.03% +/- 0.96%, and 0.49% +/- 0.05%, respectively; the late cell apoptosis rates were 13.96% +/- 4.86%, 10.68% +/- 3.42%, and 0.29% +/- 0.06%, respectively. Compared with TNF-alpha group, the cell apoptosis rates of Ad-hIGF-1 group and control group were significantly reduced (P < 0.05); the cell apoptosis rate of Ad-hIGF-1 group was significantly higher than that of control group (P < 0.05).
  
  
  CONCLUSIONS
  Ad-hIGF-1 could ingibit the apoptosis of nucleus pulposus cells induced by TNF-alpha.
authors: Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.
authors: Changchun
authors: C
authors: Zhang
authors: Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.
authors: Guopeng
authors: G
authors: Cui
authors: Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.
authors: Chen
authors: C
authors: Shao
authors: Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.
authors: Xinshe
authors: X
authors: Zhou
authors: Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.
authors: Yuzhou
authors: Y
authors: Xiao
authors: Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.
authors: Jiansheng
authors: J
authors: Zhou
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics* / Q000378:metabolism; D007403:Intervertebral Disc / Q000166:cytology* / Q000378:metabolism; D008297:Male; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: C583557:IGF1 protein, human; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D007334:Insulin-Like Growth Factor I