• Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China. Corresponding author: ZHOU Jiansheng, E-mail: jpylc@byyfy.com.cn;
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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 α (TNF-α). 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-α, and DMEM/ F12 medium containing 10% PBS, 100 ng/ mL TNF-α, and Ad-hIGF-1 (multiplicity of infection of 50) were used in control group, TNF-α 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-α group had no expression. The cell apoptosis rates of TNF-α 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-α 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-α group, the cell apoptosis rates of Ad-hIGF-1 group and control group were significantly reduced (P  lt; 0.05); the cell apoptosis rate of Ad-hIGF-1 group was significantly higher than that of control group (P  lt; 0.05). Conclusion Ad-hIGF-1 could inhibit the apoptosis of nucleus pulposus cells induced by TNF-α.

Citation: ZHANG Changchun,CUI Guopeng,SHAO Chen,ZHOU Xinshe,XIAO Yuzhou,ZHOU Jiansheng.. EFFECTS OF RECOMBINANT ADENOVIRUS VECTOR CARRYING HUMAN INSULIN-LIKE GROWTH FACTOR 1 GENE ON THE APOPTOSIS OF NUCLEUS PULPOSUS CELLS IN VITRO. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(11): 1375-1379. doi: 10.7507/1002-1892.20130298 Copy

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