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[Effects of recombinant adenovirus vector carrying human insulin-like growth factor 1 gene on the apoptosis of nucleus pulposus cells in vitro].
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013 Nov; 27(11):1375-9.ZX

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).

CONCLUSION

Ad-hIGF-1 could ingibit the apoptosis of nucleus pulposus cells induced by TNF-alpha.

Authors+Show Affiliations

Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.Anhui Province Tissue Transplantation Key Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233004, P.R.China.

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

24501900

Citation

Zhang, Changchun, et al. "[Effects of Recombinant Adenovirus Vector Carrying Human Insulin-like Growth Factor 1 Gene On the Apoptosis of Nucleus Pulposus Cells in Vitro]." Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery, vol. 27, no. 11, 2013, pp. 1375-9.
Zhang C, Cui G, Shao C, et al. [Effects of recombinant adenovirus vector carrying human insulin-like growth factor 1 gene on the apoptosis of nucleus pulposus cells in vitro]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013;27(11):1375-9.
Zhang, C., Cui, G., Shao, C., Zhou, X., Xiao, Y., & Zhou, J. (2013). [Effects of recombinant adenovirus vector carrying human insulin-like growth factor 1 gene on the apoptosis of nucleus pulposus cells in vitro]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery, 27(11), 1375-9.
Zhang C, et al. [Effects of Recombinant Adenovirus Vector Carrying Human Insulin-like Growth Factor 1 Gene On the Apoptosis of Nucleus Pulposus Cells in Vitro]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013;27(11):1375-9. PubMed PMID: 24501900.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Effects of recombinant adenovirus vector carrying human insulin-like growth factor 1 gene on the apoptosis of nucleus pulposus cells in vitro]. AU - Zhang,Changchun, AU - Cui,Guopeng, AU - Shao,Chen, AU - Zhou,Xinshe, AU - Xiao,Yuzhou, AU - Zhou,Jiansheng, PY - 2014/2/8/entrez PY - 2014/2/8/pubmed PY - 2014/7/6/medline SP - 1375 EP - 9 JF - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery JO - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi VL - 27 IS - 11 N2 - 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). CONCLUSION: Ad-hIGF-1 could ingibit the apoptosis of nucleus pulposus cells induced by TNF-alpha. SN - 1002-1892 UR - https://www.unboundmedicine.com/medline/citation/24501900/[Effects_of_recombinant_adenovirus_vector_carrying_human_insulin_like_growth_factor_1_gene_on_the_apoptosis_of_nucleus_pulposus_cells_in_vitro]_ DB - PRIME DP - Unbound Medicine ER -