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| Cloning, Expression and Analysis of Antifibrosis Function of Fusion Protein Interleukin 10 with RGD |
| SHI Ji-hong, SHI Shan, BAI Xiao-zhi, GUAN Hao, TAO Ke, HU Xiao-long, LIU Jia-qi, HU Da-hai |
| Department of Burns and Cutaneous Surgery, Xijing Hospital, the Fourth Military Medical University, Xi'an 710032,China |
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Abstract Interleukin 10 (IL-10) is a pleiortropic cytokine which plays a pivotal role in the inflammatory response, immunological reaction and diseases development. RGD peptide has been demonstrated to bind to integrin on neovasculature endothelial cells specifically. A novel hybrid protein combining RGD with IL-10 was designed. The DNA sequence encoding recombination fusion protein IL10-RGD was subcloned into the pET22b(+) vector for protein expression in E. coli strain BL21(DE3). SDS-PAGE analysis showed an induced expression product band, with a molecular weight of about 19.3kDa, which was consistent with the expected value. The recombinant protein was isolated and purified by Sephadex G25M, and then renatured. Cultured human dermal fibroblasts were treated with transforming growth factor β1 (TGF-β1) stimulation and IL10-RGD treatment. The protein levels of various fibrosis related molecules were assessed by Western blot. The change of α-SMA positive fibroblasts was analyzed by immunocytochemistry. The results showed that IL10-RGD can significantly down-regulate the protein expression levels of Col1, Col3 and α-SMA, and decrease α-SMA positive cells in cultured dermal fibroblasts stimulation with TGF-β1. These findings indicate that IL10-RGD has the antifibrotic effect on dermal fibroblasts, the potential usage in fibrosis diseases by improvement the abnormal deposition of ECM, and suggest that IL10-RGD may be beneficial for the treatment of skin fibrosis and pathological scars.
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Received: 24 November 2011
Published: 25 February 2012
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Cite this article:
SHI Ji-hong, SHI Shan, BAI Xiao-zhi, GUAN Hao, TAO Ke, HU Xiao-long, LIU Jia-qi, HU Da-hai. Cloning, Expression and Analysis of Antifibrosis Function of Fusion Protein Interleukin 10 with RGD. China Biotechnology, 2012, 32(02): 17-23.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2012/V32/I02/17
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