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The Anti-apoptotic Effect of Polypeptide from Chlamys farreri (PCF) in UVB-Exposed HaCaT Cells Involves Inhibition of iNOS and TGF-β1

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Abstract

To investigate the molecular mechanisms of polypeptide from Chlamys farreri (PCF)’s anti-apoptotic effect, HaCaT cells were exposed to 20 mJ/CM2 UVB, with or without pretreatment of TGF-β1 antagonist SB431542, inducible nitric oxide synthase (iNOS) inhibitor S-methylisothiourea sulfate (SMT), nitric oxide scavenger carboxy-PTIO, or 1.42, 2.84, and 5.69 mM PCF, or iNOS transfection (without UVB exposure). Apoptosis was confirmed with Hoechst 33258 staining; RT-PCR and western blot were used to determine the expression levels of iNOS and TGF-β1 signaling pathway. Both UVB exposure and iNOS transfection-induced apoptosis in UVB-exposed HaCat cells, while PCF, SB431542, SMT, and carboxy-PTIO all inhibited UVB-induced apoptosis. TGF-β1, Smad4, and Smad7 mRNA levels were all altered, similarly, iNOS, TGF-β1, and pSmad2/3 protein levels were all altered in UVB-exposed HaCaT cells. In pretreated cells, SB431542, SMT, carboxy-PTIO, and 1.42–5.69 mM PCF all inhibited UVB-induced apoptosis. Moreover, PCF treatment inhibited the expression levels of iNOS, TGF-β1, pSmad2/3, and Smad4, while increased the expression level of Smad7. SB431542 did not significantly alter iNOS expression, while SMT and carboxy-PTIO significantly altered TGF-β1 signaling level. The anti-apoptotic effect of PCF in UVB-exposed HaCaT cells involves the inhibition of iNOS expression and subsequently inhibition of TGF-β1 signaling pathway.

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Acknowledgments

This study was funded by the National Natural Science Foundation of China (Grant No. 30471458) and the Natural Science Foundation of Shandong Province, China (Grant No. ZR2011HM046).

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Authors and Affiliations

  1. Department of Pharmacology, Qingdao University Medical College, Boya Building Room 422, 308 Ningxia Road, Qingdao, 266071, Shandong, China

    Yantao Han, Qixiao Jiang, Hui Gao, Jie Fan, Feng Zhong & Chunbo Wang

  2. Shandong Mental Health Center, 49 Wenhua East Road, Jinan, 250000, Shandong, China

    Jie Fan

  3. The Affiliated Hospital of Medical College Qingdao University, 16 Jiangsu Road, Qingdao, 266003, Shandong, China

    Zhi Wang

  4. Yellow Sea Fishery Research Institute, 106 Nanjing Road, Qingdao, 266071, Shandong, China

    Yuan Zheng

  5. Department of Biology, Qingdao University Medical College, 308 Ningxia Road, Qingdao, 266071, Shandong, China

    Zhuoqun Gong

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  1. Yantao Han
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Correspondence to Chunbo Wang.

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Yantao Han and Qixiao Jiang have contributed equally to this work.

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Han, Y., Jiang, Q., Gao, H. et al. The Anti-apoptotic Effect of Polypeptide from Chlamys farreri (PCF) in UVB-Exposed HaCaT Cells Involves Inhibition of iNOS and TGF-β1. Cell Biochem Biophys 71, 1105–1115 (2015). https://doi.org/10.1007/s12013-014-0315-8

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