Öz
Objective: Peptidylarginine deiminases (PADs) are enzymes converting the arginine to citrulline. They play a role in embryogenesis and cell signaling activities. But excessive or dysregulated PAD levels were determined to be associated with disorders and to increase in many diseases. It has been shown that Chloramidine (Cl-amidine) used as a PAD inhibitor suppresses increased PAD activity and shows anti-cancer, anti-inflammatory and antioxidant activities. Anti-inflammatory and antioxidant properties play an important role in wound healing. In this study, the possible efficacy of Cl-amidine on wound healing in the keratinocyte cell line was investigated by considering these parameters.
Methods: Cell proliferation evaluations of Cl-amidine concentrations (500, 125, 31.25 and 7.81 µM) determined according to the results of MTT method on HaCaT keratinocyte cells were performed using Real-Time Cell Analysis System (RTCA DP). COL1A1 mRNA expression levels were analyzed by RT (Real Time)-PCR (Polymerase Chain Reaction) method at the concentrations where proliferation was achieved (125, 31.25 µM). Migration effects of Cl-amidine on cells were evaluated by performing scratch analysis. MTT results were statistically analyzed with one-way ANOVA and Tukey test, and p<0.05 was accepted as significant. RTCA DP and RT-PCR results were evaluated using device software programs.
Results: In the study, it was found that certain concentrations of Cl-amidine had a proliferative effect on HaCaT keratinocyte cells. It was determined that Cl-amidine increased the amount of type 1 collagen, which is an important parameter for wound healing, by RT-PCR method. In addition, according to scratch analysis, it was detected that it positively affected cell migration in relation to wound closure.
Conclusion: This research shows that Cl-amidine may have a significant potential for wound healing.
Anahtar Kelimeler
Peptidyl arginine deiminase (PAD) Cl-Amidine Wound healing HaCat keratinocyte cell line
Kaynakça
- Jones JE, Causey CP, Knuckley B, Slack-Noyes JL, Thompson PR. Protein arginine deiminase 4 (PAD4): Current understanding and future therapeutic potential. Curr Opin Drug Discov Devel. 2009; 12(5): 616-27.
- Chen CC, Isomoto H, Narumi Y, Sato K, Oishi Y, Kobayashi T, Yanagihara K, Mizuta Y, Kohno S, Tsukamoto K. Haplotypes of PADI4 susceptible to rheumatoid arthritis are also associated with ulcerative colitis in the Japanese population. Clin Immunol. 2008; 126: 165–171.
- Moscarello MA, Mastronardi FG, Wood DD. The role of citrullinated proteins suggests a novel mechanism in the pathogenesis of multiple sclerosis. Neurochem Res. 2007; 32: 251–256.
- Ishigami A, Ohsawa T, Hiratsuka M, Taguchi H, Kobayashi S, Saito Y, Murayama S, Asaga H, Toda T, Kimura N, Maruyama N. Abnormal accumulation of citrullinated proteins catalyzed by peptidylarginine deiminase in hippocampal extracts from patients with Alzheimer's disease. J Neurosci Res. 2005; 80: 120–128.
- Chang X, Han J, Pang L, Zhao Y, Yang Y, Shen Z. Increased PADI4 expression in blood and tissues of patients with malignant tumors. BMC Cancer. 2009; 9(40): 1-11.
- Vossenaar ER, Zendman AJ, van Venrooij WJ, Pruijn GJ. PAD, a growing family of citrullinating enzymes: genes, features and involvement in disease. Bioessays. 2003; 25(11): 1106-1118.
- Chang X, Han J. Expression of peptidylarginine deiminase type 4 (PAD4) in various tumors. Mol Carcinog. 2006; 45(3):183-96.
- Schellekens GA, de Jong BA, van den Hoogen FH, van de Putte LB, van Venrooij WJ. Citrulline is an essential constituent of antigenic determinants recognized by rheumatoid arthritis-specific autoantibodies. J Clin Invest. 1998; 101: 273–281.
- Luo Y, Arita K, Bhatia M, Knuckley B, Lee YH, Stallcup MR, Sato M, Thompson PR. Inhibitors and Inactivators of Protein Arginine Deiminase 4: Functional and Structural Characterization. Biochemistry, 2006; 45 (39): 11727–11736.
- Cau L, Takahara H, Thompson PR, Serre G, Méchin M, Simon M. Peptidylarginine Deiminase Inhibitor Cl-Amidine Attenuates Cornification and Interferes with the Regulation of Autophagy in Reconstructed Human Epidermis. J Invest Dermatol. 2019; 139(9): 1889-1897.
- Fredrikkson C. Keratinocytes in tissue engineering of human skin: in vitro and in vivo studies. Ġsveç: Linköping University. 2008.
- Shaw TJ, Martin P: Wound repair at a glance. J Cell Sci. 2009; 122: 3209-3213.
- Sgonc R, Gruber J. Age-related aspects of cutaneous wound healing: A mini-review. Gerontology. 2013; 59: 159-164.
- Carmicheal J, DeGrafit WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of tetrazolium-based semiautomated colorimetric assay: Assasment of chemosensivity testing. Cancer Research. 1987; 47: 936-942.
- Engür-Öztürk S, Dikmen M. Proteasome inhibitor immunotherapy for the epithelial to mesenchymal transition: assessing the A549 lung cancer cell microenvironment and the role of M1, M2a and M2c ‘hydrocortisone-polarised’macrophages. Molecular Biology Reports. 2022; 49(6): 4777-4793.
- Yuksel SN, Dikmen M, Canturk Z. Evaluation of Real Time Cell Proliferation, Anti-Inflammatory and Wound Healing Potential of Helenalin on HaCaT Keratinocytes Treated with Lipopolysaccharide Stimulated Monocytes. Indian Journal of Pharmaceutical Sciences. 2021; 83(2); 219-229.
- Safferling K, Sütterlin T, Westphal K, Ernst C, Breuhahn K, James M, Jäger D, Halama N, Grabe N. Wound healing revised: A novel reepithelialization mechanism revealed by in vitro and in silico models. Journal of Cell Biology. 2013; 203(4): 691-709.
- Evans ND, Oreffo ROC, Healy E, Thurner PJ, Man YH. Epithelial mechanobiology, skin wound healing, and the stem cell niche. Journal of the Mechanical Behaviour of Biomedical Materials. 2013; 28: 397-409.
- Diegelmann RF, Evans MC. Wound healing: an overview of acute, fibrotic and delayed healing. Frontiers in Bioscience. 2004; 9: 283-289.
- Sandt J, Roguet R, Cohen C, Esdaile D, Ponec M, Corsini E, Barker C, Fusenig N, Liebsch M, Benford D, Fraissinette AB, Fartasch M. The use of human keratinocytes and human skin models for predicting skin irritation. Alternatives to Laboratory Animals. 1999; 27: 723-743.
- Ranzato E, Patrone M, Burlando B. Platelet lysate stimulates wound repair of HaCaT keratinocytes. British Journal of Dermatology. 2008; 159: 537-545.
Öz
Kaynakça
- Jones JE, Causey CP, Knuckley B, Slack-Noyes JL, Thompson PR. Protein arginine deiminase 4 (PAD4): Current understanding and future therapeutic potential. Curr Opin Drug Discov Devel. 2009; 12(5): 616-27.
- Chen CC, Isomoto H, Narumi Y, Sato K, Oishi Y, Kobayashi T, Yanagihara K, Mizuta Y, Kohno S, Tsukamoto K. Haplotypes of PADI4 susceptible to rheumatoid arthritis are also associated with ulcerative colitis in the Japanese population. Clin Immunol. 2008; 126: 165–171.
- Moscarello MA, Mastronardi FG, Wood DD. The role of citrullinated proteins suggests a novel mechanism in the pathogenesis of multiple sclerosis. Neurochem Res. 2007; 32: 251–256.
- Ishigami A, Ohsawa T, Hiratsuka M, Taguchi H, Kobayashi S, Saito Y, Murayama S, Asaga H, Toda T, Kimura N, Maruyama N. Abnormal accumulation of citrullinated proteins catalyzed by peptidylarginine deiminase in hippocampal extracts from patients with Alzheimer's disease. J Neurosci Res. 2005; 80: 120–128.
- Chang X, Han J, Pang L, Zhao Y, Yang Y, Shen Z. Increased PADI4 expression in blood and tissues of patients with malignant tumors. BMC Cancer. 2009; 9(40): 1-11.
- Vossenaar ER, Zendman AJ, van Venrooij WJ, Pruijn GJ. PAD, a growing family of citrullinating enzymes: genes, features and involvement in disease. Bioessays. 2003; 25(11): 1106-1118.
- Chang X, Han J. Expression of peptidylarginine deiminase type 4 (PAD4) in various tumors. Mol Carcinog. 2006; 45(3):183-96.
- Schellekens GA, de Jong BA, van den Hoogen FH, van de Putte LB, van Venrooij WJ. Citrulline is an essential constituent of antigenic determinants recognized by rheumatoid arthritis-specific autoantibodies. J Clin Invest. 1998; 101: 273–281.
- Luo Y, Arita K, Bhatia M, Knuckley B, Lee YH, Stallcup MR, Sato M, Thompson PR. Inhibitors and Inactivators of Protein Arginine Deiminase 4: Functional and Structural Characterization. Biochemistry, 2006; 45 (39): 11727–11736.
- Cau L, Takahara H, Thompson PR, Serre G, Méchin M, Simon M. Peptidylarginine Deiminase Inhibitor Cl-Amidine Attenuates Cornification and Interferes with the Regulation of Autophagy in Reconstructed Human Epidermis. J Invest Dermatol. 2019; 139(9): 1889-1897.
- Fredrikkson C. Keratinocytes in tissue engineering of human skin: in vitro and in vivo studies. Ġsveç: Linköping University. 2008.
- Shaw TJ, Martin P: Wound repair at a glance. J Cell Sci. 2009; 122: 3209-3213.
- Sgonc R, Gruber J. Age-related aspects of cutaneous wound healing: A mini-review. Gerontology. 2013; 59: 159-164.
- Carmicheal J, DeGrafit WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of tetrazolium-based semiautomated colorimetric assay: Assasment of chemosensivity testing. Cancer Research. 1987; 47: 936-942.
- Engür-Öztürk S, Dikmen M. Proteasome inhibitor immunotherapy for the epithelial to mesenchymal transition: assessing the A549 lung cancer cell microenvironment and the role of M1, M2a and M2c ‘hydrocortisone-polarised’macrophages. Molecular Biology Reports. 2022; 49(6): 4777-4793.
- Yuksel SN, Dikmen M, Canturk Z. Evaluation of Real Time Cell Proliferation, Anti-Inflammatory and Wound Healing Potential of Helenalin on HaCaT Keratinocytes Treated with Lipopolysaccharide Stimulated Monocytes. Indian Journal of Pharmaceutical Sciences. 2021; 83(2); 219-229.
- Safferling K, Sütterlin T, Westphal K, Ernst C, Breuhahn K, James M, Jäger D, Halama N, Grabe N. Wound healing revised: A novel reepithelialization mechanism revealed by in vitro and in silico models. Journal of Cell Biology. 2013; 203(4): 691-709.
- Evans ND, Oreffo ROC, Healy E, Thurner PJ, Man YH. Epithelial mechanobiology, skin wound healing, and the stem cell niche. Journal of the Mechanical Behaviour of Biomedical Materials. 2013; 28: 397-409.
- Diegelmann RF, Evans MC. Wound healing: an overview of acute, fibrotic and delayed healing. Frontiers in Bioscience. 2004; 9: 283-289.
- Sandt J, Roguet R, Cohen C, Esdaile D, Ponec M, Corsini E, Barker C, Fusenig N, Liebsch M, Benford D, Fraissinette AB, Fartasch M. The use of human keratinocytes and human skin models for predicting skin irritation. Alternatives to Laboratory Animals. 1999; 27: 723-743.
- Ranzato E, Patrone M, Burlando B. Platelet lysate stimulates wound repair of HaCaT keratinocytes. British Journal of Dermatology. 2008; 159: 537-545.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Klinik Tıp Bilimleri (Diğer) |
| Bölüm | Araştırma Makaleleri |
| Yazarlar | |
| Yayımlanma Tarihi | 31 Ağustos 2023 |
| Yayımlandığı Sayı | Yıl 2023 Cilt: 9 Sayı: 3 |
