Araştırma Makalesi
BibTex RIS Kaynak Göster

Prevalence of FV, FXIII, ACE, ApoE Gene Variants and Effects on Coronary Artery Disease Profile in Giresun

Yıl 2019, Sayı: 9, 864 - 880, 31.12.2019
https://doi.org/10.38079/igusabder.590895

Öz

Aim: Atherosclerosis-induced Coronary Artery Disease (CAD) is a multifactorial and polygenic complex disease. In recent studies, some genetic variants associated with CAD have come to the fore. In our study, four gene regions were analyzed for blood coagulation and lipid metabolism in order to determine the genetic profile which may be susceptible to CAD patients in Turkey (in living Giresun). These gene regions are: FV-Leiden (rs6025) (FVL), FXIII 163G> T (V34L) (rs: 5985), ACE (Angiotensin-converting enzyme) (rs1799752 I/D Polymorphism), ApoE rs429358).

Materials and Methods: In this study, 89 control and 174 patients were used. ApoE genotypes were determined by real time polymerase chain reaction (RT-PCR). FVL, FXIII, ACE were determined by PCR method.

Results: ACE D and ApoE e4 allele frequencies were lower in the control group (p = 0.026 and p = 0.015, respectively). Serum total cholesterol (p = 0.038) and LDL-K (p = 0.054) levels were higher in patients with FV A allele compared to those with GG genotype in the patient group. The presence of FXIII L allele was 31.9% in patients without MI. The presence of L allele shows a protective effect against the risk of MI (p = 0.06, OR = 0.464, 95% CI = 0.202-1.09).

Conclusion: None of the gene variants investigated in our study directly correlated with CAD development. However, the inferences we have obtained from our study, in which the gene variants investigated can indirectly affect CAD risk factors.

Kaynakça

  • Mozaffarian D, Benjamin EJ, Go AS, et al. American Heart Association statistics committee; stroke statistics subcommittee. Executive summary: heart disease and stroke statistics-2016 update: A report from the American Heart Association. Circulation. 2016;133(4):447-454.
  • Stranger BE, Forrest MS, Dunning M, et al. Relative impact of nucleotide and copy number variation on gene expression phenotypes. Science. 2007;315(5813):848-853.
  • Peden JF, Farrall, M. Thirty-five common variants for coronary artery disease: the fruits of much collaborative labour. Hum. Mol. Genet. 2011;20(R2):R198-205.
  • Wray NR, Goddard ME. Multi-locus models of genetic risk of disease. Genome Med. 2010;2(2):10
  • Consortium IKC. Large-scale gene-centric analysis identifies novel variants for coronary artery disease. PLoS Genet. 2011;7(9):e1002260.
  • Mann KG, Kalafatis M. Factor V. A combination of Dr Jekyll and Mr Hyde. Blood. 2003;10:20-30.
  • Camire RM, Bos MH. The molecular basis of factor V and VIII procofactor activation. J Thromb Haemost. 2009;7(12):1951-1.
  • Castoldi E, Brugge JM, Nicolaes GA, Girelli D, Tans G, Rosing J. Impaired APC cofactor activity of Factor V plays a major role in the APC resistance associated with the Factor V Leiden (R506Q) and R2 (H1299 R) mutations. Blood. 2004;103:4173-4179.
  • Tang L, Hu Y. Ethnic diversity in the genetics of venous thromboembolism. Thromb Haemost. 2015;114(5):901-909.
  • Var A, Utuk O, Akcali S, Sanlidag T, Uyanik BS, Dinc G. Impact of hemostatic gene single point mutations in patients with nondiabetic coronary artery disease. Mol Biol Rep. 2009;36(8):2235-2243.
  • Taymaz H, Erarslan S, Oner ET, Alkan T, Agirbasli M, Kirdar B. Sequence variations within the genes related to hemostatic imbalance and their impact on coronary artery disease in Turkish population. Thromb Res. 2007;119(1):55-62.
  • Thorelli E, Kaufman RJ, Dahlback B. Cleavage of Factor V at Arg 506 by activated protein C and the expression of anticoagulant activity of Factor V. Blood. 1999;93(8):2552-2558.
  • Boroumand M, Pourgholi L, Ziaee S, Anvari MS, Jalali A, Goodarzynejad H. The association between Factor V Leiden with the presence and severity of coronary artery disease. Clin Biochem. 2014;47(6):356-360.
  • Amara A, Mrad M, Sayeh A, et al. Association of FV G1691A Polymorphism but not A4070G with coronary artery disease. Clinical and Applied Thrombosis/Hemostasis. 2018;24(2):330-337.
  • Gardemann A, Arsic T, Katz N, Tillmanns H, Hehrlein FW, Haberbosch W. The factor II G20210A and factor V G1691A gene transitions and coronary heart disease. Thromb Haemost. 1999;81(2):208-213.
  • Irani Hakime N, Tamim H, Elias G, et al. Factor V R506Q mutation-Leiden: an independent risk factor for venous thrombosis but not coronary artery disease. J Thromb Thrombolysis. 2001;11(2):111-116.
  • Gemmati D, Occhionorelli S, Tisato V, et al. Inherited genetic predispositions in F13A1 and F13B genes predict abdominal adhesion formation: identification of gender prognostic indicators. Sci Rep. 2018;8:16916.
  • Franco RF, Reitsma PH, Lourenço D, et al. Factor XIII Val34Leu is a genetic factor involved in the etiology of venous thrombosis. Thromb. Haemost. 1999;81(5):676-9.
  • Ansani L, Marchesini J, Pestelli G, et al. F13A1 gene variant (V34L) and residual circulating FXIIIA levels predict short-and long-term mortality in acute myocardial infarction after coronary angioplasty. Int. J. Mol. Sci. 2018;19(9): 2766.
  • Balogh L, Katona É, Mezei ZA, et al. Effect of factor XIII levels and polymorphisms on the risk of myocardial infarction in young patients. Mol and Cell Biochem. 2018;448(1-2):199-209.
  • Mahley RW, Rall SC Jr. Apolipoprotein E: far more than a lipid transport protein. Annual review of genomics and human genetics. 2000;1:507–537.
  • Luo JQ, Ren H, Banh HL, et al.The associations between Apolipoprotein E Gene Epsilon2/Epsilon3/Epsilon4 polymorphisms and the risk of coronary artery disease in patients with type 2 diabetes mellitus. Front Physiol. 2017;8:1031.
  • Zheng YM, Sun R, Li XY. Relationship between ApoE gene polymorphism and type 2 diabetes mellitus with its cardiovascular complications in Chinese. Chin. J. Endocrinol. Metab. 1998;14,11–14.
  • AlBacha JA, Khoury M, Mouawad C, et al. High incidence of ACE/PAI-1 in Association to a Spectrum of Other Polymorphic Cardiovascular Genes Involving PBMCs proinflammatory cytokines in hypertensive hypercholesterolemic patients: reversibility with a combination of ACE inhibitor and statin. PLoS One. 2015;10(5):e0127266.
  • Dhar S, Ray S, Dutta A, Sengupta B, Chakrabarti S. Polymorphism of ACE gene as the genetic predisposition of coronary artery disease in Eastern India. Indian Heart J. 2012;64(6):576–581.
  • Xi B, Ruiter R, Chen J, Pan H, Wang Y, Mi J. The ACE insertion/deletion polymorphism and its association with metabolic syndrome. Metabolism. 2012;61(6):891–897.
  • Choudhury I, Jothimalar R, Patra AK. Angiotensin converting enzyme gene polymorphism and its association with hypertension in South Indian population. Indian J Clin Biochem. 2012;27(3):265–279.
  • Kandpal V, Sachdeva MP, Saraswathy KN. An assessment study of CVD related risk factors in a tribal population of India. BMC Public Health. 2016;16(1):434.
  • Weingarz L, Schindewolf M, Schwonberg J, et al. Thrombophilia and risk of VTE recurrence according to the age at the time of first VTE manifestation. Vasa. 2015;44(4):313-3.
  • Khare P, Raj V, Chandra S, Agarwal S. Quantitative and qualitative assessment of DNA extracted from saliva for its use in forensic identification. J.Forensic Dent. Sci. 2014;6:81–85.
  • Rigat B, Hubert C, Alhenc Gelas F, Corvol P, Soubrier F. An insertion deletion polymorphism in the angiotensin I-converting enzyme gene accounting for half the variance of serum enzyme levels. J Clin Invest. 1990;86:1343-1346.
  • World Health Organization. The 10 Leading Causes of death in the from:http://www.who.int/mediacentre/factsheets/fs310/en/ 2014. Yayınlanma tarihi 2014. Erişim tarihi 2016.
  • Barquera S, Pedroza TA, Medina C, et al. Global overview of the epidemiology of atherosclerotic cardiovascular disease. Arch. Med.Res. 2015;46(5):328-338.
  • Sakamuri SSVP, Higashi Y, Sukhanov S, et al. TRAF3IP2 mediates atherosclerotic plaque development and vulnerability in ApoE mice. Atherosclerosis. 2016;19(16):30206-30214.
  • Eitzman DT, Westrick RJ, Shen Y, et al. Homozygosity for Factor V Leiden leads to enhanced thrombosis and atherosclerosis in mice. Circulation. 2005;111:1822-1825.
  • Ozbek U, Tangün Y. Frequency of Factor V Leiden (Arg506Gln) in Turkey. Br J Haematol. 1997;97(2):504-505.
  • El-Fattah AAA, Sadik NAH, Sedrak H, Battah A, Nabil M. Association of genetic variants of hemostatic genes with myocardial infarction in Egyptian patients. Gene. 2018;641:212-219.
  • Vokó Z, Bereczky Z, Katona É, Ádány R, Muszbek L. Factor XIII Val34Leu variant protects against coronary artery disease. A meta-analysis.Thromb Haemost. 2007;97(3):458-463.
  • Wang G, Zou Z, Ji X, Ni Q, Ma Z. Factor XIII-A Val34Leu polymorphism might be associated with myocardial infarction risk: an updated meta-analysis. Int J Clin Exp Med 2014;7(12):5547-5552.
  • Hancer VS, Diz KR, Bilge AK, et al. The Association Between Factor XIII Val34Leu polymorphism and early myocardial infarction. Circ J. 2006;70(3):239-242.
  • Sabbagh AS, Otrock ZK, Mahfoud ZR, Zaatari GS, Mahfouz RA. Angiotensin-converting enzyme gene polymorphism and allele frequencies in the Lebanese population: prevalence and review of the literature. Mol Biol Rep. 2007;34(1):47-52.
  • Xu M, Zhao J, Zhang Y, et al. Apolipoprotein E gene variants and risk of coronary heart disease: a meta-analysis. Biomed Res Int. 2016;3912175.
  • Zhao QR, Lei YY, Li J, Jiang N, Shi JP. Association between apolipoprotein E polymorphisms and premature coronary artery disease: a meta-analysis. Clin Chem Lab Med. 2017;55(2):284-298.
  • Yilmaz AH, Isbir S, Kurnaz O, Gormus U, Isbir T. Associations of Lipoprotein Lipase S447X and Apolipoprotein E Genotypes with Low-density Lipoprotein Subfractions in Turkish patients with coronary artery disease. In vivo. 2009;23(1):155-161.
  • Arslan İnce FD, Atay A, Köseoğlu M, Yeşil M, Deveci E. Relationship between severity of coronary artery disease and apolipoprotein E gene polymorphism. Anadolu Kardiyol. Derg. 2010;10(3):202-208.
  • Fawzy MS, Toraih EA, Aly NM, Fakhr EA, Badran DI, Hussein MH. Atherosclerotic and thrombotic genetic and environmental determinants in Egyptian coronary artery disease patients: a pilot study. BMC Cardiovasc Disord. 2017;17(1):26.

Giresun İlinde FV, FXIII, ACE, ApoE Gen Varyantlarının Prevelansı ve Koroner Arter Hastalığı Profiline Etkilerinin Araştırılması

Yıl 2019, Sayı: 9, 864 - 880, 31.12.2019
https://doi.org/10.38079/igusabder.590895

Öz

Amaç: Ateroskleroz kaynaklı Koroner Arter Hastalığı (KAH), multifaktöriyel ve poligenik kompleks bir hastalıktır. Son yıllarda yapılan çalışmalarda KAH ile ilişkili bazı genetik varyantlar öne çıkmıştır. Bizim çalışmamızda da Türkiye’de Giresun ilinde yaşayanlarda KAH’na yatkınlık sağlayabilecek olan genetik profili çıkarmak için kan koagülasyonu ve lipid metabolizması ile ilgili olarak dört gen bölgesi analiz edildi. Bu gen bölgeleri FV-Leiden (rs6025) (FVL), FXIII 163G> T (V34L) (rs:5985), ACE (Angiotensin-converting enzyme) (rs1799752 I/D polimorfizm), ApoE (Apolipoprotein-E) (rs7412 ve rs429358) şeklindedir.

Yöntem: Çalışmamızda 89 kontrol ve 174 hasta kullanılmıştır. ApoE gerçek zamanlı polimeraz zincir reaksiyonu (RT-PCR), FVL, FXIII, ACE PCR yöntemi kullanılarak genotipler belirlenmiştir.

Bulgular: Hasta grubunda ACE D ve ApoE e4 allel frekansları kontrol grubunda düşük saptandı (sırasıyla p=0,026 ve p=0,015). Hasta grubunda, FV A allel taşıyanlar (GA+AA) GG genotipine sahip bireylerle karşılaştırıldığında serum total kolesterol (p=0,038) ve LDL-K (p=0,054) düzeyleri yüksek gözlendi. MI geçirmeyenlerde FXIII L allel varlığı % 31,9’dur. L allel varlığı MI riskine karşı koruyucu etki göstermektedir (p=0,06, OR=0,464, %95 CI=0,202-1,06).

Sonuç: Çalışmamızda araştırılan gen varyantlarının hiçbirinin direkt olarak KAH gelişimi ile ilişkisini gözlemlemedik. Ancak araştırılan gen varyantlarının dolaylı olarak KAH risk faktörlerine etkili olabilecekleri çalışmamızdan elde ettiğimiz çıkarımlardandır.

Kaynakça

  • Mozaffarian D, Benjamin EJ, Go AS, et al. American Heart Association statistics committee; stroke statistics subcommittee. Executive summary: heart disease and stroke statistics-2016 update: A report from the American Heart Association. Circulation. 2016;133(4):447-454.
  • Stranger BE, Forrest MS, Dunning M, et al. Relative impact of nucleotide and copy number variation on gene expression phenotypes. Science. 2007;315(5813):848-853.
  • Peden JF, Farrall, M. Thirty-five common variants for coronary artery disease: the fruits of much collaborative labour. Hum. Mol. Genet. 2011;20(R2):R198-205.
  • Wray NR, Goddard ME. Multi-locus models of genetic risk of disease. Genome Med. 2010;2(2):10
  • Consortium IKC. Large-scale gene-centric analysis identifies novel variants for coronary artery disease. PLoS Genet. 2011;7(9):e1002260.
  • Mann KG, Kalafatis M. Factor V. A combination of Dr Jekyll and Mr Hyde. Blood. 2003;10:20-30.
  • Camire RM, Bos MH. The molecular basis of factor V and VIII procofactor activation. J Thromb Haemost. 2009;7(12):1951-1.
  • Castoldi E, Brugge JM, Nicolaes GA, Girelli D, Tans G, Rosing J. Impaired APC cofactor activity of Factor V plays a major role in the APC resistance associated with the Factor V Leiden (R506Q) and R2 (H1299 R) mutations. Blood. 2004;103:4173-4179.
  • Tang L, Hu Y. Ethnic diversity in the genetics of venous thromboembolism. Thromb Haemost. 2015;114(5):901-909.
  • Var A, Utuk O, Akcali S, Sanlidag T, Uyanik BS, Dinc G. Impact of hemostatic gene single point mutations in patients with nondiabetic coronary artery disease. Mol Biol Rep. 2009;36(8):2235-2243.
  • Taymaz H, Erarslan S, Oner ET, Alkan T, Agirbasli M, Kirdar B. Sequence variations within the genes related to hemostatic imbalance and their impact on coronary artery disease in Turkish population. Thromb Res. 2007;119(1):55-62.
  • Thorelli E, Kaufman RJ, Dahlback B. Cleavage of Factor V at Arg 506 by activated protein C and the expression of anticoagulant activity of Factor V. Blood. 1999;93(8):2552-2558.
  • Boroumand M, Pourgholi L, Ziaee S, Anvari MS, Jalali A, Goodarzynejad H. The association between Factor V Leiden with the presence and severity of coronary artery disease. Clin Biochem. 2014;47(6):356-360.
  • Amara A, Mrad M, Sayeh A, et al. Association of FV G1691A Polymorphism but not A4070G with coronary artery disease. Clinical and Applied Thrombosis/Hemostasis. 2018;24(2):330-337.
  • Gardemann A, Arsic T, Katz N, Tillmanns H, Hehrlein FW, Haberbosch W. The factor II G20210A and factor V G1691A gene transitions and coronary heart disease. Thromb Haemost. 1999;81(2):208-213.
  • Irani Hakime N, Tamim H, Elias G, et al. Factor V R506Q mutation-Leiden: an independent risk factor for venous thrombosis but not coronary artery disease. J Thromb Thrombolysis. 2001;11(2):111-116.
  • Gemmati D, Occhionorelli S, Tisato V, et al. Inherited genetic predispositions in F13A1 and F13B genes predict abdominal adhesion formation: identification of gender prognostic indicators. Sci Rep. 2018;8:16916.
  • Franco RF, Reitsma PH, Lourenço D, et al. Factor XIII Val34Leu is a genetic factor involved in the etiology of venous thrombosis. Thromb. Haemost. 1999;81(5):676-9.
  • Ansani L, Marchesini J, Pestelli G, et al. F13A1 gene variant (V34L) and residual circulating FXIIIA levels predict short-and long-term mortality in acute myocardial infarction after coronary angioplasty. Int. J. Mol. Sci. 2018;19(9): 2766.
  • Balogh L, Katona É, Mezei ZA, et al. Effect of factor XIII levels and polymorphisms on the risk of myocardial infarction in young patients. Mol and Cell Biochem. 2018;448(1-2):199-209.
  • Mahley RW, Rall SC Jr. Apolipoprotein E: far more than a lipid transport protein. Annual review of genomics and human genetics. 2000;1:507–537.
  • Luo JQ, Ren H, Banh HL, et al.The associations between Apolipoprotein E Gene Epsilon2/Epsilon3/Epsilon4 polymorphisms and the risk of coronary artery disease in patients with type 2 diabetes mellitus. Front Physiol. 2017;8:1031.
  • Zheng YM, Sun R, Li XY. Relationship between ApoE gene polymorphism and type 2 diabetes mellitus with its cardiovascular complications in Chinese. Chin. J. Endocrinol. Metab. 1998;14,11–14.
  • AlBacha JA, Khoury M, Mouawad C, et al. High incidence of ACE/PAI-1 in Association to a Spectrum of Other Polymorphic Cardiovascular Genes Involving PBMCs proinflammatory cytokines in hypertensive hypercholesterolemic patients: reversibility with a combination of ACE inhibitor and statin. PLoS One. 2015;10(5):e0127266.
  • Dhar S, Ray S, Dutta A, Sengupta B, Chakrabarti S. Polymorphism of ACE gene as the genetic predisposition of coronary artery disease in Eastern India. Indian Heart J. 2012;64(6):576–581.
  • Xi B, Ruiter R, Chen J, Pan H, Wang Y, Mi J. The ACE insertion/deletion polymorphism and its association with metabolic syndrome. Metabolism. 2012;61(6):891–897.
  • Choudhury I, Jothimalar R, Patra AK. Angiotensin converting enzyme gene polymorphism and its association with hypertension in South Indian population. Indian J Clin Biochem. 2012;27(3):265–279.
  • Kandpal V, Sachdeva MP, Saraswathy KN. An assessment study of CVD related risk factors in a tribal population of India. BMC Public Health. 2016;16(1):434.
  • Weingarz L, Schindewolf M, Schwonberg J, et al. Thrombophilia and risk of VTE recurrence according to the age at the time of first VTE manifestation. Vasa. 2015;44(4):313-3.
  • Khare P, Raj V, Chandra S, Agarwal S. Quantitative and qualitative assessment of DNA extracted from saliva for its use in forensic identification. J.Forensic Dent. Sci. 2014;6:81–85.
  • Rigat B, Hubert C, Alhenc Gelas F, Corvol P, Soubrier F. An insertion deletion polymorphism in the angiotensin I-converting enzyme gene accounting for half the variance of serum enzyme levels. J Clin Invest. 1990;86:1343-1346.
  • World Health Organization. The 10 Leading Causes of death in the from:http://www.who.int/mediacentre/factsheets/fs310/en/ 2014. Yayınlanma tarihi 2014. Erişim tarihi 2016.
  • Barquera S, Pedroza TA, Medina C, et al. Global overview of the epidemiology of atherosclerotic cardiovascular disease. Arch. Med.Res. 2015;46(5):328-338.
  • Sakamuri SSVP, Higashi Y, Sukhanov S, et al. TRAF3IP2 mediates atherosclerotic plaque development and vulnerability in ApoE mice. Atherosclerosis. 2016;19(16):30206-30214.
  • Eitzman DT, Westrick RJ, Shen Y, et al. Homozygosity for Factor V Leiden leads to enhanced thrombosis and atherosclerosis in mice. Circulation. 2005;111:1822-1825.
  • Ozbek U, Tangün Y. Frequency of Factor V Leiden (Arg506Gln) in Turkey. Br J Haematol. 1997;97(2):504-505.
  • El-Fattah AAA, Sadik NAH, Sedrak H, Battah A, Nabil M. Association of genetic variants of hemostatic genes with myocardial infarction in Egyptian patients. Gene. 2018;641:212-219.
  • Vokó Z, Bereczky Z, Katona É, Ádány R, Muszbek L. Factor XIII Val34Leu variant protects against coronary artery disease. A meta-analysis.Thromb Haemost. 2007;97(3):458-463.
  • Wang G, Zou Z, Ji X, Ni Q, Ma Z. Factor XIII-A Val34Leu polymorphism might be associated with myocardial infarction risk: an updated meta-analysis. Int J Clin Exp Med 2014;7(12):5547-5552.
  • Hancer VS, Diz KR, Bilge AK, et al. The Association Between Factor XIII Val34Leu polymorphism and early myocardial infarction. Circ J. 2006;70(3):239-242.
  • Sabbagh AS, Otrock ZK, Mahfoud ZR, Zaatari GS, Mahfouz RA. Angiotensin-converting enzyme gene polymorphism and allele frequencies in the Lebanese population: prevalence and review of the literature. Mol Biol Rep. 2007;34(1):47-52.
  • Xu M, Zhao J, Zhang Y, et al. Apolipoprotein E gene variants and risk of coronary heart disease: a meta-analysis. Biomed Res Int. 2016;3912175.
  • Zhao QR, Lei YY, Li J, Jiang N, Shi JP. Association between apolipoprotein E polymorphisms and premature coronary artery disease: a meta-analysis. Clin Chem Lab Med. 2017;55(2):284-298.
  • Yilmaz AH, Isbir S, Kurnaz O, Gormus U, Isbir T. Associations of Lipoprotein Lipase S447X and Apolipoprotein E Genotypes with Low-density Lipoprotein Subfractions in Turkish patients with coronary artery disease. In vivo. 2009;23(1):155-161.
  • Arslan İnce FD, Atay A, Köseoğlu M, Yeşil M, Deveci E. Relationship between severity of coronary artery disease and apolipoprotein E gene polymorphism. Anadolu Kardiyol. Derg. 2010;10(3):202-208.
  • Fawzy MS, Toraih EA, Aly NM, Fakhr EA, Badran DI, Hussein MH. Atherosclerotic and thrombotic genetic and environmental determinants in Egyptian coronary artery disease patients: a pilot study. BMC Cardiovasc Disord. 2017;17(1):26.
Toplam 46 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Klinik Tıp Bilimleri
Bölüm Makaleler
Yazarlar

Ayşegül Başak Akadam-teker 0000-0003-3618-0560

Erhan Teker Bu kişi benim 0000-0002-0234-7548

Hülya Yılmaz Aydoğan 0000-0002-8837-6664

Aynur Dağlar Aday 0000-0001-8072-0646

Yayımlanma Tarihi 31 Aralık 2019
Kabul Tarihi 16 Ekim 2019
Yayımlandığı Sayı Yıl 2019 Sayı: 9

Kaynak Göster

JAMA Akadam-teker AB, Teker E, Yılmaz Aydoğan H, Dağlar Aday A. Giresun İlinde FV, FXIII, ACE, ApoE Gen Varyantlarının Prevelansı ve Koroner Arter Hastalığı Profiline Etkilerinin Araştırılması. IGUSABDER. 2019;:864–880.

 Alıntı-Gayriticari-Türetilemez 4.0 Uluslararası (CC BY-NC-ND 4.0)