Özgür ÇELEBİ, Demet ÇELEBİ, Ali TAGHİZADEHGHALEHJOUGHİ, Sümeyye BAŞER, Mustafa Can GÜLER, Serkan YILDIRIM

Klebsiella pneumoniae Enfeksiyon Modelinde HepG2 Hücre Hattında Bor Bileşiklerinin Antibakteriyel Etkisi ve Biyofilm Oluşumuna Etkilerinin Değerlendirilmesi

Amaç: Klebsiella pneumoniae bağışıklığı baskılanmış hastalarda hastane kaynaklı pnömoniye, idrar yolu enfeksiyonuna ve bakteriyemiye neden olur. Son zamanlarda daha sık görülen Klebsiella pneumoniae, insanlarda piyojenik karaciğer apselerine ve hematojen metastatik yayılımın yanı sıra antibiyotik direncine de neden olmaktadır. Gelişen antibiyotik direnci karaciğer enfeksiyonlarının tedavisini zorlaştırmaktadır. Bizde bu çalışmamızda Klebsiella pneumoniae 700603 tarafından oluşturulan bir enfeksiyon modelinde bor bileşiklerinin etkisini değerlendirmeyi amaçladık. Gereç ve Yöntem: Minimum inhibitör konsantrasyonu ve fraksiyonel inhibitör konsantrasyon çalışmaları, direnç gen seviyeleri ve HepG2 hücre analizleri yapılarak değerlendirildi. Bulgular: Bor bileşenleri sodyum perborat monohidrat ve etidot için sırasıyla düşük ve yüksek minimal inhibitör konsantrasyonu değerleri tespit edildi. Ayrıca sodyum perborat monohidrat biyofilm oluşumu ve direnç genleri üzerinde de etkili olduğu belirlendi. Bulgularımız, bor bileşiklerinin kombinasyon halinde kullanıldığında daha etkili olduğunu göstermiştir. Hücresel çalışmada oluşturulan toksisite modelinde, bor bileşiklerinin sitotoksik etkisi antibakteriyel etkilerinden dolayı azalmıştır. Sonuç: Bor bileşiklerinin etkili olduğu ve birlikte kullanıldıklarında olumlu etkinin arttığı görülmektedir.

Anahtar Kelimeler:

Bor bileşenleri, HEPG2, Klebsiella pneumoniae, Minimum inhibitör konsantrasyon

The Antibacterial Effect of Boron Compounds and Evaluation of the Effects on Biofilm Formation in the Infection Model of Klebsiella pneumoniae on the HepG2 Cell Line

Aim: Klebsiella pneumoniae causes hospital-acquired pneumonia, urinary tract infection and bacteremia in immunocompromised patients. Klebsiella pneumoniae, which has become more common recently, causes antibiotic resistance as well as pyogenic liver abscesses and hematogenous metastatic spread in humans. Developing antibiotic resistance complicates the treatment of liver infections. In our study, we aimed to evaluate the effect of boron compounds in an infection model created by Klebsiella pneumoniae 700603. Materials and Methods: Minimum inhibitory concentration and fractional inhibitory concentration studies, resistance gene levels, and HepG2 cell analyses were performed and evaluated. Results: We determined the low and high minimum inhibitory concentration values of boron components, sodium perborate monohydrate and etidote, respectively. In addition, sodium perborate monohydrate is also effective on biofilm formation and resistance genes. Our findings have shown that boron compounds are more effective when used in a combination. In the toxicity model created in the cellular study, the boron compounds cytotoxic effect decreased due to their antibacterial effects. Conclusion: İt seems that boron compounds are effective, and the positive effect increases when used together.

Keywords:

HEPG2, Klebsiella pneumoniae, Minimum inhibitory concentration, Boron components,

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1- Siu LK, Yeh KM, Lin JC, Fung CP, Chang FY. Klebsiella pneumoniae liver abscess: a new invasive syndrome.The Lancet.Infectious diseases,2012;2(11):881–7.
2- Podschun R, Ullmann U.Klebsiella spp as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev. 1998; 11: 589-603.
3- Fang CT, Chuang YP, Shun CT, Chang SC, Wang JT. A novel virulence gene in Klebsiella pneumoniae strains causing primary liver abscess and septic metastatic complications. J Exp Med. 2004;199:697-705
4- Thom BT. Klebsiella in faeces Lancet, 1970;2:1033.
5- Yu WL, Ko WC, Cheng KC, Lee CC, Lai CC, Chuang YC. Comparison of prevalence of virulence factors for Klebsiella pneumoniae liver abscesses between isolates with capsular K1/K2 and non-K1/K2 serotypes Diagn Microbiol Infect Dis. 2008; 62: 1-6.
6- Yeh KM, Chang FY, Fung CP, Lin JC, Siu LK. magA is not a specific virulence gene for Klebsiella pneumoniae strains causing liver abscess but is part of the capsular polysaccharide gene cluster of K pneumoniae serotype K. J Med Microbiol. 2006;55: 803-4.
7- Yu WL, Ko WC, Cheng KC. et al. Association between rmpA and magA genes and clinical syndromes caused by Klebsiella pneumoniae in Taiwan. Clin Infect Dis. 2006;42:1351-8.
8- Nassif X, Fournier JM, Arondel J, Sansonetti PJ. Mucoid phenotype of Klebsiella pneumoniae is a plasmid-encoded virulence factor.Infect Immun, 1989; 57:546-52.
9- Nassif X, Sansonetti PJ. Correlation of the virulence of Klebsiella pneumoniae K1 and K2 with the presence of a plasmid encoding aerobactin. Infect Immun.1986; 54: 603-8
10- Tacconelli E, Carrara E, Savoldi A, Harbarth S, Mendelson M, Monnet DL, et al. Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis. 2018;18:318–27.
11- Wencewicz TA. New antibiotics from Nature’s chemical inventory. Bioorg Med Chem. 2016;24:6227–52.
12- Butler MS, Paterson DL. Antibiotics in the clinical pipeline in October 2019. J Antibiot (Tokyo) 2020;73:329–364.
13- Crompton IE, Cuthbert BK, Lowe G, et al. β-Lactamase inhibitors. The inhibition of serine β-lactamases. Biochem J. 1988; 251:453-9.
14- Kiener PA, Waley SG. Reversible inhibitors of penicillinases, Biochem J, 1978; 169:197-204.
15- Chen Y, Shoichet B, Bonnet R. Structure, function, and inhibition along the reaction coordinate of CTX-M β-lactamases. J Am Chem Soc. 2005; 127:5423-34.
16- Demirci S, Dogan A, Karakus E, Halici Z, Topcu A, Demirci E, et al. Boron and Poloxamer (F68 and F127) Containing Hydrogel Formulation for Burn Wound Healing. Biol Trace Elem Res. 2015;168(1):169-80.
17- Nzietchueng RM, Dousset B, Franck P, Benderdour M, Nabet P, Hess K. Mechanisms implicated in the effects of boron on wound healing. J Trace Elem Med Biol. 2002;16(4):239-44.
18- Coudron PE. Inhibitor-based methods for detection of plasmid-mediated AmpC β-lactamases in Klebsiella spp., Esche-richia coli, and Proteus mirabilis.J Clin Microbiol.2005; 43:4163-7.
19- Doi Y, Paterson DL. Detection of plasmid-mediated class C β-lactamases, Int J Infect Dis, 2007; 11:191-7.
20- Livermore DM, Warner M, Mushtaq S. Evaluation of the chromogenic Cica-β-Test for detecting extended-spectrum, AmpC and metallo-β-lactamases. J Antimicrob Chemother. 2007; 60:1375-9.
21- Pasteran FG, Otaegui L, Guerriero L, et al. Klebsiella pneumoniae carbapenemase-2, Buenos Aires, Argentina, Emerg In-fect Dis, 2008; 14:1178-80.
22- Tsakris A, Kristo I, Poulou A. et al. First occurrence of KPC-2-possessing Klebsiella pneumoniae in a Greek hospital and recommendation for detection with boronic acid disc tests. J Antimicrob Chemother.2008;62:1257-60.
23- Tsakris A, Kristo I, Poulou A. et al. Evaluation of boronic acid disc tests for differentiating KPC-possessing Klebsiella pneumoniae isolates in the clinical laboratory, J Clin Microbiol, 2009; 47: 362-7.
24- Eidam O, Romagnoli C, Dalmasso G, Barelier S, Caselli E, Bonnet R, Shoichet BK, Prati F. Fragment-Guided Design of Subnanomolar β-Lactamase Inhibitors Active in Vivo. Proc. Natl. Acad. Sci. USA. 2012;109:17448–53.
25- Argın S, Gülerı̇m M, Şahı̇n F. Development of antimicrobial gelatin films with boron derivatives. Turk J Biol.2019; 43: 47-57.