LAKTOBASİLLERDEN ELDE EDİLEN HÜCRESİZ SÜZÜNTÜLERİN İNSAN ADENOKARSİNOM (HT-29) HÜCRE KÜLTÜRÜNDE CAMPYLOBACTER JEJUNİ’NİN ÜREME VE VİRÜLANS ÖZELLİKLERİ ÜZERİNE ETKİLERİ

Abstract

Amaç: Laktobasiller en yaygın kullanılan probiyotiklerdendir. Çalışmamızda, Lactobacillus acidophilus (La), L. fermentum (Lf), L. plantarum (Lp) ve L. rhamnosus (Lr)’nin hücresiz süzüntülerinin (CFS) C. jejuni suşlarının (81116 ve RM1221) üremesi, adezyonu ve invazyonu üzerine etkilerini inceledik. Aynı zamanda, CFS’lerin, C. jejuni suşlarının ve CFS+C. jejuni kombinasyonlarının HT-29 hücre canlılığındaki etkilerini araştırdık. Gereç ve Yöntem: Üreme ile adezif ve invazif bakterilerin sayıları, sırasıyla, spektrofotometrik ve koloni sayma yöntemleri ile belirlenmiştir. Çalışmamızda, Metil tiazolil difeni-tetrazolium bromid (MTT) deneyini, hücrelerin (HT-29) canlılığını belirlemede kullandık. Bulgular: İki ve dört saatlik inkübasyonlarda, tüm CFS’ler RM1221 suşunun, La ve Lp CFS’leri ise, 81116 suşunun üremesini anlamlı düzeyde azaltmıştır (sırasıyla, p<0,0001, p<0,05). Tüm CFS’ler (La hariç) 24 saatlik inkübasyonda her iki suşun da üremesini baskılamıştır. C. jejuni 81116’nin adezyonu tüm CFS’lerin varlığında istatistiksel olarak anlamlı düzeyde baskılanmıştır (p<0,0001). C. jejuni RM1221’nin adezyonu La ve Lr süzüntüleri varlığında istatistiksel olarak anlamlı düzeyde baskılanmıştır (sırasıyla, p<0,05 ve p<0,005). Suşların invazyon özellikleri süzüntülerin varlığında etkilenmemiştir. HT-29 hücrelerinin canlılığı hem C. jejuni’nin ve hem de her bir CFS’nin varlığında farklı yönlerde etkilenmiştir. Sonuç: Sonuçlarımız CFS’lerin C. jejuni’nin üreme ve adezyonunu temas süresine bağlı olarak baskıladıklarını göstermektedir. HT-29 hücrelerinin canlılığı inkübasyon süresi ve incelenen suşa bağlı olarak etkilenmiştir.

Keywords

• C.jejuni
• laktobasiller
• üreme
• adezyon
• invazyon
• hücre canlılığı

THE INFLUENCES OF LACTOBACILLUS CELL-FREE SUPERNATANTS ON GROWTH AND VIRULENCE PROPERTIES OF CAMPYLOBACTER JEJUNI IN HUMAN ADENOCARCINOMA (HT-29) CELL CULTURE

Abstract

Objective: Lactobacilli are the most commonly used probiotics. We examined the influence of cell-free supernatants (CFSs) of Lactobacillus acidophilus (La), L. fermentum (Lf), L. plantarum (Lp) and L. rhamnosus (Lr) on growth, adhesion and invasion of C. jejuni 81116 and RM1221 in human adenocarcinoma colon cells (HT-29). We also analyzed the influences of CFSs, C. jejuni and their combinations on HT-29 cell viability. Materials and Methods: Growth and adhesive-invasive bacteria counts were determined using the spectrophotometric method and colony counting method, respectively. We used methyl thiazolyl diphenyl-tetrazolium bromide (MTT) assay for detection of HT-29 cell viability. Results: During two and four hours of incubation, the growth of RM1221 was significantly decreased (p<0.0001) with the effects of the tested CFSs, while the decrease in growth of the 81116 strain was only significant (p<0.05) in the presence of La and Lp. All CFSs except La reduced the growth of both C. jejuni isolates at 24 hours of incubation. The adhesion of C. jejuni 81116 was significantly (p<0.0001) reduced in the presence of all CFSs. La and Lr statistically significantly (p<0.05 and p<0.005, respectively) reduced the adhesion of C. jejuni RM1221. Invasion of C. jejuni strains was shown not to be affected in presence of all CFSs. C. jejuni and each CFSs were found to influence the HT-29 viability differently. Conclusion: Our results suggest that CFSs have suppressive effects on the growth and adhesive properties of C. jejuni in a time-dependent manner. The viability of HT-29 depends on incubation time and which strain is tested.

Keywords

• C. jejuni
• lactobacilli
• growth
• adhesion
• invasion
• cell viability

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