Mikrobiyal kondroitin sülfatın farklı Candida suşları üzerindekiantifungal etkinliğinin deneysel vulvovajinal kandidiyazis ratmodelinde değerlendirilmesi
Küçük Resim Yok
Tarih
2024
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Kondroitin Sülfat (KS), vücutta yapısal ve sinyal iletim fonksiyonlarına sahip sülfatlanmış bir glikozaminoglikandır. Ayrıca nutrasötik ve farmasötik bir ajan olarak dünya genelinde yaygın şekilde kullanılmaktadır. KS'nin kemik mineralizasyonu, sinir gelişimi ve anjiyogeneze katkı sağladığı, antioksidan ve antienflamatuvar özellikler gösterdiği kanıtlanmıştır. Candida türleri, yüzeysel ve mukozal enfeksiyonlardan, invaziv kandidemi ve metastatik organ tutulumuna kadar geniş bir klinik yelpazede enfeksiyonlara yol açarak, insanlarda morbidite ve mortalitenin başlıca nedenleri arasında yer alır. Vulvovajinal kandidiyazis, kadınların % 70-75'ini hayatları boyunca en az bir kez etkileyen sık rastlanan bir vajinal enfeksiyon türüdür. Candida enfeksiyonlarına karşı geliştirilen antifungal tedavi seçenekleri konusunda halen önemli bilgi eksiklikleri bulunmaktadır. Bu tez çalışmasında, biyoteknolojik yöntemler kullanılarak rekombinant E. coli (C2987) suşundan üretilen mikrobiyal KS'nin (MKS), deneysel vulvovajinal kandidiyazis rat modelinde çeşitli Candida suşlarına karşı antifungal etkileri araştırılmış ve diğer yaygın antifungal ajanlarla karşılaştırılmıştır. Kondroitin sentezinde rol oynayan kfoA, kfoC, kfoF genleri ve Vitreoscilla hemoglobin genini (vgb) içeren pETM6-PACF-vgb plazmidinin E. coli suşuna transforme edilmesiyle üretilen MKS, NMR ve HPLC analizleri ile yapısal bütünlüğü ve saflık açısından doğrulanmıştır. Ayrıca, farklı oranlarda MKS içeren krem formülasyonlarının kimyasal yapısı FTIR analizleri ile desteklenmiştir. Deneysel vulvovajinal kandidiyazis modelinde tedavi edilen gruplarda, özellikle % 0.5 ve %1 MKS kullanılan gruplarda, vajinal lavaj sıvısındaki Candida koloni sayıları ve sitokin düzeyleri, kontrol grubuna ve nistatin ile fentikonazol gibi antifungal ajanlarla tedavi edilen gruplara kıyasla istatistiksel olarak anlamlı derecede düşük bulunmuştur. MKS'nin düşük dozlarda dahi etkinlik göstermesi, bu ajanın vajinal enfeksiyonlara karşı potansiyel koruyucu ve tedavi edici etkisini desteklemektedir. Bu sonuçlar, MKS'nin klinik kullanımı ve daha fazla araştırılması gerektiğini vurgulamaktadır. Anahtar Kelimeler: Antifungal aktivite, Candida spp., Mikrobiyal Kondroitin Sülfat, Vajinal kandidiyazis
Chondroitin sulfate (CS) is a sulphated glycosaminoglycan with structural and signaling functions in the body. It is also widely used as a dietary supplement and active pharmaceutical agent worldwide. CS has been shown to contribute to bone mineralization, nerve development and angiogenesis, as well as to exhibit antioxidant and anti-inflammatory properties. Candida species cause a wide range of infections, from superficial and mucosal infections to invasive candidemia and metastatic organ involvement and are a major cause of morbidity and mortality in humans. Vulvovaginal candidiasis is a common vaginal infection affecting 70-75% of women at least once in their lifetime. There are still significant knowledge gaps regarding alternative antifungal treatment options for Candida infections. In this thesis, the antifungal activity of microbial CS (MCS) produced by biotechnological methods from the recombinant E. coli strain (C2987) was investigated in an experimental vulvovaginal candidiasis rat model against different Candida strains and compared with other commonly used antifungal agents. MCS was produced by transforming E. coli with the pETM6- PACF-vgb plasmid containing the kfoA, kfoC and kfoF genes involved in chondroitin synthesis as well as the Vitreoscilla hemoglobin gene (vgb). The structural integrity and purity of the product was confirmed by NMR and HPLC analyzes. In addition, the chemical composition of the cream formulations containing different concentrations of MCS was verified by FT-IR analyzes. In the experimental model of vulvovaginal candidiasis model, treatment groups, especially those treated with 0.5% and 1% MCS, showed statistically significant reductions in vaginal lavage fluid Candida colony counts and cytokine levels compared to the infected control group and those treated with known antifungal agents such as nystatin and fenticonazole. The efficacy of MCS, even at low doses, supports its potential protective and therapeutic effect against vaginal infections. These results emphasize the need for further investigation of MCS in clinical practice and its potential for wider use. Keywords: Antifungal activity, Candida spp., Microbial Chondroitin Sulfate, Vaginal candidiasis
Chondroitin sulfate (CS) is a sulphated glycosaminoglycan with structural and signaling functions in the body. It is also widely used as a dietary supplement and active pharmaceutical agent worldwide. CS has been shown to contribute to bone mineralization, nerve development and angiogenesis, as well as to exhibit antioxidant and anti-inflammatory properties. Candida species cause a wide range of infections, from superficial and mucosal infections to invasive candidemia and metastatic organ involvement and are a major cause of morbidity and mortality in humans. Vulvovaginal candidiasis is a common vaginal infection affecting 70-75% of women at least once in their lifetime. There are still significant knowledge gaps regarding alternative antifungal treatment options for Candida infections. In this thesis, the antifungal activity of microbial CS (MCS) produced by biotechnological methods from the recombinant E. coli strain (C2987) was investigated in an experimental vulvovaginal candidiasis rat model against different Candida strains and compared with other commonly used antifungal agents. MCS was produced by transforming E. coli with the pETM6- PACF-vgb plasmid containing the kfoA, kfoC and kfoF genes involved in chondroitin synthesis as well as the Vitreoscilla hemoglobin gene (vgb). The structural integrity and purity of the product was confirmed by NMR and HPLC analyzes. In addition, the chemical composition of the cream formulations containing different concentrations of MCS was verified by FT-IR analyzes. In the experimental model of vulvovaginal candidiasis model, treatment groups, especially those treated with 0.5% and 1% MCS, showed statistically significant reductions in vaginal lavage fluid Candida colony counts and cytokine levels compared to the infected control group and those treated with known antifungal agents such as nystatin and fenticonazole. The efficacy of MCS, even at low doses, supports its potential protective and therapeutic effect against vaginal infections. These results emphasize the need for further investigation of MCS in clinical practice and its potential for wider use. Keywords: Antifungal activity, Candida spp., Microbial Chondroitin Sulfate, Vaginal candidiasis
Açıklama
Fen Bilimleri Enstitüsü, Biyoloji Ana Bilim Dalı
Anahtar Kelimeler
Biyoloji, Biology
