Investigation of the Protective Effects of Graviola (Annona muricata), Moringa (Moringa oleifera Lam.) and Lactobacillus gasseri in a Pseudomonas aeruginosa Fibroblast Wound Model

Öz

Objective: This study aimed to investigate the antimicrobial, anti-inflammatory, antioxidant, and anti-apoptotic effects of Graviola (Annona muricata), Moringa oleifera, and Lactobacillus gasseri, alone and in combination, against Pseudomonas aeruginosa in vitro wound infection model. Methods: Antibacterial activity tests against Pseudomonas aeruginosa was determined using the Kirby-Bauer disk diffusion method, MIC/MBC, and FIC methods. Biofilm inhibition was measured by crystal violet staining. A scratch wound model using fibroblast cell line was infected with Pseudomonas aeruginosa. Following treatment with the extracts, cell viability, oxidative stress, cytokine levels (IL-1beta, IL-10 via ELISA), and apoptotic gene expression (BAX, BCL-2 via qPCR) were evaluated. Results: The triple combination (Graviola + Moringa + Lactobacillus gasseri) showed the strongest antibacterial activity (23 mm zone; FIC = 0.25) and notable antibiofilm effects. In infected fibroblasts, cell viability increased to with combination treatment, approaching control levels. TOS levels decreased significantly while TAC increased, indicating reduced oxidative stress. IL-1beta levels were significantly lowered, and IL-10 levels were restored. Additionally, BAX gene expression decreased by 47%, and BCL-2 increased by 50% in the triple treatment group, suggesting protection against apoptosis. Conclusion: The combined application of Graviola, Moringa oleifera, and Lactobacillus gasseri exerts synergistic antimicrobial, anti-inflammatory, antioxidant, and anti-apoptotic effects against Pseudomonas aeruginosa induced wound infection. These results will shed light on the search for alternative natural therapeutic strategies, particularly in the treatment of chronic wounds associated with antibiotic-resistant pathogens.

Anahtar Kelimeler

• Antibacterial effect
• Apoptosis
• Biofilm inhibition
• Oxidative stress
• Wound model

Graviola (Annona muricata), Moringa (Moringa oleifera Lam.) ve Lactobacillus gasseri'nin Pseudomonas aeruginosa Fibroblast Yara Modelinde Koruyucu Etkilerinin İncelenmesi

Öz

Amaç: Bu çalışmada, Graviola (Annona muricata), Moringa oleifera ve Lactobacillus gasseri'nin tekli ve kombinasyon halinde, Pseudomonas aeruginosa'ya karşı antimikrobiyal, antiinflamatuar, antioksidan ve antiapoptotik etkileri in vitro yara enfeksiyon modelinde araştırılmıştır. Yöntemler: Pseudomonas aeruginosa'ya karşı antibakteriyel aktivite testleri; Kirby-Bauer disk difüzyon yöntemi, MIC/MBC ve FIC yöntemleri kullanılarak belirlenmiştir. Biyofilm inhibisyonu kristal violet boyama yöntemiyle değerlendirilmiştir. Fibroblast hücre hattı kullanılarak oluşturulan çizik yara modeli Pseudomonas aeruginosa ile enfekte edilmiştir. Tedavi sonrası hücre canlılığı, oksidatif stres, sitokin düzeyleri (IL-1beta, IL-10, ELISA ile) ve apoptotik gen ekspresyonları (BAX, BCL-2, qPCR ile) analiz edilmiştir. Sonuçlar: Graviola + Moringa + Lactobacillus gasseri üçlü kombinasyonu en güçlü antibakteriyel aktiviteyi (23 mm zon; FIC = 0.25) ve anlamlı biyofilm inhibisyonunu göstermiştir. Enfekte fibroblastlarda, kombinasyon tedavisi ile hücre canlılığı artmış ve kontrol düzeylerine yaklaşmıştır. TOS seviyeleri anlamlı şekilde düşerken, TAC artmıştır; bu durum azalan oksidatif stresi göstermektedir. IL-1beta düzeyleri önemli ölçüde azalmış, IL-10 seviyeleri ise normale dönmüştür. Ayrıca, BAX gen ekspresyonu %47 azalmış, BCL-2 ise %50 artmıştır; bu durum apoptozisin önlendiğini göstermektedir. Tartışma: Graviola, Moringa oleifera ve Lactobacillus gasseri'nin kombinasyonu, Pseudomonas aeruginosa indüklenmiş yara enfeksiyonuna karşı sinerjik antimikrobiyal, antiinflamatuar, antioksidan ve antiapoptotik etkilere sahiptir. Bu bulgular, özellikle antibiyotik dirençli patojenlerle ilişkili kronik yaraların tedavisinde alternatif doğal bir terapötik strateji arayışına ışık tutacaktır.

Anahtar Kelimeler

• Antibakteriyel etki
• Apoptozis
• Biyofilm inhibisyonu
• Oksidatif stres
• Yara Modeli.

Kaynakça

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