Ajuga reptans L. Etanol Özütünün HCT116 Kolon Kanseri Hücrelerinde NF-κB ve eNOS Aktivitesinin Inhibisyonu Yoluyla Apoptotik ve Antiproliferatif Etkileri

Öz

Amaç: Bu çalışmanın amacı, Ajuga reptans L. (Lamiaceae) etanol ekstraktının, HCT116 insan kolorektal karsinom hücrelerinde hücre proliferasyonu, apoptotik yanıt, Kaspaz-3, NF-κB ve eNOS proteinlerinin ifade düzeyleri üzerine etkilerini araştırmaktır. Materyal ve Metot: A. reptans bitkisi Soxhlet cihazı kullanılarak etanol ile ekstrakte edilmiştir. Elde edilen ekstrakt, HCT116 ve HUVEC (sağlıklı kontrol) hücrelerine artan konsantrasyonlarda (0–800 µg/mL) uygulanmıştır. Hücre canlılığı, WST-1 testi ile 24 ve 48 saatlik sürelerde değerlendirilmiştir. Morfolojik değişiklikler inverted mikroskopi ile incelenmiştir. Kaspaz-3, NF-κB ve eNOS protein düzeylerinin kantifikasyonu için ELISA analizi uygulanmıştır. Bulgular: A. reptans ekstraktı, HCT116 hücrelerinde konsantrasyona ve süreye bağımlı sitotoksik etki göstermiştir; IC₅₀ değerleri sırasıyla 24 saatte 206.9 µg/mL, 48 saatte 147.5 µg/mL olarak hesaplanmıştır. HUVEC hücrelerinde minimum düzeyde sitotoksisite gözlenmiştir. Mikroskobik incelemeler, HCT116 hücrelerinde apoptozu düşündüren morfolojik değişiklikleri doğrulamıştır. ELISA analizleri, Caspase-3 düzeylerinde artış ve NF-κB ile eNOS düzeylerinde azalma göstermiştir. Sonuç: Ajuga reptans’ın etanolik ekstraktı, kolorektal kanser hücrelerinde seçici proliferasyon baskılanması, apoptoz indüksiyonu ve NF-κB ile eNOS sinyallerinin modülasyonu yoluyla antikanser etki göstermiştir. Bu bulgular, ekstraktın kolorektal kanser tedavisinde bitki kaynaklı tamamlayıcı bir ajan olarak potansiyelini ortaya koymaktadır.

Anahtar Kelimeler

• A. reptans
• endotel nitrik oksit sentaz (eNOS)
• kaspaz-3
• kolon kanseri
• nuclear factor kappa B (NF-κB)

Apoptotic and Antiproliferative Effects of Ajuga reptans L. Ethanol Extract via Inhibition of NF-κB and eNOS Activity in HCT116 Colon Cancer Cells

Abstract

Objective: This study aimed to investigate the ethanol extract of Ajuga reptans L. (Lamiaceae) on cell proliferation, apoptotic response, expression levels of Caspase-3, NF-κB and eNOS proteins on HCT116 human colorectal carcinoma cells. Materials and Methods: A. reptans was extracted using a Soxhlet apparatus with ethanol, and the obtained extract was applied to HCT116 and HUVEC cells (as healthy controls) in increasing concentrations (0–800 µg/mL). Cell viability was analysed by the WST-1 assay at 24 h and 48 h. Morphological changes were observed via inverted microscopy. ELISA was performed to quantify Caspase-3, NF-κB, and eNOS protein levels. Results: A. reptans ethanol extract showed concentration and duration-dependent cytotoxic effects on HCT116 cells, with IC₅₀ values of 206.9 µg/mL (24 h) and 147.5 µg/mL (48 h). Minimal cytotoxicity was observed in HUVEC cells. Microscopy confirmed morphological signs of apoptosis in HCT116 cells. ELISA results demonstrated increased Caspase-3 and decreased NF-κB and eNOS levels, indicating induction of apoptosis and suppression of pro-survival pathways. Conclusions: The ethanol extract of Ajuga reptans selectively inhibited the proliferation of colorectal cancer cells by apoptotic induction and modulation of NF-κB and eNOS signaling, suggesting its potential as a plant-based adjunctive agent in colorectal cancer therapy.

Keywords

• Ajuga reptans
• caspase-3
• colon cancer
• eNOS
• NF-κB

Kaynakça

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