j. fac. pharm. ankara Journal of Faculty of Pharmacy of Ankara University 1015-3918 2564-6524 Ankara University 10.33483/jfpau.1336857 Pharmaceutical Toxicology Farmasotik Toksikoloji PRANGOS HEYNIAE H. DUMAN & M. F. WATSON EKSTRELERİNİN HEPG2 HÜCRELERİNDEKİ SİTOTOKSİK VE APOPTOTİK ETKİLERİNİN ARAŞTIRILMASI INVESTIGATION OF CYTOTOXIC AND APOPTOTIC EFFECTS OF PRANGOS HEYNIAE H. DUMAN & M. F. WATSON EXTRACTS ON HEPG2 CELLS https://orcid.org/0000-0002-3239-4855 Arzuk Ege Ege University, Faculty of Pharmacy, Department of Toxicology https://orcid.org/0000-0002-5729-0796 Albayrak Gökay İzmir Katip Çelebi Üniversitesi, Eczacılık Fakültesi, Farmasötik Botanik AD https://orcid.org/0000-0002-9791-4399 Ergüç Ali İzmir Katip Çelebi Üniversitesi, Eczacılık Fakültesi, Farmasötik Toksikoloji AD https://orcid.org/0009-0001-8475-8067 Atış Ecrin EGE UNIVERSITY https://orcid.org/0009-0001-2006-5480 Tan İclal EGE UNIVERSITY https://orcid.org/0000-0002-3624-4811 Baykan Şüra Egei Üniversitesi, Eczacılık Fakültesi, Farmasötik BotanikAD 01 20 2024 48 1 46 55 08 02 2023 10 05 2023 Copyright © 1971, Journal of Faculty of Pharmacy of Ankara University 1971 Journal of Faculty of Pharmacy of Ankara University

Amaç: Bu çalışmanın amacı; Prangos Heyniae H. Duman & M. F. Watson kök ekstrelerinin insan karaciğer kanseri hücrelerindeki antikanser potansiyellerini araştırmak ve ekstre ile indüklenen sitotoksisitede rol oynayan moleküler mekanizmaları değerlendirmektir. Gereç ve Yöntem: P. heyniae kök ekstrelerin HepG2 hücrelerinin canlılığına olan olası etkilerini araştırmak amacıyla hücreler kloroform, n-hekzan ya da metanol ekstreleri ile inkübe edildi. MTT testi ile IC50 değerlerinin belirlenmesi sonrası, selektivite indeksleri nedeniyle n-hekzan ve metanol ekstreleri ile çalışmaya devam edilmedi. Yapı-biyolojik aktivite ilişkisini kurabilmek amacıyla kloroform ekstresinin kimyasal karakterizasyonu HPLC analizi ile gerçekleştirildi. Kloroform ekstresinin mitokondriyal membran potansiyeli ve kaspaz-3 aktivasyonu üzerindeki etkileri ileri deneylerle araştırıldı. Ayrıca, sitotoksisitede rol oynayan hücre ölüm yolunun belirlenmesi amacıyla selektif inhibitörler varlığında hücre canlılığı ölçüldü. Sonuç ve Tartışma: Kloroform ekstresi, HepG2 hücrelerinde canlılığın güçlü ve selektif inhibisyonuna neden oldu. Benzer sitotoksik etki n-hekzan ya da metanol ekstreleri ile saptanmadı. Kloroform ekstresi tarafından indüklenen sitotoksisite pan-kaspaz apoptoz inhibitörü varlığında önlendi. Ayrıca, HepG2 hücrelerinin kloroform ekstresi ile inkübasyonu, mitokondriyal membran potansiyelinde hasara ve kaspaz-3 aktivasyonuna neden oldu. Kloroform ekstresinin ana bileşenleri olarak oxypeucedanin, isoimperatorin ve osthole tespit edildi. Bu sonuçlar, kloroform ekstresindeki kumarin ve furanokumarin türevlerinin antikanser etki mekanizmasında apoptozun rol oynayabileceğini göstermektedir.

Objective: This study aims to investigate the anticancer potential of Prangos Heyniae H. Duman & M. F. Watson root extracts against human hepatoma cells, and examine the molecular mechanisms potentially involved in extract-induced cytotoxicity.Material and Method: HepG2 cells were treated with chloroform, n-hexane, or methanol extracts from roots of P. heyniae to investigate the possible effects on cell viability. Following the determination of IC50 values by the MTT test, n-hexane, and methanol extracts were excluded because of their selectivity indices. The chemical characterization of chloroform extract was performed by HPLC to understand the chemical composition-bioactivity relationship. Alterations induced by chloroform extract on mitochondrial membrane potential and caspase-3 activation were further investigated. In addition, cell viability was measured in the presence of different selective inhibitors of pathways to define the type of cell death pathway contributing to cytotoxicity.Result and Discussion: Chloroform extract but not n-hexane or methanol extracts led to strong and selective inhibition of cell viability on HepG2 cells. In addition, cytotoxicity increased by chloroform extract was only restored in the presence of a pan-caspase apoptosis inhibitor. Also, treatment of HepG2 cells with chloroform extract impaired mitochondrial membrane potential and led to significant caspase-3 activation. Oxypeucedanin, isoimperatorin, and osthole were detected as the major components of the chloroform extract. These results represent that apoptosis may be involved in the anticancer effect of coumarin and furanocoumarin derivatives in chloroform extract.

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