In Vitro Effects of Hypericum pseudolaeve N. Robson Aqueous Extract on the Expression of Wound Healing-Related Genes in L929 Fibroblast Cells

Abstract

Wound healing is a complex process involving extracellular matrix remodeling, angiogenesis, and tissue regeneration. Although medicinal plants are widely recognized for their therapeutic potential, the molecular basis of their wound-healing activity remains incompletely understood. This study investigated the effects of the water-macerated extract of Hypericum pseudolaeve N. Robson (HWM) on the expression of wound healing–related genes in L929 fibroblast cells. Cells were treated with HWM extract (62 µg/mL) for 24 h and the expression of collagen, FGF7, TGF-β1, and VEGF was evaluated using qRT-PCR and ELISA methods. HWM treatment increased the expression of collagen, FGF7, TGF-β1, and VEGF by 2.0, 3.8, 47.9, and 3.6-fold, respectively, compared with untreated controls. ELISA analyses supported the qRT-PCR findings by demonstrating elevated secreted protein levels of the investigated markers. Among the analyzed genes, TGF-β1 exhibited the most pronounced statistically significant response in both methods. Overall, the results indicate that Hypericum pseudolaeve aqueous extract can promote the expression of key mediators involved in extracellular matrix formation, growth factor signaling, and angiogenesis, which may play role its wound-healing potential.

Keywords

• Hypericum pseudolaeve
• wound healing
• L929 fibroblasts
• qRT-PCR
• ELISA

Supporting Institution

Gazi University Scientific Research Projects Coordination Unit (Project No: 05/2017-16), the Scientific and Technological Research Council of Türkiye (TÜBİTAK) for financial support through the 2211/C National PhD Scholarship Program

Ethical Statement

Ethics committee approval is not required for this study

Thanks

The authors gratefully acknowledge the financial support provided by Gazi University Scientific Research Projects Coordination Unit (Project No: 05/2017-16). Bahar Kaptaner İğci sincerely thanks the Scientific and Technological Research Council of Türkiye (TÜBİTAK) for financial support through the 2211/C National PhD Scholarship Program in Priority Areas and is currently supported by the TÜBİTAK 2218 National Postdoctoral Research Fellowship Program.

Hypericum pseudolaeve N. Robson Sulu Ekstresinin L929 Fibroblast Hücrelerinde Yara İyileşmesi ile İlgili Genlerin Ekspresyonu Üzerine In Vitro Etkileri

Abstract

Yara iyileşmesi; ekstraselüler matriksin yeniden düzenlenmesi, anjiyogenez ve doku rejenerasyonunu kapsayan karmaşık bir biyolojik süreçtir. Tıbbi bitkiler terapötik potansiyelleri nedeniyle yaygın olarak kullanılmaktadır. Ancak bu bitkilerin yara iyileşmesi üzerindeki etkilerinin moleküler mekanizmaları henüz tam olarak açıklanamamıştır. Bu çalışmada, Hypericum pseudolaeve N. Robson'un su maserasyonu yöntemiyle hazırlanan ekstraktının (HWM), L929 fibroblast hücrelerinde yara iyileşmesiyle ilişkili genlerin ifadesi üzerindeki etkileri araştırılmıştır. L929 fibroblast hücreleri, 62 µg/mL konsantrasyondaki HWM ekstraktı ile 24 saat boyunca inkübe edilmiştir. Kolajen, fibroblast büyüme faktörü 7 (FGF7), dönüştürücü büyüme faktörü-β1 (TGF-β1) ve vasküler endotelyal büyüme faktörü (VEGF) genlerinin ekspresyon düzeyleri qRT-PCR ve ELISA yöntemleri kullanılarak belirlenmiştir. HWM uygulaması sonucunda kolajen, FGF7, TGF-β1 ve VEGF genlerinin ifade düzeyleri kontrol grubuna göre sırasıyla 2,0; 3,8; 47,9 ve 3,6 kat artmıştır. ELISA analizleri, incelenen belirteçlerin protein düzeylerindeki artışı ortaya koymuş ve qRT-PCR sonuçlarını desteklemiştir. İncelenen genler arasında en yüksek artış TGF-β1 ifadesinde belirlenmiştir. Elde edilen bulgular, HWM ekstraktının ekstraselüler matriks oluşumu, büyüme faktörü sinyal iletimi ve anjiyogenez süreçlerinde görev alan temel aracı moleküllerin ekspresyonunu artırdığını göstermektedir. Sonuç olarak, Hypericum pseudolaeve'nin yara iyileşmesini destekleyebilecek önemli bir biyolojik potansiyele sahip olduğu değerlendirilmiştir.

Keywords

• Hypericum pseudolaeve
• yara iyileşmesi
• L929 fibroblastları
• qRT-PCR
• ELISA

Supporting Institution

Gazi Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi, Türkiye Bilimsel ve Teknolojik Araştırma Kurumu’na (TÜBİTAK), 2211/C Öncelikli Alanlarda Ulusal Doktora Burs Programı

Thanks

Yazarlar, Gazi Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından sağlanan mali desteğe (Proje No: 05/2017-16) teşekkür ederler. Bahar Kaptaner İğci, Türkiye Bilimsel ve Teknolojik Araştırma Kurumu’na (TÜBİTAK), 2211/C Öncelikli Alanlarda Ulusal Doktora Burs Programı kapsamındaki mali desteği için içtenlikle teşekkür eder; kendisi hâlihazırda TÜBİTAK 2218 Ulusal Doktora Sonrası Araştırma Burs Programı tarafından desteklenmektedir.

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