Ursolik asidin RANKL ile indüklenen osteoklast farklılaşması üzerindeki in vitro etkileri

Year 2024, Volume: 49 Issue: 3, 712 - 720, 30.09.2024

Mehmet Berköz , Maryenur Çetiner

https://doi.org/10.17826/cumj.1466896

Abstract

Amaç: Ursolik asit, izoprenoid birimlerden oluşan küçük bir pentasiklik triterpen molekülüdür. Ursolik asidin romatoid artrit tedavisinde etkili olduğu birçok çalışmada gösterilmiş olmasına rağmen, kemik kaybının neden olduğu kemik hastalıkları üzerine çok sınırlı araştırma yapılmıştır. Bu çalışma, kemikleri güçlendirmek ve yaşlılıkta canlılığı artırmak için osteoklast fonksiyonunu inhibe eden bitkisel ilaçlar bulmak amacıyla ursolik asidin osteoklast oluşumu üzerindeki etkilerini değerlendirmeyi amaçlamıştır.
Gereç ve Yöntem: Çalışmamızda RAW264.7 fare makrofajları kullanılmış ve hücreler osteoklastik farklılaşma için 100 ng/mL RANKL ile muamele edilmiştir. Daha sonra, ursolik asit tedavisinin hücre canlılığı, tartarat dirençli asit fosfataz (TRAP) oluşumu ve osteoklastik gen ekspresyon seviyeleri üzerindeki etkileri ölçülmüştür.
Bulgular: Sonuçlarımız ursolik asidin 2.5-10 µg/mL konsantrasyonlarında önemli bir sitotoksisite sergilemediğini göstermiştir (%3,2-9,8). Ayrıca, ursolik asit osteoklastların farklılaşmasını inhibe etmiş (%15,2-39,1) ve katepsin K (%3,8-22,3), TRAP (%16,3-48,7), matriks metalloproteinaz-9 (MMP-9) (%10,7-40,2), aktive T hücre sitoplazmik nükleer faktörü-1 (NFATc1) (%1,2-29,7), c-Fos (%0,9-13,8) ve mikroftalmi ile ilişkili transkripsiyon faktörü (MITF) (%2,2-21,6) gibi osteoklastik genlerin ekspresyonunu baskılamıştır.
Sonuç: Ursolik asidin RANKL ile indüklenen osteoklast farklılaşmasını inhibe ettiği gösterilmiştir ve bu nedenle ursolik asidin osteoporozun tedavisi ve önlenmesi için kullanılabileceği kanaatini taşımaktayız.

Keywords

Ursolik asit, RAW264.7 makrofajları, osteoporoz, osteoklast, RANKL

Project Number

TLO-2023-10609

In vitro effects of ursolic acid on RANKL-induced osteoclast differentiation

Abstract

Purpose: Ursolic acid is a small pentacyclic triterpene molecule composed of isoprenoid units. Although ursolic acid has been shown to be effective in the treatment of rheumatoid arthritis in many studies, very little research has been conducted on bone diseases caused by bone loss. The present study aimed to evaluate the effects of ursolic acid on osteoclast formation with the aim of finding herbal medicines that inhibit osteoclast function to strengthen bones and promote vitality in old age.
Materials and Methods: RAW264.7 murine macrophages were used in our study and cells were treated with 100 ng/mL RANKL for osteoclastic differentiation. The effects of ursolic acid treatment on cell viability, tartrate-resistant acid phosphatase (TRAP) formation and osteoclastic gene expression levels were then measured.
Results: Our results showed that ursolic acid did not exhibit significant cytotoxicity (3.2-9.8%) at concentrations of 2.5-10 µg/mL. Furthermore, ursolic acid inhibited osteoclast differentiation (15.2-39.1%) and suppressed the expression of osteoclastic genes such as cathepsin K (3.8-22.3%), TRAP (16.3-48. 7%), matrix metalloproteinase-9 (MMP-9) (10.7-40.2%), nuclear factor of activated T-cell cytoplasmic 1 (NFATc1) (1.2-29.7%), c-Fos (0.9-13.8%) and microphthalmia-associated transcription factor (MITF) (2.2-21.6%).
Conclusion: Ursolic acid has been shown to inhibit RANKL-induced osteoclast differentiation and therefore we believe that ursolic acid may be used for the treatment and prevention of osteoporosis.

Keywords

Ursolic acid, RAW264.7 macrophages, osteoporosis, osteoclast, RANKL

Ethical Statement

As this research is a cell culture study, ethics committee approval is not required. No conflict of interest was declared by the authors.

Supporting Institution

This research was financially supported in part by the Office of Scientific Research Projects of Van Yuzuncu Yil University under Grant number (TLO-2023-10609).

Project Number

TLO-2023-10609

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