Publication: UVB ışınına maruz bırakılmış farklı mantar türlerinin insan osteoblast hücre hattında kemik metabolizmasına etkisi
Abstract
Amaç: Farklı dozlarda UVB uygulanmış ve UVB uygulanmamış mantarların [Kültür mantarı (Agaricus bisporus), Shiitake mantarı (Lentinula edodes) ve Kral mantarı (Pleurotus eryngii)] insan osteoblast hücre hattında kemik metabolizması üzerine etkilerini araştırmak ve bu mantarların kemik metabolizması üzerine etkilerini D2 ve D3 vitaminleri ile karşılaştırmaktır. Gereç ve Yöntem: Numunelerdeki D2 vitamini seviyeleri HPLC kullanılarak belirlenmiştir. Antioksidan aktivitesini değerlendirmek için CUPRAC, DPPH ve FRAP analizleri yapılmış ve numunelerin toplam fenolik bileşen miktarı ölçülmüştür. hFOB 1.19 hücrelerinin canlılığını belirlemek için MTT testi kullanılmıştır. Numunelerin hFOB 1.19 hücrelerindeki ALP ve OPG seviyeleri üzerindeki etkisini değerlendirmek için ELISA yöntemi; BGLAP, RANKL ve RUNX2 ekspresyonlarına etkisini ölçmek için gerçek zamanlı PCR yöntemi uygulanmıştır. Hücre morfolojisinin değerlendirilmesi için konfokal mikroskopi kullanılmıştır. Bulgular: UVB uygulamasının, mantarların D2 vitamini seviyelerini, antioksidan kapasitesini ve fenolik bileşen içeriğini artırdığı gözlemlenmiştir. İnsan osteoblast hücrelerinde ise canlılık oranının, ALP ve OPG seviyelerinin mantarlara UVB uygulanmasına bağlı olarak yükseldiği saptanmıştır. Ayrıca, Agaricus bisporus’a UVB uygulanması ve UVB dozunun artmasıyla, insan osteoblast hücrelerinde BGLAP ve RUNX2 ekspresyonunun arttığı, RANKL ekspresyonunun ise azaldığı tespit edilmiştir. Bunun yanı sıra, mantarlara uygulanan UVB dozunun artmasıyla insan osteoblast hücrelerinin konfokal görüntülerinde hücre yoğunluğunun görsel olarak arttığı gözlenmiştir. Sonuç: Mantarlara UVB uygulanması kemik biyobelirteçlerini olumlu yönde etkilemiştir. Bu nedenle, UVB uygulanmış mantarların kemik sağlığını korumada potansiyel bir doğal tedavi stratejisi olarak değerlendirilebileceği düşünülmektedir.
Objective: To investigate the effects of different doses of UVB-applied and non-UVB-applied mushrooms [White button mushroom (Agaricus bisporus), Shiitake mushroom (Lentinula edodes) and King oyster mushroom (Pleurotus eryngii)] on bone metabolism in human osteoblast cell line and to compare the effects of these mushrooms on bone metabolism with vitamins D2 and D3. Material and Method: The levels of vitamin D2 in the samples were determined using HPLC. To evaluate antioxidant activity, CUPRAC, DPPH, and FRAP assays were performed, and the total phenolic content of the samples was measured. The MTT assay was used to determine the viability of hFOB 1.19 cells. The ELISA method was employed to evaluate the effects of the samples on ALP and OPG levels in hFOB 1.19 cells, while real-time PCR was used to assess their impact on BGLAP, RANKL, and RUNX2 expression. Confocal microscopy was used to evaluate cell morphology. Results: It has been observed that UVB application increases the levels of vitamin D2, antioxidant capacity, and phenolic content in mushrooms. In human osteoblast cells, it was found that cell viability, as well as ALP and OPG levels, increased in response to UVB application to the mushrooms. Furthermore, with the application of UVB to Agaricus bisporus and the increase in UVB dose, BGLAP and RUNX2 expression in human osteoblast cells increased, while RANKL expression decreased. Additionally, a visual increase in cell density was observed in the confocal images of human osteoblast cells as the UVB dose applied to the mushrooms increased. Conclusion: UVB application to mushrooms has positively affected bone biomarkers. Therefore, it is thought that UVB-applied mushrooms could be considered a potential natural treatment strategy for maintaining bone health.
Objective: To investigate the effects of different doses of UVB-applied and non-UVB-applied mushrooms [White button mushroom (Agaricus bisporus), Shiitake mushroom (Lentinula edodes) and King oyster mushroom (Pleurotus eryngii)] on bone metabolism in human osteoblast cell line and to compare the effects of these mushrooms on bone metabolism with vitamins D2 and D3. Material and Method: The levels of vitamin D2 in the samples were determined using HPLC. To evaluate antioxidant activity, CUPRAC, DPPH, and FRAP assays were performed, and the total phenolic content of the samples was measured. The MTT assay was used to determine the viability of hFOB 1.19 cells. The ELISA method was employed to evaluate the effects of the samples on ALP and OPG levels in hFOB 1.19 cells, while real-time PCR was used to assess their impact on BGLAP, RANKL, and RUNX2 expression. Confocal microscopy was used to evaluate cell morphology. Results: It has been observed that UVB application increases the levels of vitamin D2, antioxidant capacity, and phenolic content in mushrooms. In human osteoblast cells, it was found that cell viability, as well as ALP and OPG levels, increased in response to UVB application to the mushrooms. Furthermore, with the application of UVB to Agaricus bisporus and the increase in UVB dose, BGLAP and RUNX2 expression in human osteoblast cells increased, while RANKL expression decreased. Additionally, a visual increase in cell density was observed in the confocal images of human osteoblast cells as the UVB dose applied to the mushrooms increased. Conclusion: UVB application to mushrooms has positively affected bone biomarkers. Therefore, it is thought that UVB-applied mushrooms could be considered a potential natural treatment strategy for maintaining bone health.