Expression of Vitamin D Receptor and Vitamin D Receptor Gene Polymorphisms (BsmI, FokI, and TaqI) in Patients with Pterygium / Expressão do gene do receptor da vitamina D e de seus polimorfismos (BsmI, FokI e TaqI) em pacientes com pterígio

ABSTRACT Purpose: This study aimed to determine the role of vitamin D receptor in the pathogenesis of pterygium. The vitamin D receptor eexpression levels in pterygium tissue, blood vitamin D levels, and frequency of selected vitamin D receptor gene polymorphisms (BsmI, FokI, and TaqI) were compared between patients with pterygium and healthy participants. Methods: The study included patients with pterygiumeee (n=50) and healthy volunteers (n=50). The serum vitamin D levels were measured for both groups. Immunohistochemical staining for vitamin D receptor ewas performed on sections obtained from the pterygium and adjacent healthy conjunctival tissues of the same individuals. The genomic existence of vitamin D receptor epolymorphisms (BsmI, FokI, and TaqI) were analyzed in DNA obtained from venous blood of participants using polymerase chain reaction and restriction fragment length polymorphism methods. Results: There was no difference found between the serum vitamin D levels of patients with pterygium and healthy controls. However, tissue expression of vitamin D receptor was higher in the pterygium endothelial cells of micro-vessels (p=0.002), subepithelial stromal (p=0.04), and intravascular inflammatory cells (p=0.0001), in comparison with the adjacent healthy conjunctival tissue. Moreover, while the BBtt haplotype was 2-fold higher, the bbTt haplotype was 2.5-fold lower, and the BbTT haplotype was 2.25-fold lower in the control group than in the pterygium group (p<0.001). Conclusions: Vitamin D serum levels did not differ between the healthy and pterygium groups. Vitamin D receptor expression was increased in the pterygium tissue versus the adjacent healthy tissue. However, vitamin D receptor polymorphism analysis in patients with pterygium did not reveal any significant difference in BsmI, FokI, or TaqI polymorphisms in comparison with the healthy volunteers.(AU)

RESUMO Objetivo: Determinar o papel do receptor da vitamina D na patogênese do pterígio. Os níveis de expressão do receptor da vitamina D no tecido do pterígio, os níveis sanguíneos de vitamina D e a frequência de alguns polimorfismos do gene do receptor da vitamina D (BsmI, FokI e TaqI) foram comparados entre pacientes com pterígio e participantes saudáveis. Métodos: Foram incluídos pacientes com pterígio (n=50) e voluntários saudáveis (n=50). Os níveis séricos de vitamina D foram medidos em ambos os grupos. Foi feita uma coloração imuno-histoquímica para o receptor da vitamina D em cortes obtidos do pterígio e dos tecidos conjuntivais saudáveis adjacentes dos mesmos indivíduos. A existência de polimorfismos do receptor da vitamina D (BsmI, FokI e TaqI) no genoma foi analisada em DNA obtido do sangue venoso dos participantes, usando métodos de Polymerase chain reaction (PCR) e RFLP. Resultados: Não foi observada nenhuma diferença entre os níveis séricos de vitamina D dos pacientes com pterígio e os dos controles saudáveis. Entretanto, a expressão tissular do receptor da vitamina D foi maior nas células endoteliais dos microvasos do pterígio (p=0,002), nas células estromais sub-epiteliais (p=0,04) e nas células inflamatórias intravasculares (p=0,0001), quando comparada à expressão no tecido conjuntival saudável adjacente. Além disso, embora o haplótipo BBtt tenha sido duas vezes mais frequente, o haplótipo bbTt foi 2,5 vezes menos frequente e o haplótipo BbTT foi 2,25 vezes menos frequente no grupo de controle do que no grupo com pterígio (p<0,001). Conclusões: Os níveis séricos de vitamina D não apresentaram diferenças entre o grupo de pessoas saudáveis e o com pterígio. A expressão do receptor da vitamina D mostrou-se maior no grupo com pterígio do que no tecido saudável adjacente. Entretanto, a análise dos polimorfismos do receptor da vitamina D nos pacientes com pterígio não revelou qualquer diferença significativa nos polimorfismos BsmI, FokI ou TaqI em comparação com os voluntários saudáveis.(AU)

Cell viability, anti-proliferation and antioxidant activities of Sideritis syriaca, Tanacetum argenteum sub sp. argenteum and Achillea aleppica subsp. zederbaueri on human breast cancer cell line (MCF-7).

The breast cancer is one of the most common types of cancer. Many scientists have focused on the treatment of this disease for a long time by studying anti-cancer activities of plant extracts as well as synthetics. Lamiaceae and Asteraceae have been used in anticancer studies due to their phytochemical content. The genus Sideritis, Achillea and Tanacetumare the members of these families. Sideritis, Achillea and Tanacetum are used as herbal medication for the treatment ofvariety of diseases. In present study, we demonstrated the biological activity of Sideritis syriaca (SS), Achillea aleppica (AAZ) and Tanacetum argenteum (TAA) methanol extracts on cell viability of the breast cancer line MCF7. MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to determine the cell viability and proliferation of MCF7 cells. In a dose dependent manner, methanol extracts (0, 1, 5, 25, 100 and 250 μg/ml) of SS, AZZ and TAA were examined on MCF7, and viability of cells were determined with MTT staining. Especially, concentrations in 100 and 250 μg/ml of extracts decreased the cell viability (p<0.001). The results of the current study showed that methanol extracts of SS, AZZ and TAA effectively inhibited the cell proliferation by decreasing the cell viability of MCF7 cells. Suggesting that SS, AZZ and TAA can be considered as natural herbal-based anti-cancerous agents.