Jinhe KANG, Tan YANG, Zhaoliang WU, Jing YANG, Min XIONG, Zhiliang TAN, Zhihong SUN, Wenjun XIAO

Effect of Grape Seed Extract on the Oxidative and Proliferative Status of Porcine Intestinal Epithelial Cells

The aim of this study was to investigate the oxidative and proliferative effects of grape seed extract (GSE). Piglet intestinal epithelial cells (IPEC1) were selected as an unstressed cell model, or they were exposed to 400 μM H2O2 to establish a H2O2-stimulated cell model. The glutathione (GSH) and total antioxidant capacity in response to GSE addition were tested in the unstressed and H2O2-stimulated cell models. The relative mRNA levels of antioxidant or antioxidant enzymes and apoptosis-related genes were measured by Real-Time RT-PCR. In the unstressed status, the cell survival ratio and GSH increased with the addition of GSE at 1 and 10 μg/mL but diminished at 60 μg GSE/mL. The addition of 1 μg/mL GSE upregulated the mRNA expression levels of B-Cell Lymphoma protein-2 (Bcl-2), cysteine aspartases-3 (Caspase-3), cysteine aspartases-8 (Caspase-8) and glutathione peroxidase-1(GPx-1), while it downregulated that of Bcl2-associated X protein (Bax), copper-zinc superoxide dismutase, glutathione S-transferase (GST), thioredoxin, thioltransferase and thioredoxin reductase. As GSE reached 60 μg/mL, the tumor protein p53 (p53) and caspase-8 gene expressions were upregulated. In stressed status, 1 and 10 μg GSE/mL promoted the increase of GSH. H2O2- induced increases in Bax, p53, and Caspase-3 mRNA expressions were attenuated by the subsequent addition of 1 μg GSE/mL and promoted the gene expression of tumor necrosis factor-α, GPx-1 and thioltransferase (Ttas). Treatment with 60 μg GSE/ml resulted in a significant reduction in Bax, p53, manganese superoxide dismutase and GST mRNA expressions. These results indicate that GSE exhibits antioxidative and proliferative functions on unstressed IPEC1 cells at low and medium levels and oxidative and antiproliferative functions at high levels

Üzüm Çekirdeği Ekstraktının Domuz Barsak Epitel Hücreleri Üzerine Oksidatif ve Proliferatif Etkileri

Bu çalışmanın amacı üzüm çekirdeği ekstraktının (ÜÇE) oksidatif ve proliferatif etkilerini araştırmaktır. Domuz barsak epitel hücreleri (IPEC1) strese maruz kalmamış veya 400 μM H2O2 uygulanarak H2O2 ile uyarılmış modeller olarak kullanıldı. ÜÇE ilavesinde strese maruz kalmayan ve H2O2 ile uyarılmış modellerde glutatyon (GSH) ve total antioksidan kapasite tespit edildi. Antioksidanlar veya antioksidan enzimler ile apoptozisle ilişkili genlerin orantısal mRNA seviyesi gerçek zamanlı RT-PCR ile ölçüldü. Strese maruz kalmayanlarda hücre hayatta kalma oranı ve GSH seviyesi 1 ve 10 μg/mL miktarlarında ÜÇE ilavesi ile artarken 60 μg/mL ile azalma gösterdi. 1 μg/mL ÜÇE ilavesi; B-hücre Lenfoma protein-2 (Bcl-2), sistein aspartaz-3 (Kaspaz-3), sistein aspartaz-8 (Kaspaz-8) ve glutatyon peroksidaz-1 (GPx-1) mRNA ekspresyonlarında upregulasyona neden olurken Bcl2-alakalı X protein (Bax), Bakır-çinko süperoksit dismutaz, glutatyon S-transferaz (GST), thioredoksin, thioltransferaz ve thioredoksin reduktaz mRNA ekspresyonlarında downregulasyona neden oldu. 60 μg/mL ÜÇE; tümör protein p53 (p53) ve kaspaz-8 gen ekspresyonunda upregulasyona yol açtı. Strese maruz kalınmada 1 ve 10 μg/mL miktarlarında ÜÇE; GSH miktarındaki artmayı destekledi. H2O2 ile oluşturulan Bax, p53 ve kaspaz-3 mRNA ekspresyonlarındaki artmalar 1 μg/mL ÜÇE ilavesi ile düşürülürken tümör nekroz faktör-α, GPx-1 ve thioltransferaz (Ttas) gen ekspresyonları arttı. 60 μg/mL ÜÇE uygulaması Bax, p53, manganez süperoksit dismutaz ve GST mRNA ekspresyonlarında anlamlı oranda azalmaya neden oldu. Bu sonuçlar; strese maruz kalmamış olan IPEC1 hücrelerinde düşük ve orta seviyede dozlarda ÜÇE uygulamasının antioksidatif ve proliferatif, yüksek dozda ise oksidatif ve antiproliferatif etkileri olduğunu göstermektedir.

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