School of Medical Science and Technology, Indian Institute of Technology Kharagpur , Kharagpur 721302, West Bengal, India.
Department of Biotechnology, National Institute of Technology Sikkim , Barfung Block Ravangla Sub-Division South Sikkim, Ravangla 737139, Sikkim, India.
J Agric Food Chem. 2017 May 3;65(17):3460-3473. doi: 10.1021/acs.jafc.7b00366. Epub 2017 Apr 20.
Hypoxic assault affects fundamental cellular processes and generates oxidative stress on healthy cells/molecules. Honey extracted polyphenolics (HEP) as a natural antioxidant reduced hypoxic cytotoxicity in this study. Different honey samples were physicochemically characterized to identify preferred (jamun) honey [pH 3.55 ± 0.04, conductivity (μs/cm) = 6.66 ± 0.14, water content % (w/w) = 14.70 ± 0.35, total solid content % (w/w) = 85.30 ± 0.35, phenol content (mg GAE/100 g) = 403.55 ± 0.35, flavonoid content (mg QE/100 g) = 276.76 ± 4.10, radical scavenging activity (% 500 μL) = 147.75 ± 3.13, catalase activity (absorbance at 620 nm) = 0.226 ± 0.01]. HEP was tested in different doses on hypoxic and normoxic cells (HaCaT) using viability and antioxidant assays. Cardinal molecular expressions such as cadherin-catenin-cytoskeleton complex (namely, E-cadherin, β-catenin, and F-actin), hypoxia marker (Hif 1 α), proliferation marker (Ki67), and epithelial master regulator (p63) were studied by immuno-cytochemisty (ICC) and qRT-PCR. The 0.063 mg/mL HEP demonstrated better vitality and functionality of HaCaT cells as per viability assay (*, P < 0.01) even under hypoxia. ICC and qRT-PCR observations indicated restoration of cellular survival and homeostasis under 0.063 mg/mL HEP after hypoxic assault. Furthermore, major spectral changes for nucleic acid and membrane phospholipid reorganizations by Fourier transform infrared spectroscopy illustrated a positive impact of 0.063 mg/mL HEP on hypoxic cells considering proliferation and cellular integrity. It was concluded that a specific dose of jamun HEP reduces hypoxic cytotoxicity.
缺氧攻击会影响健康细胞/分子的基本细胞过程并产生氧化应激。在这项研究中，蜂蜜中提取的多酚（HEP）作为一种天然抗氧化剂，降低了缺氧的细胞毒性。对不同的蜂蜜样本进行了物理化学特性分析，以确定首选的（黑刺李）蜂蜜[pH3.55±0.04，电导率（μs/cm）=6.66±0.14，水含量%（w/w）=14.70±0.35，总固体含量%（w/w）=85.30±0.35，酚含量（mgGAE/100g）=403.55±0.35，类黄酮含量（mgQE/100g）=276.76±4.10，自由基清除活性（%500μL）=147.75±3.13，过氧化氢酶活性（在 620nm 处的吸光度）=0.226±0.01]。使用细胞活力和抗氧化测定法，在不同剂量下将 HEP 测试于缺氧和常氧细胞（HaCaT）上。通过免疫细胞化学（ICC）和 qRT-PCR 研究了主要分子表达，如钙粘蛋白-连环蛋白-细胞骨架复合物（即 E-钙粘蛋白、β-连环蛋白和 F-肌动蛋白）、缺氧标志物（Hif1α）、增殖标志物（Ki67）和上皮主调控因子（p63）。根据细胞活力测定法（*，P<0.01），0.063mg/mLHEP 显示出更好的 HaCaT 细胞活力和功能，即使在缺氧条件下也是如此。ICC 和 qRT-PCR 观察表明，在 0.063mg/mLHEP 缺氧攻击后，细胞存活和体内平衡得到恢复。此外，傅里叶变换红外光谱的核酸和膜磷脂重排的主要光谱变化表明，考虑到增殖和细胞完整性，0.063mg/mLHEP 对缺氧细胞有积极影响。研究得出结论，特定剂量的黑刺李 HEP 可降低缺氧的细胞毒性。
