Immunology and Microbiology Laboratory, Department of Human Physiology, Community Health, Vidyasagar University, Midnapore, West Bengal, India.
Oxid Med Cell Longev. 2009 Sep-Oct;2(4):222-30. doi: 10.4161/oxim.2.4.9000.
In the present study, methanol extract of Ocimum gratissimum Linn (ME-Og) was tested against nicotine-induced murine peritoneal macrophage in vitro. Phytochemical analysis of ME-Og shown high amount of flavonoid and phenolic compound present in it. The cytotoxic effect of ME-Og was studied in murine peritoneal macrophages at different concentrations (0.1 to 100 microg/ml) using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide (MTT) method. To establish the protective role of ME-Og against nicotine toxicity, peritoneal macrophages from mice were treated with nicotine (10 mM), nicotine+ME-Og (1 to 25 microg/ml) for 12h in culture media. The significantly (P< 0.05) increased super oxide anion generation, reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity, myeloperoxidase (MPO) activity, lipid peroxidation, protein carbonyls, oxidized glutathione levels were observed in nicotine-treated group as compared to control group; those were significantly (p < 0.05) reduced in ME-Og supplemented groups in concentration dependent manner. More over, significantly (p < 0.05) reduced antioxidant status due to nicotine exposure was effectively ameliorated by ME-Og supplementation in murine peritoneal macrophages. Among the different concentration of ME-Og, maximum protective effect was observed by 25 microg/ml, which does not produce significant cell cytotoxicity in murine peritoneal macrophages. These findings suggest the potential use and beneficial role of O. gratissimum as a modulator of nicotine-induced free radical generation, lipid-protein damage and antioxidant status in important immune cell, peritoneal macrophages.
在本研究中,我们测试了罗勒甲醇提取物(ME-Og)对尼古丁诱导的小鼠腹腔巨噬细胞的体外作用。ME-Og 的植物化学成分分析表明,其中含有大量的类黄酮和酚类化合物。我们采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法,在不同浓度(0.1 至 100μg/ml)下研究 ME-Og 对小鼠腹腔巨噬细胞的细胞毒性作用。为了确定 ME-Og 对尼古丁毒性的保护作用,我们用尼古丁(10mM)处理小鼠腹腔巨噬细胞,在培养物中与 ME-Og(1 至 25μg/ml)共同孵育 12 小时。与对照组相比,在尼古丁处理组中观察到超氧阴离子生成显著增加(P<0.05)、烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶活性、髓过氧化物酶(MPO)活性、脂质过氧化、蛋白羰基、氧化型谷胱甘肽水平降低;这些变化在 ME-Og 补充组中呈浓度依赖性降低。此外,由于尼古丁暴露导致的抗氧化状态显著降低,通过 ME-Og 补充得到了有效改善。在不同浓度的 ME-Og 中,25μg/ml 的效果最佳,在小鼠腹腔巨噬细胞中没有产生显著的细胞毒性。这些发现表明,罗勒具有作为尼古丁诱导的自由基生成、脂质-蛋白质损伤和重要免疫细胞——腹腔巨噬细胞中抗氧化状态调节剂的潜在用途和有益作用。