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Inhibition of Pro-inflammatory Mediators and Cytokines by Chlorella Vulgaris Extracts

机译：小球藻提取物对促炎性介质和细胞因子的抑制作用

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Objective: The aim of this study was to determine the in vitro anti-inflammatory activities of solvent fractions from Chlorella vulgaris by inhibiting the production of pro-inflammatory mediators and cytokines. Methods: Methanolic extracts (80%) of C. vulgaris were prepared and partitioned with solvents of increasing polarity viz., n -hexane, chloroform, ethanol, and water. Various concentrations of the fractions were tested for cytotoxicity in RAW 264.7 cells using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and the concentrations inducing cell growth inhibition by about 50% (IC50) were chosen for further studies. Lipopolysaccharide (LPS) stimulated RAW 264.7 cells were treated with varying concentrations of C. vulgaris fractions and examined for its effects on nitric oxide (NO) production by Griess assay. The release of prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6) were quantified using enzyme-linked immunosorbent assay using Celecoxib and polymyxin B as positive controls. Results: MTT assay revealed all the solvent fractions that inhibited cell growth in a dose-dependent manner. Of all the extracts, 80% methanolic extract exhibited the strongest anti-inflammatory activity by inhibiting NO production ( P 2 ( P 2, TNF-α, and IL-6 in LPS activated RAW 264.7 cells. Abbreviations Used: COX-2: Cyclooxygenase-2, DMSO: Dimethyl sulfoxide, FBS: Fetal bovine serum, IL-6: Interleukin 6, iNOS: Inducible nitric oxide synthase, L-NMMA: NG-methyl-L-arginine acetate salt, LPS: Lipopolysaccharide, MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, NO: Nitric oxide, PBS: Phosphate buffered saline, PGE2: Prostaglandin E2, TNF-α: Tumor necrosis factor-α.
机译：目的：本研究的目的是通过抑制促炎性介质和细胞因子的产生来确定小球藻溶剂级分的体外抗炎活性。方法：制备甲醇提取物（80％），并用极性增加的溶剂（正己烷，氯仿，乙醇和水）分配。使用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑（MTT）分析测试了各种浓度的馏分在RAW 264.7细胞中的细胞毒性，并通过约50的浓度诱导了细胞生长选择％（IC 50 ）进行进一步研究。脂多糖（LPS）刺激的RAW 264.7细胞用不同浓度的寻常梭状芽孢杆菌组分处理，并通过Griess分析检查其对一氧化氮（NO）产生的影响。用酶联定量法测定前列腺素E 2 （PGE 2 ），肿瘤坏死因子-α（TNF-α）和白介素6（IL-6）的释放。使用Celecoxib和多粘菌素B作为阳性对照进行免疫吸附测定。结果：MTT分析显示了所有以剂量依赖性方式抑制细胞生长的溶剂组分。在所有提取物中，80％的甲醇提取物通过抑制LPS激活的RAW 264.7细胞中的NO产生（P 2 （P 2 ，TNF-α和IL-6）而表现出最强的抗炎活性。使用的缩写：COX-2：环氧合酶-2，DMSO：二甲基亚砜，FBS：胎牛血清，IL-6：白介素6，iNOS：诱导型一氧化氮合酶，L-NMMA：NG-甲基-L-精氨酸乙酸盐， LPS：脂多糖，MTT：3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑，NO：一氧化氮，PBS：磷酸盐缓冲液，PGE 2 ：前列腺素E 2 ，TNF-α：肿瘤坏死因子-α。

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