威灵仙不同提取部位对脂多糖诱导的RAW264.7细胞NO生成及iNOS和TNF-α表达的影响

Effect of Different Fractions Extracted fromClematis Chinensis Osbeck on NO,iNOS and TNF-αExpressions in Lipopolysaccharide Induced RAW264.7Cells

研究了威灵仙不同提取部位对脂多糖(LPS)诱导的RAW264.7细胞NO生成及iNOS和TNF-α分泌的作用.通过ELISA方法检测25、50、100μg/mL的威灵仙不同提取部位对LPS(1μg/mL)诱导RAW264.7细胞产生NO、iNOS和TNF-α的影响;实时定量PCR方法检测威灵仙不同提取部位对LPS诱导iNOS和TNF-α基因表达的影响.结果表明,LPS能够明显诱导RAW264.7细胞产生NO、iNOS和TNF-α,威灵仙不同提取部位(25、50、100μg/mL)均能抑制其活性和基因表达,并呈现明显的剂量依赖性,其作用强弱依次为威灵仙乙酸乙酯部位、石油醚部位、正丁醇部位和水部位.因此得出,威灵仙不同提取部位的抗炎作用效果不同;抑制炎症因子的分泌和释放可能是威灵仙发挥抗炎作用机制之一.

We studied the different fractions extracted from Clematis Chinensis Osbeck on NO,iNOS and TNF-α expressions in lipopolysaccharide （LPS） induced RAW264.7 cells.The contents of NO,iNOS and TNF-α were detected by ELISA methods; the mRNA expression levels of iNOS and TNF-α were analyzed by real-time PCR in presence or absence of 1 LPS （1μg/mL） or different fractions extracted from Clematis Chinensis Osbeck （25、50、100 μg/mL）.In the study,we found that NO,iNOS and TNF-α could be induced from RAW264.7 cells induced by LPS.Different fractions extracted from Clematis Chinensis Osbeck （25、50、100 μg/ mL） inhibited their activities and gene expressions in dose dependent manners,and the effects from strong to weak are as follows：the ethyl acetate fraction,petroleum ether fraction,n-butyl alcohol fraction and water fraction.Anti-inflammatory effects of different fractions extracted from Clematis Chinensis Osbeck were different,mainly inhibiting the secretion and release of the mediators of inflammation and inflammatory factor.