摘要
目的探讨毛蕊异黄酮对脂多糖(LPS)和γ干扰素(INF-γ)诱导的RAW264.7分泌一氧化氮(NO)的影响。方法LPS和INF-γ共同刺激生长良好的RAW264.7细胞,并用不同浓度(25、50、100μmol/L)毛蕊异黄酮进行干预;MTT法检测毛蕊异黄酮对RAW264.7细胞的毒性作用;硝酸还原酶法检测细胞上清液中NO含量;实时定量PCR法检测细胞中诱生型一氧化氮合酶(iNOS)mRNA的表达水平。结果 125、50、100μmol/L浓度的毛蕊异黄酮对RAW264.7细胞无毒性作用(P>0.05);2LPS和INF-γ共同刺激RAW264.7细胞后,细胞上清液中NO的含量明显增加(P<0.01);毛蕊异黄酮进行干预后,NO的含量明显下降,并呈现出浓度依赖性(P<0.01);3用LPS和INF-γ共同刺激RAW264.7细胞后,细胞中iNOS mRNA的表达水平显著升高(P<0.01);毛蕊异黄酮进行干预后,iNOS mRNA表达水平显著降低,并呈现出浓度依赖性(P<0.01)。结论毛蕊异黄酮能抑制LPS和INF-γ诱导的RAW264.7细胞产生NO,其抗肿瘤作用可能与此有关。
Objective To investigate the effects of calycosin on nitric oxide (NO)secretion in RAW264.7 cells induced by lipopolysaccharide (LPS)and gamma interferon (INF-γ).Methods RAW264.7 cells were induced by LPS and INF-γ,and then different concentrations (25,50,100 μmol /L)of calycosin were added in the culture medium.The cytotoxicity of calycosin was measured with MTT method.The contents of NO in the supernatant were determined with Griess method.Changes in the expression of inducible nitric oxide synthase (iNOS)mRNA were detected with qRT-PCR. Results ① Calycosin showed no cytotoxicity on RAW 264.7 cells at the concentrations of 25,50 and 100 μmol/L (P 〉0.05).② The secretion of NO statistically increased in RAW264.7 cells induced by LPS and INF-γ;Calycosin (25,50,100 μmol/L)exhibited inhibitory effects in a concentration-dependent manner (P 〈0.01).③ The expressions of iNOS mRNA statistically increased in RAW264.7 cells induced by LPS and INF-γ.Calycosin (25,50,100 μmol/L)exhibited inhibitory effects in a concentration - dependent manner (P 〈0.01).Conclusion Calycosin can inhibit NO secretion in LPS and INF-γinduced RAW264.7 cells, which might be associated with the antineoplastic function.
出处
《国外医学(医学地理分册)》
CAS
2015年第1期9-12,共4页
Foreign Medical Sciences:Section of Medgeography
基金
国家自然科学基金资助项目(No.81273196)