摘要
目的:观察泻心汤有效组分结合蒽醌与总黄酮配伍(重量比为4∶3)对LPS诱导巨噬细胞NO合成以及对iNOS、CD14和TLR4 mRNA表达的影响。方法:采用MTT法观察结合蒽醌与总黄酮配伍对巨噬细胞生长活性的影响,采用G riess法观察其对LPS诱导的巨噬细胞产生NO的影响,采用RT-PCR法观察其对巨噬细胞iNOS、CD14和TLR4 mRNA表达在2、4、6、24h的影响。结果:泻心汤结合蒽醌与总黄酮配伍在0.01-0.1 mg/m l剂量对巨噬细胞生长活性无明显影响,此浓度能明显抑制LPS诱导的巨噬细胞NO分泌,结合蒽醌与总黄酮配伍组TLR4和CD14 mRNA表达在2h明显低于LPS模型组,而iNOS mRNA表达与模型组无明显差异,4h以后iNOS和TLR4 mRNA的表达均明显低于模型组,而CD14 mRNA表达在4h后与模型组无明显差异。结论:泻心汤结合蒽醌与总黄酮配伍抑制LPS诱导的巨噬细胞NO合成、iNOS mRNA表达与其抑制TLR4 mRNA的表达密切相关,而与CD14的关系不大,TLR4可能是泻心汤结合蒽醌与总黄酮配伍抗内毒素的作用靶点。
Objective: To observe the effect of compatibility of Conjunct anthraquinone and total flavonoids (proportion by weight is 4:3 ) on NO production, and expression of iNOS,CD14 and TLR4mRNA in peritoneal macrophages induced by LPS. Methods: Growth of macrophages cultured with different levels of the compatibility was determined by MTT, NO production in macrophages induced by LPS was detected by Griess method, the expression of iNOS,CD14 and TLR4mRNA in macrophages at the time of 2,4,6 and 24h induced by LPS were detected by RT- PCR method. Results: Therewas no obvious influence on growth activity of macrophages at dose of 0. 01 -0.1 mg/ml of the compatibility , the NO level was inhibited obviously by the compatibility at those dose after 4h, the expression of TLR4 and CD14mRNA were inhibited obviously by the compatibility compared with LPS group at 2h( P 〈 0.05 ), but the expression of iNOSmRNA had no obvious difference between the compatibility and LPS group at the same time, the expression of TLR4 and iNOSmRNA were inhibited obviously after 4h( P 〈 0. 05 ) , but the expression of CD14mRNA showed no significant difference between the compatibility and LPS group at the same time after 4h (P 〈0.05). Conclusion: The results suggested that the down - regulation of NO production and expression of iNOS mRNA were related with TLR4, but less with CD14,TLR4 was the possible antiendotoxic target of the compatibility of active components of Xiexin Decoction.
出处
《中药药理与临床》
CAS
CSCD
北大核心
2007年第5期27-30,共4页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金资助项目(No.30371756)