摘要
目的:观察VOHS(荆芥挥发油)、VOCC(桂枝挥发油)体外干预小鼠腹腔巨噬细胞TLR2/4通路的抗炎机制。方法:1.常规培养小鼠腹腔巨噬细胞,加入不同浓度的VOHS、VOCC,药物作用24 h后采用MTT法检测细胞存活率。2.常规培养小鼠腹腔巨噬细胞,分组加药培养24 h后,提取细胞总RNA,采用RT-PCR测定TLR2、TLR4 mRNA表达。3.常规培养小鼠腹腔巨噬细胞,分组加药培养6h后,收集细胞上清液,采用ELISA测定其中TNF-α。结果:1.VOHS和VOCC在0.00625～0.025 mg/mL剂量范围内,细胞存活率与对照组相比无差异(P>0.05),可作为受试剂量。2.VOHS、VOCC在0.025 mg/mL剂量下能显著降低LPS所致的小鼠巨噬细胞TLR2/4 mRNA表达的过度升高(P﹤0.05)。3.VOHS在0.025 mg/mL、0.0125 mg/mL剂量下和VOCC在0.025 mg/mL剂量下能显著抑制LPS刺激的小鼠腹腔巨噬细胞对TNF-α的分泌与释放(P<0.05)。结论:VOHS、VOCC可明显抑制LPS诱导的小鼠腹腔巨噬细胞TLR2、TLR4mRNA高表达,及减少前炎症细胞因子TNF-α含量的释放,这可能荆芥和桂枝的抗炎机制之一。
Objective：To study the effect of VOHS and VOCC on the expression of mRNA of TLR2/4 of mouse peritoneal macrophage which stimulated by LPS in vitro.Methods： ①Mouse peritoneal macrophage was cultivated by routine,VOHS and VOCC were used to treat peritoneal macrophage with different dosages in vitro,and then the rate of cell survival was detected by MTT at 24 h respectively.②Mouse peritoneal macrophage were cultivated by routine,VOHS and VOCC were used to treat the peritoneal macrophages which stimulated by LPS in vitro,then the expression TLR2/4 of mRNA were detected by RT-PCR at 24h point.③Mouse peritoneal macrophage were cultivated by routine,VOHS and VOCC were used to treat the peritoneal macrophages which stimulated by LPS in vitro,then the levels of TNF-α in clear supernatant liquid were detected by ELISA at 6h point.Results： ①Within the dosage of 0.00625~0.025 mg/mL of VOHS and VOCC,the rates of cell survival of VOHS＇s group and VOCC＇s group are same with the control group（P0.05）,so the dosage of test was determined.②The expression of TLR2/4 of mRNA in peritoneal macrophage which stimulated by LPS was decreased at the dosage of 0.025mg/ml of VOHS and VOCC（P0.05）③The level of TNF-α in clear supernatant liquid was decreased at the dosage of 0.025 mg/mL,0.0125 mg/mL of VOHS and 0.025 mg/mL of VOCC（P0.05）.Conclusion： The expression of TLR2/4 of mRNA in peritoneal macrophage which stimulated by LPS was restrained by VOHS and VOCC,and also the level of TNF-α.This maybe is one of anti-inflammatory mechanisms of Schizonepetae and Cinnamomun.
出处
《成都中医药大学学报》
2011年第2期56-60,共5页
Journal of Chengdu University of Traditional Chinese Medicine
基金
国家自然基金资助项目(编号:30472201)
关键词
荆芥挥发油
桂枝挥发油
脂多糖
腹腔巨噬细胞
Toll样受体2/4
基因表达
Volatile oil of herba schizonepetae
VOHS
volatile oil of cinnamomun Cassia
VOCC
peritoneal macrophage
toll-like receptors 2/4
TLR2/4
gene expression
