【作者】 宋美芳；

【导师】 曾南；

【作者基本信息】 成都中医药大学 ， 药理学， 2009， 硕士

【摘要】 荆芥(Schizonepeta tenuifolia Briq)具有祛风解表,宣毒透疹,理血止痉之功效,桂枝(Cinnamomun cassia Presl.)具有发汗解表,温经通阳之功效,两者为常用解表药,用于表证的治疗。本课题依据荆芥与桂枝的功效和临床应用,基于两者挥发油具有良好抗炎作用(对急、慢性炎症及免疫损伤性炎症均有良好抑制作用),且体内抗炎作用机制与TLR2/4通路相关的前期研究成果,基于TLR通路在炎症级联反应中的重要地位,以小鼠腹腔巨噬细胞为研究载体,采用脂多糖(LPS)刺激所致的体外细胞炎症模型,应用RT-PCR、酶联免疫的方法观察荆芥挥发油(the volatile oil of Herba Schizonepetae,VOHS)、桂枝挥发油(the volatile oil of Cinnamomun cassia,VOCC)体外给药对LPS刺激的小鼠腹腔巨噬细胞TLR2、TLR4mRNA表达及前炎症细胞因子TNF-α含量的影响,从而达到从体内与体外两方面探讨、阐述荆芥、桂枝挥发油干预TLR信号转导途径抗炎机制的目的。结果如下：1.体外经LPS刺激的小鼠腹腔巨噬细胞TLR2、TLR4mRNA表达过度,且细胞培养上清液中TNF-α含量增加,表明该细胞炎症模型建立成功。2.VOHS、VOCC (0.025mg/ml)能显著降低LPS所致的小鼠腹腔巨噬细胞TLR2、TLR4mRNA表达的过度升高(P<0.05)；0.0125、0.00625mg／ml作用不明显,各剂量之间无明显量效关系。3.VOHS (0.025mg/ml、0.0125 mg/ml)和VOCC (0.025mg/ml)能显著抑制LPS刺激的小鼠腹腔巨噬细胞TNF-α的分泌与释放(P<0.05),其余剂量组作用不明显。综上,VOHS、VOCC抑制LPS诱导的小鼠腹腔巨噬细胞TLR2、TLR4mRNA表达的升高及减少前炎症细胞因子TNF-α含量的释放可能是其体外抗炎机制之一,为两种解表药挥发油的抗炎作用机制的完善提供了一定药理学依据,但有待进一步深入研究。更多还原

【Abstract】 Schizonepeta tenuifolia Briq. has the efficiency of expelling pathogenic wind from the body surface etc, and Cinnamomun cassia Presl. has the efficiency of inducing sweat and dispelling exogenous evils; warming meridian to activate yang.These two kinds of Chinese medicine herbs belong to the herbs of relieving exterior syndromes in common use. According to their efficiency and clinical using, and based on the prior research findings that both of their volatile oils have significant anti-inflammation action (significantly suppress acute; chronic inflammation as well as immunologic injury inflammation), which the mechanism were related to TLR2/4 pathway in vivo. In the present study, mouse peritoneal macrophage were used to establish cell inflammatory model induced by LPS in vitro, to investigate the effect of VOHS, VOCC on TLR2/4mRNA expression and the content of pro-inflammatory cytokines TNF-a in the mouse peritoneal macrophage by RT-PCR, ELISA methods, so as to explain the anti-inflammation mechanism of these two kinds of voliate oil on TLR signal transduction pathway in vitro.1. TLR2/4mRNA expression were excessive high in the mouse peritoneal macrophage induced by LPS in vitro and the content of TNF-a in cultural supernatants increased significantly, that indicated the cell model of inflammatory be established successfully.2. VOHS、VOCC (0.025mg/ml) could reduce evidently the TLR2/4mRNA excessive expression in the mouse peritoneal macrophage induced by LPS(P< 0.05); and VOHS、VOCC (0.0125mg/ml、0.00625mg/ml) had no obvious reducing effects. The dose-effect relationship was not clear among each dosage.3. VOHS(0.025mg/ml、0.0125 mg/ml) and VOCC (0.025mg/ml) couldinhibit markedly secretion and release of TNF-a in the mouse peritoneal macrophage induced by LPS (P<0.05), while no obvious effect were found in the other dosage groups.Taken together, the present results demonstrate that the suppressing effects of VOHS, VOCC on TLR2/4mRNA excessive expression in the mouse peritoneal macrophage induced by LPS and reducing the content release of pro-inflammatory cytokines TNF-a may be one of the anti-inflammation mechanism in vitro, which provide some evidences for the interfering in TLR signal pathway of VOHS, VOCC to exhibit anti-inflammation action. But further studies are need.更多还原