CAJ | 학술논문

目的：探讨鱼腥草挥发油及其单体甲基正壬酮对 LPS-TLR4/ MD-2-TNF-α炎症通路的影响。方法以TLR4/ MD-2阻断剂封闭 TLR4/ MD-2位点，分别运用蛋白免疫印迹法(Western blot)和酶联免疫吸附测定法(ELISA)对鱼腥草挥发油和甲基正壬酮干预后细胞中 TLR4蛋白和炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素10(IL-10)的表达进行检测。同时建立二甲苯致小鼠耳肿胀炎症模型，对鱼腥草挥发油和甲基正壬酮的体内抗炎药效进行比较。结果在1~10μg·mL-1浓度范围内短时干预时，鱼腥草挥发油对 LPS 诱导的 RAW264.7细胞中 TLR4蛋白的抑制作用优于甲基正壬酮，对炎症因子的作用与甲基正壬酮有一定差异。 LPS+TLR4/ MD-2阻断剂+药物干预组与 LPS+TLR4/ MD-2阻断剂组比较，以挥发油干预后细胞中 TLR4蛋白的表达差异无统计学意义(P>0.05)，而以甲基正壬酮干预后细胞中 TLR4蛋白的表达明显减少。表明鱼腥草挥发油通过 LPS-TLR4/ MD-2-TNF-α通路发挥其抗炎作用，而甲基正壬酮可能通过其他途径发挥其抗炎作用。在相同剂量下鱼腥草挥发油的体内抗炎活性优于甲基正壬酮。结论鱼腥草挥发油与甲基正壬酮存在抗炎作用及机制的差异性，应是挥发油中多成分协同作用的结果。
목적：탐토어성초휘발유급기단체갑기정임동대 LPS-TLR4/ MD-2-TNF-α염증통로적영향。방법이TLR4/ MD-2조단제봉폐 TLR4/ MD-2위점，분별운용단백면역인적법(Western blot)화매련면역흡부측정법(ELISA)대어성초휘발유화갑기정임동간예후세포중 TLR4단백화염증인자종류배사인자α(TNF-α)、백세포개소1β(IL-1β)、백세포개소10(IL-10)적표체진행검측。동시건립이갑분치소서이종창염증모형，대어성초휘발유화갑기정임동적체내항염약효진행비교。결과재1~10μg·mL-1농도범위내단시간예시，어성초휘발유대 LPS 유도적 RAW264.7세포중 TLR4단백적억제작용우우갑기정임동，대염증인자적작용여갑기정임동유일정차이。 LPS+TLR4/ MD-2조단제+약물간예조여 LPS+TLR4/ MD-2조단제조비교，이휘발유간예후세포중 TLR4단백적표체차이무통계학의의(P>0.05)，이이갑기정임동간예후세포중 TLR4단백적표체명현감소。표명어성초휘발유통과 LPS-TLR4/ MD-2-TNF-α통로발휘기항염작용，이갑기정임동가능통과기타도경발휘기항염작용。재상동제량하어성초휘발유적체내항염활성우우갑기정임동。결론어성초휘발유여갑기정임동존재항염작용급궤제적차이성，응시휘발유중다성분협동작용적결과。
Objective To explore the effects of volatile oil and 2-undecanone from Houttuynia Cordata Thunb. (H. cordata) on LPS-TLR4 / MD-2-TNF-α signaling pathway. Methods TLR4 / MD-2 blocking agent was used to mask the TLR4 /MD-2 site,then protein expression levels of TLR4 in cells treated with volatile oil and 2-undecanone were analyzed by western blot. ELISA was used to detect the secretion of the inflammatory cytokines such as TNF-α,IL-1β and IL-10. Comparison analysis was then performed from the results of cell experiments in vitro and anti-inflammatory effects through xylene-induced ear edema test in vivo. Results In concentrations between 1 to 10 μg · mL-1 ,Houttuynia volatile oil showed better effect than 2-undecanone on inhibition of TLR4 protein in LPS-induced RAW264. 7 cells, and had some differences in the effects on inflammatory factors. Compared with the LPS+TLR4 / MD-2 group,the LPS+TLR4 / MD-2 + volatile oil group had no significant difference in the expression of TLR4 protein (P>0. 05),but the LPS+TLR4 / MD-2 +2-undecanone group reduced the expression of TLR4 protein obviously. It appeared that volatile oil exerts its anti-inflammatory effect through LPS-TLR4 / MD-2-TNF-αpathway,but 2-undecanone may exert its anti-inflammatory effect by other means. Houttuynia volatile oil showed better anti-inflammatory activity than 2-undecanone in vivo at the same dose. Conclusion There are some differences in anti-inflammatory effects and related mechanisms between volatile oil and 2-undecanone, probably owing to the synergistic effects of multi-ingredients in the volatile oil.