CC-K8抑制LPS诱导的大鼠肺间质巨噬细胞TLR4及IL-1β的表达

Inhibitory effect of CCK-8 on the expressions of TLR4 and IL-1β in LPS-induced pulmonary interstitial macrophages of rats

目的观察八肽胆囊收缩素(cholecystokininoctapeptide ,CCK 8)对外源性脂多糖(lipopolysaccharide ,LPS)激活大鼠肺间质巨噬细胞(pulmonaryinterstitialmacrophages,PIMs)Toll样受体4 (Tolllikereceptor4 ,TLR4 )及IL 1β表达的影响,探讨CCK- 8的抗炎作用机制。方法分离培养大鼠PIMs ,经LPS、CCK- 8及溶剂单独或共同孵育不同时间后,采用Northernblot、ELISA、RT PCR技术检测TLR4mRNA、IL- 1β及IL- 1βmRNA表达的变化。结果LPS(1mg/L)刺激可使PIMs中TLR4mRNA表达明显增强;随着LPS孵育时间的延长,细胞中的IL -1β含量逐渐增多,2 4h达高峰,IL- 1βmRNA表达水平同时明显升高;CCK 8可剂量依赖性抑制LPS诱导的大鼠PIMsTLR4mRNA、IL- 1β及IL 1βmRNA的表达。结论CCK 8对LPS激活的PIMsTLR4及IL- 1β表达有负性调节作用,是CCK- 8抗炎作用机制之一。

In order to study the effect of cholecystokinin octapeptide(CCK-8)of inhibiting the expression of TLR4 and IL-1β in rat pulmonary interstitial macrophages (PIMs) induced by lipopolysaccharide. Methods PIMs was isolated and cultured in the presence or absence of LPS, CCK-8, and vehicle. The expression of TLR4 mRNA and IL-1β mRNA was assayed by Northern blot and RT-PCR. IL-1β in the supernatant of culture media was detected by ELISA. Results TLR4 mRNA and IL-1β mRNA in rat PIMs were up-regulated by LPS(1mg/L). IL-1β production were increased by LPS incubation in a time-dependent manner. CCK-8, at concentrations of 10 -7 and 10 -6 mol/L, significantly inhibited the production of IL-1β and the expression of TLR4 mRNA and IL-1β mRNA. Conclusion The results indicated that CCK-8 negatively modulated the expression of TLR4 and IL-1β in rat PIMs induced by LPS. This may be one of the mechanisms for CCK-8 to alleviate inflammation in lung tissue during endotoxemia.