摘要
目的 :观察硫酸化八肽胆囊收缩素 (CCK - 8)对体外脂多糖 (LPS)诱导大鼠肺间质巨噬细胞 (PIMs)TNF -α基因表达的影响 ,探讨核因子κB(NF -κB)是否参与这一过程 ,以揭示CCK - 8抗炎作用的信号转导机制。方法 :分离大鼠肺PIMs ,经LPS、CCK - 8、CCK受体拮抗剂丙谷胺及溶剂单独或联合应用孵育 3h ,用RT -PCR技术检测细胞TNF -αmRNA的表达 ,孵育 1h ,用电泳迁移率改变分析方法检测NF -κB活性 ,孵育 30min ,用Westernblot技术检测胞浆IκBα蛋白表达情况。结果 :CCK - 8(10 -8- 10 -6mol·L-1)明显降低了LPS诱导的TNF -αmRNA表达及NF-κB活性 ,增加了胞浆中IκBα蛋白水平 ,呈剂量依赖性 ,并可被丙谷胺所拮抗。结论 :对LPS激活的肺PIMs ,CCK - 8通过抑制NF -κB活性而抑制其TNF -αmRNA表达 ,该作用由CCK受体介导 ,并与CCK - 8减少IκBα蛋白降解有关 ,此为CCK -
AIM: To elucidate the anti-inflammatory mechanism of cholecystokinin octapeptide (CCK-8). METHODS: The pulmonary interstitial macrophages (PIMs) from rats were stimulated with LPS (1 mg·L^(-1)) in the presence or absence of CCK-8 (10^(-8)-10^(-6) mol·L^(-1)) or/and CCK receptor antagonist proglumide (2 mg·L^(-1)). The expression of TNF-α mRNA was assayed by reverse transcription polymerase chain reaction (RT-PCR) at 3 h of the stimulation, and nuclear factor-κB (NF-κB) binding activity was analyzed by electrophoretic mobility shift assay (EMSA) at 1 h of stimulation. The IκBα protein level in the cytoplasma at 30 min of the stimulation was detected by Western blot. RESULTS: CCK-8, at concentrations from 10^(-8) mol·L^(-1) to 10^(-6) mol·L^(-1) obviously inhibited LPS-induced TNF-α mRNA expression and NF-κB binding activity in a dose-dependent manner. Stimulation with LPS resulted in a reduction of IκBα protein level in PIMs, which was elevated by CCK-8. The effects of CCK-8 on NF-κB activity and IκB protein level were attenuated by CCK receptor antagonist proglumide. CONCLUSION: CCK-8 inhibits LPS-induced TNF-α mRNA expression by regulating NF-κB activity in rat PIMs, which is mediated through CCK receptors and inhibition of IκBα degradation. This represents one of the anti-inflammatory mechanisms of CCK-8.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第8期1335-1339,共5页
Chinese Journal of Pathophysiology
基金
河北省自然科学基金资助项目 (No .30 0 32 2 )
