摘要
目的:探讨胆囊收缩素(cholecystokinin,CCK)受体CCK—AR及CCK—BR mRNA在大鼠肺间质巨噬细胞(PIMs)内的表达及脂多糖(lipopolysaccharide,LPS)对其表达的影响。方法:用酶消化法结合肺泡灌洗和肺循环灌洗技术分离纯化大鼠PIMs,用LPS(10 mg/L)孵育PIMs 0.5-12 h。采用RT-PCR及Southern印迹技术观察CCK受体在大鼠PIMs内的表达亚型及LPS对其表达的影响。结果:经RT—PCR检测显示。CCK—AR和CCK—BR mRNA在大鼠PIMs均有表达,CCK—ARmRNA RT—PCR扩增产物为1.37 kb,CCK—BR mRNA RT—PCR扩增产物为480 bp。CCK—BR mRNA的相对表达量高于CCK—AR;LPS作用2 h可明显诱导2种CCK受体mRNA表达上调,至12 h仍维持较高水平。用γ-32P—ATP标记的两种特异性探针分别对CCK—AR和CCK—BR mRNA RT—PCR扩增产物进行Southern分子杂交,结果均有特异性杂交带出现。结论:肺间质巨噬细胞有CCK—AR和CCK—BR mRNA表达,LPS可诱导2种CCK受体mRNA表达上调。
Objective: To investigate the expression of cholecystokinin (CCK) receptor mRNA and the effect of lipopolysaccharides(LPS) on its expression in rat pulmonary interstitial macrophages (PIMs). Methods: The PIMs isolated from rat lung tissues were purified by the collagenase digestion method combined with alveolar lavage and pulmonary vessel perfusion, and incubated with LPSQO mg/L) for 0. 5-12 h. The subtypes of CCK receptor and the effects of LPS on its expression were investigated by RT-PCR and Southern blot analysis in rat PIMs. Results: CCK-AR and CCK-BR mRNA were detected in rat PIMs by means of RT-PCR; their amplification products were approximately 1. 37 kb and 480 bp,respectively. The expression of CCK-BR mRNA was higher than that of CCK-AR mRNA,and the expression of CCK-AR and CCK-BR mRNA could be up-regulated obviously by LPS incubation for 2 to 12 h. r-32P-ATP 5'-end-labelled probes showed specific hybridization bands by Southern blot analysis of CCK-AR and CCK-BR mRNA products. Conclusion: There are CCK-AR and CCK-BR mRNA expressions in rat PIMs and the expressions can be up-regulated by LPS.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2003年第11期1208-1211,共4页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(30270529)
河北省自然科学基金(303452).
