摘要
目的:观察肝素对脂多糖(lipopolysaccharide,LPS)刺激的人内皮细胞白细胞介素8(interleukin-8,IL-8)水平的影响,并探讨Toll样受体4(Toll-like receptor 4,TLR 4)在其中的可能影响。方法:用LPS(10mg/L)刺激人肺微血管内皮细胞诱导损伤,肝素治疗组提前15 min分别加入100 U/L及103U/L普通肝素,正常对照组加入等量磷酸盐缓冲液。分别在刺激2、6、12 h收集细胞上清,采用酶联免疫吸附法测定上清中IL-8的浓度。在刺激2、6、12 h收集细胞提取RNA,应用实时荧光定量聚合酶链反应检测各组细胞中IL-8、CD14及TLR4mRNA水平变化。结果:与正常对照组比较,LPS刺激组IL-8 mRNA水平增高,6 h达到高峰,其蛋白水平于12 h达到高峰。LPS刺激下TLR4 mRNA水平增高,6 h达到高峰,肝素降低其水平,差异有统计学意义(P<0.05)。未检测到CD14 mRNA的表达。结论:LPS刺激下人肺微血管内皮细胞IL-8表达增加。肝素可能通过调节TLR4降低IL-8的水平,从而发挥保护作用。
AIM：To determine the effect of unfractionated heparin（UFH） on lipopolysaccharide（LPS）-induced expression of interleukin-8（IL-8） and the role of Toll-like receptor 4（TLR4） signaling. METHODS：Human pulmonary microvascular endothelial cells（HPMECs） were either exposed to LPS alone（10 mg/L） or in combination with 100 U/L or 103 U/L UFH. UFH was added to the cells 15 min prior to stimulation with LPS. Those samples not receiving LPS or UFH received an equal volume of phosphate-buffered saline. The concentrations of IL-8 in the cell culture supernatants were detected by ELISA at 2, 6 and 12 h. The mRNA expression of IL-8, CD14 and TLR4 in HPMECs was detected by real-time fluorescence quantitative polymerase chain reaction at 2, 6 and 12 h. RESULTS：Compared with normal control group, the mRNA expression of IL-8 in LPS group was increased, and reached the peak at 6 h. The protein level of IL-8 reached the peak at 12 h. The mRNA expression of TLR4 in LPS group reached the peak at 6 h. They were down-regulated in UFH group. The mRNA expression of CD14 was not detected. CONCLUSION：The expression of IL-8 is obviously increased in LPS-treated HPMECs. UFH might exert its therapeutic effect through TLR4 signaling.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第9期1699-1701,共3页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81101411)
作者简介
通讯作者Tel：024—83282261；E—mail：xcma2972@sina．com
