摘要
目的在脂多糖（1ipopolysaccharide，LPS）诱导的人肺微血管内皮细胞（human pulmonary microvascular endothelial cells，，HPMVCs）损伤模型中，观察异丙酚预处理对血管紧张素转化酶（ angiotensin-converting enzyme，ACE）的影响，探讨异丙酚肺保护作用的可能机制。方法以体外培养的HPMVCs 3～5代作为实验对象，将细胞随机分为4组（n=8）：对照组（C组）、异丙酚组（P组）、LPS组（L组）和异丙酚预处理组（P＋L组）。药物终浓度：LPS5μg／mL，异丙酚50μmol／L。按上述分组，加入LPS前1h加入异丙酚。于37℃、5％CO2培养箱中进行培养。分别培养12、24、48、72h后采用CCK-8（Cell Counting Kiv-8）法测细胞活力。细胞孵育12h后，采用改良分光光度法检测各组培养液和细胞中的ACE活性，real-timePCR检测ACE、TNF-α、IL-1β和MCP-1mRNA表达水平，同时ELISA测定细胞培养液中TNF-α、IL-1β和MCP-1的蛋白含量。结果（1）各组细胞在72h内的细胞活力无显著差异（P〉0．05）；（2）与C组相比，P组各检测指标均无显著差异（P〉0．05）；（3）与c组相比，L组TNF-α、IL-1β和MCP-1的蛋白含量及其基因表达水平均显著升高，分泌型ACE活性增加，细胞ACE活性降低，ACEmRNA表达下调（P〈0．05）；（4）与L组相比，P＋L组分泌的TNF-α、IL-1β和MCP-1及其mRNA表达显著降低，细胞ACE活性增加，ACEmRNA表达上调（P〈0．05），分泌型ACE活性不变（P〉0．05）。结论异丙酚在转录水平调节HPMVCs中ACE的表达，可能是异丙酚保护HPMVCs的作用机制之一。
Objective To investigate the effects of propofol pretreatment on the angiotensin- converting enzyme （ACE） of human pulmonary microvascular endothelial cells （HPMVCs） induced by lipopolysaccharide （LPS）. Methods HPMVCs were randomly assigned to one of the following 4 groups （n = 8 each） ：control group,propofol （50μool/L） ,LPS （5 μg/mL） and propofol （50 μmol/L） combined with LPS. Propofol was administered 1 h before LPS. After being incubated for 12,24,48 and 72 h, cell viability was determined by Cell Counting Kit-8 （CCK-8） assay. After the incubationof 12 h, ACE activity in culture medium as well as in cell lysate were determined by an improved spectrophotometric method, real-time PCR was applied for detecting the mRNA expression of ACE, TNF-α, IL-1β and MCP-1 ,and the protein concentration of TNF-α, IL-1β and MCP-1 in medium was detected with ELISA kits. Results Survival of HPMVCs was not influenced by drugs treated for 12, 24,48 and 72 h in this experiment （P〉0.05）. Propofol alone had no significant effect on HPMVCs in terms of ACE activity and the expression of ACE, as well as the expression of TNF-α, IL-1βand MCP- 1 in both protein and gene level （P〉0.05）. LPS significantly increased ACE activity in the medium and decreased both cellular ACE activity and ACE mRNA, with an increased expression of TNF-α, IL-1β and MCP-1 （P〈0. 05）. Pretreatment of propofol significantly attenuated LPS-induced effects on HPMVCs （P 〈0. 05）. Conclusions Propofol may take part in regulating ACE expression of HPMVCs challenged by LPS in vitro at the transcriptional level.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2013年第6期668-672,共5页
Fudan University Journal of Medical Sciences
基金
上海市科委科研计划项目(o841196030)~~
