丙泊酚对缺氧肝细胞复氧后核因子κB活性和细胞凋亡的影响

Influence of propofol on activation of nuclear factor-κB and cell apoptosis induced by hypoxia-reoxygenation in hepatocytes

目的观察临床相关浓度丙泊酚对缺氧肝细胞复氧后核因子κB诱导激活和细胞凋亡的影响。方法采用携带报告基因的6κB-TK-Luc质粒转染离体培养的LO2肝细胞，随机数字表法分为3组：正常对照组（CO组）、缺氧复氧组（HR组）、二硫代氨基甲酸吡咯烷（PDTC，50M）阻断组（PD组），每组按加入丙泊酚浓度不同分为4个亚组（终末浓度为0、125、25、50M）。除CO组正常培养外，其余各组均给予缺氧4h后复氧8h处理。分别测定KB序列依赖性报告基因虫荧光素酶（Luc）的转录表达水平和肝细胞凋亡率。结果氧肝细胞复氧后Luc表达水平和细胞凋亡率均较CO组明显升高（P〈0．01）。不同浓度的丙泊酚明显降低缺氧肝细胞复氧后的Luc表达水平（P〈0.05），且呈浓度依赖性，但其作用可被联合应用的PDTC所消除。较高浓度（25M和50M）的丙泊酚可有效抑制缺氧复氧处理后的肝细胞凋亡率（P〈0.05），而联合应用PDTC后，50M丙泊酚亚组仍显示有明显的抑制作用（P〈0.05）。结论较高浓度的丙泊酚（≥25M）可有效降低缺氧复氧后的肝细胞凋亡，其作用部分与抑制核因子κB的诱导激活有关。

Objective To investigate the Influence of propofol on activation of nuclear factor- κB and cell apoptosis induced by hypoxia-reoxygenation in hepatocytes. Methods Plasmid 6 κB-TK-Luc containing NF-κB-dependent reporter gene luciferase was transfected into cultured LO2 liver cells. The cultured hepatocytes were randomly assigned into 3 groups. group CO receiving normoxia as control;group HR receiving 4h hypoxia followed by 8h reoxygenation;group PD receiving hypoxia-reoxygenation and 50 M pyrrolidine dithiocarbamate （PDTC）. Each group was subdivided into 4 subgroups, separately treated with 0, 12.5, 25, 50 M of propofol. The expression of reporter gene luciferase and cell apoptosis induced by hypoxia-reoxygenation in hepatocytes was measured. Results Anoxia-reoxygenation caused dramatic increase in the expression of reporter gene luciferase and the rate of apoptotic cells as compared with that of control group （P〈0.01）. Propofol concentration-dependently inhibited the inducible expression of reporter gene luciferaser after hypoxia-reoxygenation in hepatocytes, but the effects were abolished by the combined apply of PDTC. Relatively high concentrations （25 M and 50 M） of propofol lowered the rate of apoptotic cells induced by hypoxia-reoxygenation, and even with the blocker of PDTC, the anti-apoptosis effect was still existed in the subgroup of 50M propofol. Conclusion Propofol can decrease hepatocyte apoptosis following anoxia and reoxygenation injury,which is associated with the inhibition on the inducible activation of NF-κB.