丙泊酚对小鼠原代肝细胞PTEN表达的影响

周龙 王立林 李亚文 等

[摘要] 目的 探讨丙泊酚对小鼠原代肝细胞PTEN表达的影响。 方法 采用两步胶原酶灌注法分离小鼠原代肝细胞，均分为三组：对照组（C组）、溶剂对照组（D组）、丙泊酚组（P组）。首先用MTT法检测不同培养浓度[（1～100）μg/mL]下丙泊酚对小鼠原代肝细胞活性的影响，接着选取10 μg/mL的丙泊酚（终浓度）作为培养条件，用MTT法检测不同培养时间[（1～32）h]下小鼠原代肝细胞活力变化。P组选用10 μg/mL的丙泊酚（终浓度）处理小鼠原代肝细胞24 h。用Real-time PCR检测各组PTEN mRNA表达，用Western blot检測各组PTEN蛋白含量。 结果 在所有检测项目上，C组与D组的差异均无统计学意义（P>0.05）。与C组相比，丙泊酚浓度为1 μg/mL、5 μg/mL、10 μg/mL、25 μg/mL时小鼠原代肝细胞活性的差异无统计学意义（P>0.05），丙泊酚浓度为100 μg/mL时小鼠原代肝细胞活性显著降低（P<0.01）。不同培养时间的各组与C组相比，小鼠原代肝细胞活性的差异无统计学意义（P>0.05）。P组PTEN蛋白含量显著高于D组（P<0.01），P组PTEN mRNA水平高于D组（P<0.05）。 结论 丙泊酚可引起小鼠原代肝细胞PTEN表达增强。

[关键词] 二异丙酚;小鼠原代肝细胞;PTEN磷酸水解酶;胰岛素抵抗

[中图分类号] R575.5          [文献标识码] A          [文章编号] 1673-9701（2019）03-0030-04

[Abstract] Objective To explore the effect of propofol on the expression of PTEN in primary hepatocytes of mice. Methods Mouse primary hepatocytes were isolated by two-step collagenase perfusion and divided into three groups： control group （group C）， solvent control group （group D）， and propofol group （group P）. Firstly， MTT assay was used to detect the effect of propofol on the activity of primary hepatocytes in mice at different culture concentrations [（1-100） μg/mL）. Then 10 μg/mL propofol （final concentration） was selected as the culture condition. MTT assay was used to detect the changes of primary hepatocyte viability in mice under different culture time [（1-32）h]. In group P， 10 μg/mL propofol （final concentration） was used to treat mouse primary hepatocytes for 24 h. The expression of PTEN mRNA in each group was detected by Real-time PCR， and the content of PTEN protein in each group was detected by Western blot. Results There was no significant difference between group C and group D in all the test items（P>0.05）. Compared with group C， there was no significant difference in primary hepatocyte activity when propofol concentrations were 1 μg/mL， 5 μg/mL， 10 μg/mL， and 25 μg/mL（P>0.05）. When the concentration of propofol was 100 μg/mL，the activity of primary hepatocytes was significantly decreased（P<0.01）. There was no significant difference in primary hepatocyte activity between groups with different culture time and group C（P>0.05）. The PTEN protein content in group P was significantly higher than that in group D（P<0.01）. The PTEN mRNA level in group P was higher than that in group D（P<0.05）. Conclusion Propofol can cause an increase in PTEN expression in primary hepatocytes of mice.