周龍 王立林 李亞文 等
[摘要] 目的 探討丙泊酚對小鼠原代肝細胞PTEN表達的影響。 方法 采用兩步膠原酶灌注法分離小鼠原代肝細胞,均分為三組:對照組(C組)、溶劑對照組(D組)、丙泊酚組(P組)。首先用MTT法檢測不同培養(yǎng)濃度[(1~100)μg/mL]下丙泊酚對小鼠原代肝細胞活性的影響,接著選取10 μg/mL的丙泊酚(終濃度)作為培養(yǎng)條件,用MTT法檢測不同培養(yǎng)時間[(1~32)h]下小鼠原代肝細胞活力變化。P組選用10 μg/mL的丙泊酚(終濃度)處理小鼠原代肝細胞24 h。用Real-time PCR檢測各組PTEN mRNA表達,用Western blot檢測各組PTEN蛋白含量。 結(jié)果 在所有檢測項目上,C組與D組的差異均無統(tǒng)計學(xué)意義(P>0.05)。與C組相比,丙泊酚濃度為1 μg/mL、5 μg/mL、10 μg/mL、25 μg/mL時小鼠原代肝細胞活性的差異無統(tǒng)計學(xué)意義(P>0.05),丙泊酚濃度為100 μg/mL時小鼠原代肝細胞活性顯著降低(P<0.01)。不同培養(yǎng)時間的各組與C組相比,小鼠原代肝細胞活性的差異無統(tǒng)計學(xué)意義(P>0.05)。P組PTEN蛋白含量顯著高于D組(P<0.01),P組PTEN mRNA水平高于D組(P<0.05)。 結(jié)論 丙泊酚可引起小鼠原代肝細胞PTEN表達增強。
[關(guān)鍵詞] 二異丙酚;小鼠原代肝細胞;PTEN磷酸水解酶;胰島素抵抗
[中圖分類號] R575.5? ? ? ? ? [文獻標(biāo)識碼] A? ? ? ? ? [文章編號] 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.