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目的观察甲醛对肝癌细胞株HepG2细胞内外甘油三酯(TG)含量及细胞内极低密度脂蛋白(VLDL)分泌外排相关基因表达水平的影响。方法以0、0.004、0.02、0.1 mmol/L甲醛分别处理人肝癌HepG2细胞24、48 h,采用甘油磷酸氧化酶法(GPO-POD)测定细胞内外TG含量,采用QRT-PCR法检测细胞内载脂蛋白B100(apo B100)、微粒体TG转运蛋白(MTTP)、载脂蛋白E(apo E)、TG水解酶(CES3)、二酰基甘油酰转移酶1(DGAT1)、包被蛋白Ⅱ(COPⅡ)、Sar1b、低密度脂蛋白受体(LDLR)基因的表达水平。结果与阴性对照组相比,0.004、0.1 mmol/L甲醛染毒48 h后HepG2细胞内TG含量减少,染毒24 h后上清中TG含量增加;染毒24 h后0.004、0.02、0.1 mmol/L甲醛组或染毒48 h后0.004 mmol/L甲醛组细胞内apo B100基因表达水平升高;染毒24 h后0.004 mmol/L甲醛组或染毒48 h后0.004、0.02、0.1 mmol/L甲醛组细胞内MTTP基因表达水平升高;染毒24 h后0.1 mmol/L甲醛组细胞内LDLR基因表达水平降低,染毒48 h后0.02、0.1 mmol/L甲醛组细胞内LDLR基因表达水平升高;上述差异均有统计学意义(P<0.05)。结论甲醛可能通过增加肝细胞内apo B100和MTTP表达来促进VLDL的分泌外排。
Objective To investigate the effect of formaldehyde on the content of triglyceride (TG) and the level of extracellular secretion of extracellular very low density lipoprotein (VLDL) in hepatocellular carcinoma cell line HepG2. Methods HepG2 cells were treated with 0, 0.004, 0.02 and 0.1 mmol / L formalin for 24 and 48 h respectively. The contents of intracellular and extracellular TG were measured by glycerophosphate oxidase method (GPO-POD) (Apo B100), microsomal TG transporter (MTTP), apolipoprotein E (apo E), TG hydrolase (CES3), diacylglycerol acyltransferase 1 (DGAT1), coat Ⅱ ), Sar1b, low density lipoprotein receptor (LDLR) gene expression levels. Results Compared with the negative control group, the content of TG in HepG2 cells decreased after 0.004 and 0.1 mmol / L formaldehyde exposure for 48 h, and the content of TG in the supernatant increased 24 h after exposure. After 24 h exposure, 0.004, 0.02 and 0.1 mmol After treatment with 0.004 mmol / L formaldehyde or 0.004 mmol / L formaldehyde for 24 h, the expression levels of apo B100 in 0.004 mmol / L formaldehyde group increased significantly L formaldehyde group increased intracellular MTTP gene expression; 24 hours after exposure to 0.1 mmol / L formaldehyde group decreased the level of LDLR gene expression, within 48 hours after exposure to 0.02,0.1 mmol / L formaldehyde group intracellular LDLR gene expression levels Increased; the above differences were statistically significant (P <0.05). Conclusion Formaldehyde may promote secretion of VLDL by increasing the expression of apo B100 and MTTP in hepatocytes.