脂联素影响体外培养肝细胞乙醛氧化酶1表达的研究

Investigation on the influence of adiponectin on aldehyde oxidase-1 expression of hepatic cell lines in vitro

目的研究脂联素(APN)对饱和脂肪酸(SFA)孵育HepG2细胞及HL-7701肝细胞乙醛氧化酶1(AOX1)分泌的影响,探讨APN对非酒精性脂肪肝脏病变(NAFLD)的治疗效果及机制。方法用100μmol/LSFA孵育细胞得到脂肪性HepG2细胞及HL-7701细胞,同时设立细胞对照组(未经100μmol/L SFA孵育)。然后采用不同浓度APN(0.5、1.0、1.5、2.5μg/mL)孵育脂肪性HepG2细胞及HL-7701细胞,将未经APN孵育的细胞作为对照。运用逆转录聚合酶链反应(RT-PCR)检测2种细胞AOX1基因表达,运用免疫印迹法检测2种细胞AOX1蛋白表达。结果与未经100μmol/L SFA孵育的对照组比较,经100μmol/L SFA孵育过的HepG2细胞及HL-7701细胞的AOX1基因及蛋白表达均明显增加(P<0.05),且HepG2细胞组明显高于HL-7701细胞组(P<0.05)。加入APN孵育细胞后,2种细胞AOX1基因及蛋白表达均明显低于未经APN孵育的对照组(P<0.05);并且随APN浓度升高,降幅增加。当APN浓度达到1.5μg/mL时,细胞AOX1表达的降低不再随APN浓度的增加而发生明显变化。在APN浓度相同的条件下,HepG2细胞组中AOX1基因及蛋白表达降低的幅度明显高于HL-7701细胞组。结论 APN能有效降低脂肪化肝脏细胞AOX1基因及蛋白表达,能很好的保护肝脏细胞,避免肝脏细胞发生NAFLD。

Objective To investigate the influence of adiponectin（APN） on aldehyde oxidase-1（AOX1） expression of HepG2 hepatoma cell lines and HL-7701 hepatic cell lines incubated by saturated fatty acid（SFA）,and discuss the curative effect and possible mechanism of APN to protect non-alcohol fatty liver disease（NAFLD）.Methods Fatty HepG2 hepatoma cell lines and HL-7701 hepatic cell lines were incubated by 100 μmol/L SFA,and cells without being incubated by 100 μmol/L SFA were as controls.The fatty HepG2 hepatoma cell lines and HL-7701 hepatic cell lines were performed the treatment of APN（0.5,1.0,1.5 and 2.5 μg/mL）,and cells without APN treatment were used as control group.Reverse transcription-polymerase chain reaction（RT-PCR） and Western-blot were utilized to detect the expressions of AOX1 gene and AOX1 protein.Results The AOX1 gene and AOX1 protein expressions of cells incubated by 100 μmol/L SFA were significantly higher than those of controls without being incubated by 100 μmol/L SFA（P0.05）,and the expression of HepG2 cells was significantly higher than that of HL-7701 cells（P0.05）.The AOX1 gene and AOX1 protein expressions of cells treated by APN were reduced significantly comparing to those without APN treatment（P0.05）,which indicated that APN could decrease the expression effectively.However,the AOX1 expression of cells had no change,when the APN concentration reached 1.5 μg/mL.The AOX1 gene and AOX1 protein expressions of HepG2 cells decreased significantly higher than those of HL-7701 cells at the same APN concentration.Conclusions APN could reduce the expression of cells incubated by saturated fatty acid effectively,therefore it could protect hepatic cells from NAFLD.