本研究旨在研究蛋氨酸(Met)对HepG2细胞和鸭原代肝细胞生长及脂质代谢的影响。设置2个体外肝细胞试验,分别以HepG2细胞和鸭原代肝细胞为研究对象,研究不同浓度Met培养基对HepG2细胞和鸭原代肝细胞的存活率、脂质沉积及其相关基因和蛋白表达的影响。结果表明:1)培养基中Met浓度为0 μmol/L时HepG2细胞存活率显著低于其他Met浓度(P<0.05),培养基中Met浓度为0、6.25、12.5和25 μmol/L时鸭原代肝细胞存活率显著低于显著100和200 μmol/L时(P<0.05),随着培养基Met浓度增加,25 μmol/L Met时HepG2细胞存活率达到平台,100 μmol/L Met时鸭原代肝细胞存活率达到平台。2)与200 μmol/L Met组相比,0、25 μmol/L Met组脂滴数量明显增加,单位细胞数目油红O的吸光度显著增加(P<0.05),且0、25 μmol/L组之间无显著差异(P>0.05)。3)与200 μmol/L Met组相比,0、25 μmol/L Met组HepG2细胞中链酰基辅酶A脱氢酶(ACADM)、二氢硫辛酸脱氢酶(DLD)和泛醌还原型酰胺腺嘌呤二核苷酸脱氢酶Fe-S蛋白1(NDUFS1)基因表达量显著下调(P<0.05),且0 μmol/L Met组NDUFS1蛋白表达量显著下调(P<0.05)。4)与200 μmol/L Met组相比,0、25、100 μmol/L Met组鸭原代肝细胞中苹果酸脱氢酶(MDH)1、MDH2、DLD、3-羟基-3-甲基戊二酰辅酶A合成酶2(HMGCS2)、电子转移黄素蛋白α亚基(ETFA)、电子传递黄素蛋白脱氢酶(ETFED)和NDUFS1基因表达量显著下调(P<0.05),且0、25 μmol/L Met组NDUFS1蛋白表达量显著下调(P<0.05)。综上所述,Met缺乏可导致HepG2细胞和鸭原代肝细胞生长受阻和脂质沉积增加,主要由于肝细胞中脂肪酸β-氧化、三羧酸循环、呼吸链电子传递和酮体生成等过程受阻,导致肝细胞能量供应不足。
吴永保
,
唐静
,
闻治国
,
曹俊婷
,
张博
,
邢光楠
,
谢明
,
胡海峰
,
崔德福
,
侯水生
. 蛋氨酸对HepG2细胞和鸭原代肝细胞生长和脂质代谢的影响[J]. 动物营养学报, 2022
, 34(8)
: 5364
-5373
.
DOI: 10.3969/j.issn.1006-267x.2022.08.056
This experiment was conducted to investigate the effects of methionine (Met) on growth and lipid metabolism in HepG2 cells and duck primary hepatocytes. There were two in vitro hepatocyte experiments, including HepG2 cells and duck primary hepatocytes, to study the effects of different Met concentration in medium on the cell viability, lipid deposition and related gene and protein expression of HepG2 cells and duck primary hepatocytes. The results showed as follows: 1) the viability of HepG2 cells when the medium Met concentration 0 μmol/L was significantly lower than other Met concentration (P<0.05), the viability of duck primary hepatocytes when the medium Met concentration 0, 6.25, 12.5 and 25 μmol/L was significantly lower than 100 and 200 μmol/L (P<0.05), with the increased of Met concentration in medium, the plateaus were obtained at 25 and 100 μmol/L Met for HepG2 cells and duck primary hepatocytes, respectively. 2) Compared with 200 μmol/L Met group, the lipid droplet number was increased, the absorbance of oil red O per cell of 0 and 25 μmol/L Met groups was significantly increased (P<0.05), and no difference was observed between 0 and 25 μmol/L Met groups (P>0.05). 3) Compared with 200 μmol/L Met group, the gene expression levels of medium chain acyl-CoA dehydrogenase (ACADM), dihydrolipoyl dehydrogenase (DLD), reduced amide adenine dinucleotide-dehydrogenase ubiquinone Fe-S protein 1 (NDUFS1) in HepG2 cells of 0 and 25 μmol/L Met groups were significantly down regulated (P<0.05), and the NDUFS1 protein expression level of 0 μmol/L Met group was significantly down regulated (P<0.05). 4) Compared with 200 μmol/L Met group, the gene expression levels of malic dehydrogenase (MDH)1, MDH2, DLD, 3-hydroxy-3-methylglutaryl-coA synthase 2 (HMGCS2), electron transfer flavin protein alpha subunit (ETFA), electron transport flavin protein dehydrogenase (ETFED) and NDUFS1 in duck primary hepatocytes of 0, 25 and 100 μmol/L Met groups were significantly down regulated (P<0.05), and the NDUFS1 protein expression level of 0 and 25 μmol/L Met groups was significantly down regulated (P<0.05). It is concluded that Met deficiency can lead to growth inhibition and lipid deposition increasing in HepG2 cells and duck primary hepatocytes, which mainly due to the processes of fatty acid β-oxidation, tricarboxylic acid cycle, respiratory chain electron transport and ketoplasia are depressed, resulting in insufficient energy supply in hepatocytes.
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