Determining the temporal, dose, and composition effects of nutritional substrates in an in vitro model of intrahepatocellular triglyceride accumulation.
Determining the temporal, dose, and composition effects of nutritional substrates in an in vitro model of intrahepatocellular triglyceride accumulation.
复制标题
DOI：
10.14814/phy2.15463
复制
发表时间：
2022-10
影响因子：
2.5
通讯作者：
中科院分区：
文献类型：
--
作者：
肝内甘油三酯的病理性积累是非酒精性脂肪性肝病（NAFLD）早期阶段的基础，并可进展为肝纤维化、肝硬化和肝癌。对人类的研究表明，与不饱和脂肪酸（FA）相比，富含饱和脂肪酸的饮食对肝脏脂肪积累和代谢更有害。然而，分歧的原因仍不清楚，需要生理相关的细胞模型。因此，本研究的目的是调查修改媒体组成，浓度和治疗频率的糖，脂肪酸和胰岛素对肝细胞内甘油三酯含量和细胞内葡萄糖，脂肪酸和昼夜节律功能的影响。Huh 7细胞用2%人血清和富含不饱和（OPLA）或饱和（POLA）FA的糖和FA的组合（低脂低糖[LFLS]、高脂低糖[HFLS]或高脂高糖[HFHS]）处理2、4或7天，每天或交替处理方案。使用稳定同位素示踪剂研究不同给药方案下肝细胞代谢的基础和/或胰岛素反应性变化。定量细胞活力、培养基生物化学、细胞内代谢和昼夜节律生物学。培养基的FA组成（OPLA与POLA）不影响肝细胞中的细胞活力或细胞内甘油三酯含量。相比之下，与OPLA处理的细胞相比，POLA处理的细胞具有较低的FA氧化和培养基乙酸盐，并且具有较高的FA浓度，显示出较低的细胞内糖原含量和减少的胰岛素刺激糖原生成。HFHS的加入也对昼夜节律振荡和基因表达产生了深远的影响。每天用HFHS处理细胞至少4天，导致细胞模型显示出在人类中观察到的早期NAFLD的特征。更长时间（≥7天）的重复治疗可能为研究NAFLD更严重阶段的脂质和葡萄糖代谢提供机会。
Pathological accumulation of intrahepatic triglyceride underpins the early stages of nonalcoholic fatty liver disease (NAFLD) and can progress to fibrosis, cirrhosis, and cancer of the liver. Studies in humans suggest that consumption of a diet enriched in saturated compared to unsaturated fatty acids (FAs), is more detrimental to liver fat accumulation and metabolism. However, the reasons for the divergence remain unclear and physiologically‐relevant cellular models are required. Therefore, the aims of this study were to investigate the effect of modifying media composition, concentration, and treatment frequency of sugars, FAs and insulin on intrahepatocellular triglyceride content and intracellular glucose, FA and circadian function. Huh7 cells were treated with 2% human serum and a combination of sugars and FAs (low fat low sugar [LFLS], high fat low sugar [HFLS], or high fat high sugar [HFHS]) enriched in either unsaturated (OPLA) or saturated (POLA) FAs for 2, 4, or 7 days with a daily or alternating treatment regime. Stable isotope tracers were utilized to investigate basal and/or insulin‐responsive changes in hepatocyte metabolism in response to different treatment regimes. Cell viability, media biochemistry, intracellular metabolism, and circadian biology were quantified. The FA composition of the media (OPLA vs. POLA) did not influence cell viability or intracellular triglyceride content in hepatocytes. In contrast, POLA‐treated cells had lower FA oxidation and media acetate, and with higher FA concentrations, displayed lower intracellular glycogen content and diminished insulin stimulation of glycogenesis, compared to OPLA‐treated cells. The addition of HFHS also had profound effects on circadian oscillation and gene expression. Cells treated daily with HFHS for at least 4 days resulted in a cellular model displaying characteristics of early stage NAFLD seen in humans. Repeated treatment for longer durations (≥7 days) may provide opportunities to investigate lipid and glucose metabolism in more severe stages of NAFLD.
登录
查看更多内容
影响因子：
7.1
作者：
Bjermo, Helena;Iggman, David;Riserus, Ulf
通讯作者：
Riserus, Ulf
影响因子：
29.4
作者：
Lambert JE;Ramos-Roman MA;Browning JD;Parks EJ
通讯作者：
Parks EJ
影响因子：
29.4
作者：
Cotter, Thomas G.;Rinella, Mary
通讯作者：
Rinella, Mary
DOI：
10.1016/s0006-291x(02)02140-x
发表时间：
2002-09-13
影响因子：
3.1
作者：
Chirieac, DV;Cianci, J;Sparks, CE
通讯作者：
Sparks, CE
影响因子：
5
作者：
Eynaudi A;Díaz-Castro F;Bórquez JC;Bravo-Sagua R;Parra V;Troncoso R
通讯作者：
Troncoso R