Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells.
Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells.
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啮齿动物与人β细胞中长链饱和脂肪酸棕榈酸酯的差异路由和处置。
DOI：
10.1038/s41387-022-00199-y
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发表时间：
2022-04-20
影响因子：
6.1
通讯作者：
Morgan, Noel G.
中科院分区：
文献类型：
--
作者：
Thomas, Patricia;Arden, Catherine;Corcoran, Jenna;Hacker, Christian;Welters, Hannah J.;Morgan, Noel G.
啮齿动物和人类的β细胞对长链饱和脂肪酸(LC-SFA)的“脂毒”作用有不同的敏感性，但造成这种影响的因素尚不清楚。在这里，我们研究了LC-SFA棕榈酸酯在人和啮齿动物β细胞中的细胞内分布，并提供了对调节β细胞脂毒性的因素的新见解。用共聚焦荧光和电子显微镜研究了LC-SFA棕榈酸酯在啮齿类动物(INS-1E和INS-1823/13细胞)和人(EnDoc-βH1)β细胞中的亚细胞分布。蛋白表达采用免疫印迹法检测，细胞存活率通过活性染料染色检测。INS-1细胞暴露于棕榈酸酯24 h会导致细胞活力丧失，而Endo-βH1细胞即使在72 h的高浓度棕榈酸酯(1 mM)处理后仍能存活。使用荧光棕榈酸酯类似物BODIPY FL C16揭示了LC-SFA在INS-1细胞中高尔基体的早期定位，这与细胞内膜的扩张有关，在EM下可见。尽管如此，依赖于PERK的内质网应激通路在这些条件下并未被激活。相比之下，BODIPY FL C16没有在EnDoc-βH1细胞的高尔基体中积累，而是与脂滴相关蛋白PLIN2共同定位，表明优先进入脂滴。当用棕榈酸酯和油酸盐联合处理INS-1细胞时，棕榈酸酯的毒性作用减弱，BODIPFL C16主要与PLIN2结合，但不与高尔基标记结合。在啮齿动物β细胞中，棕榈酸酯在早期时间点积累在高尔基体中，而在EnDoc-βH1细胞中，棕榈酸酯优先进入脂滴。这可能解释了啮齿动物和人类β细胞对“脂毒性”的不同敏感性，因为导致棕榈酸酯进入脂滴的动作与维持这两种细胞的细胞活力有关。
Rodent and human β-cells are differentially susceptible to the “lipotoxic” effects of long-chain saturated fatty acids (LC-SFA) but the factors accounting for this are unclear. Here, we have studied the intracellular disposition of the LC-SFA palmitate in human vs rodent β–cells and present data that reveal new insights into the factors regulating β-cell lipotoxicity. The subcellular distribution of the LC-SFA palmitate was studied in rodent (INS-1E and INS-1 823/13 cells) and human (EndoC-βH1) β-cells using confocal fluorescence and electron microscopy (EM). Protein expression was assessed by Western blotting and cell viability, by vital dye staining. Exposure of INS-1 cells to palmitate for 24 h led to loss of viability, whereas EndoC-βH1 cells remained viable even after 72 h of treatment with a high concentration (1 mM) of palmitate. Use of the fluorescent palmitate analogue BODIPY FL C16 revealed an early localisation of the LC-SFA to the Golgi apparatus in INS-1 cells and this correlated with distention of intracellular membranes, visualised under the EM. Despite this, the PERK-dependent ER stress pathway was not activated under these conditions. By contrast, BODIPY FL C16 did not accumulate in the Golgi apparatus in EndoC-βH1 cells but, rather, co-localised with the lipid droplet-associated protein, PLIN2, suggesting preferential routing into lipid droplets. When INS-1 cells were treated with a combination of palmitate plus oleate, the toxic effects of palmitate were attenuated and BODIPY FL C16 localised primarily with PLIN2 but not with a Golgi marker. In rodent β-cells, palmitate accumulates in the Golgi apparatus at early time points whereas, in EndoC- βH1 cells, it is routed preferentially into lipid droplets. This may account for the differential sensitivity of rodent vs human β-cells to “lipotoxicity” since manoeuvres leading to the incorporation of palmitate into lipid droplets is associated with the maintenance of cell viability in both cell types.
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影响因子：
6.5
作者：
Huber, Andrew H.;Kleinfeld, Alan M.
通讯作者：
Kleinfeld, Alan M.
影响因子：
4.5
作者：
Plötz T;Hartmann M;Lenzen S;Elsner M
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Elsner M
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4
作者：
Diakogiannaki, Eleftheria;Welters, Hannah J.;Morgan, Noel G.
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Morgan, Noel G.
影响因子：
8.2
作者：
Marchetti, P.;Bugliani, M.;Cnop, M.
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Cnop, M.
影响因子：
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Carta G;Murru E;Banni S;Manca C
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