Fish oil-derived lipid emulsion induces RIP1-dependent and caspase 8-licensed necroptosis in IEC-6 cells through overproduction of reactive oxygen species.
Fish oil-derived lipid emulsion induces RIP1-dependent and caspase 8-licensed necroptosis in IEC-6 cells through overproduction of reactive oxygen species.
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鱼油衍生的脂肪乳剂通过活性氧的过量产生，诱导 IEC-6 细胞中 RIP1 依赖性和 caspase 8 许可的坏死性凋亡
DOI：
10.1186/s12944-018-0786-5
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发表时间：
2018-06-23
影响因子：
4.5
通讯作者：
Cai W
中科院分区：
文献类型：
--
作者：
Yan JK;Yan WH;Cai W
背景肠外营养支持过程中静脉注射脂肪乳剂（LE）可导致肠上皮细胞过度死亡。然而，作为新一代的LE，鱼油衍生的脂肪乳剂（FOLE）对肠上皮细胞死亡的影响仍然是难以捉摸的。方法肠上皮细胞（IEC-6细胞系）用FOLE（0.25-1%）处理24小时。CCK-8测定细胞存活率，并通过延时活细胞成像监测形态学变化。用Westernblot法检测受体相互作用蛋白1/3（RIP 1/3）和半胱氨酸天冬氨酸蛋白酶8（caspase 8）的表达，（其特征在于RIP 1/3复合物的组装沿着半胱天冬酶8的解离）通过免疫沉淀进行检测。另外，细胞内活性氧（ROS）的产生通过使用具有氧化敏感探针的ROS检测试剂盒（DCFH-DA）来检测。结果FOLE处理后4-8 h内呈剂量依赖性诱导细胞程序性坏死（necroptosis），而非凋亡。在FOLE处理的细胞中观察到RIP 1/3复合物的组装沿着caspase 8从RIP 1的解离。此外，FOLE诱导的细胞死亡通过抑制RIP 1而显著减轻，并且通过抑制半胱天冬酶8而进一步加重。此外，在细胞死亡之前，在FOLE处理的细胞中细胞内ROS的积累显著增加（与对照相比增加约5倍，p< 0.001），这可以通过抑制RIP 1来减弱（与FOLE相比减少约35%，p< 0.05）。我们的研究结果可能会提供新的见解，在PN支持的临床应用FOLE。
BackgroundExcessive cell death of enterocytes has been demonstrated to be partially associated with the intravenously-administrated lipid emulsions (LEs) during parenteral nutrition (PN) support. However, as a new generation of LE, the effect of fish oil-derived lipid emulsion (FOLE) on the death of enterocytes remains elusive.MethodsIntestinal epithelial cells (IEC-6 cell line) were treated with FOLE (0.25–1%) for 24 h. Cell survival was measured by CCK-8 assay, and morphological changes were monitored by time-lapse live cell imaging. The expression of receptor-interacting protein 1/3 (RIP1/3) and caspase 8 was assessed by westernblot, and the formation of necrosome (characterized by the assembly of RIP1/3 complex along with the dissociation of caspase 8) was examined by immunoprecipitation.Additionally, the production of intracellular reactive oxygen species (ROS) was detected by using a ROS detection kit with an oxidation-sensitive probe (DCFH-DA).ResultsFOLE dose-dependently induced non-apoptotic, but programmed necroctic cell death (necroptosis) within 4–8 h after treatment. The assembly of RIP1/3 complex along with the dissociation of caspase 8 from RIP1 was observed in FOLE-treated cells. Moreover, FOLE-induced cell death was significantly alleviated by inhibiting RIP1, and was further aggravated by inhibiting caspase 8. In addition, prior to cell death the accumulation of intracellular ROS was significantly increased in FOLE-treated cells (increased by approximately 5-fold versus control,p< 0.001), which could be attenuated by inhibiting RIP1 (decreased by approximately 35% versus FOLE,p< 0.05).ConclusionsFOLE induces RIP1-dependent and caspase 8-licensed necroptosis through overproduction of ROS in vitro. Our findings may provide novel insights into the clinical applications of FOLE during PN support.
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影响因子：
5
作者：
Guzy, Claudia;Schirbel, Anja;Sturm, Andreas
通讯作者：
Sturm, Andreas
DOI：
10.1056/nejmoa0907929
发表时间：
2010-05-06
期刊：
The New England journal of medicine
影响因子：
--
作者：
Sanyal AJ;Chalasani N;Kowdley KV;McCullough A;Diehl AM;Bass NM;Neuschwander-Tetri BA;Lavine JE;Tonascia J;Unalp A;Van Natta M;Clark J;Brunt EM;Kleiner DE;Hoofnagle JH;Robuck PR;NASH CRN
通讯作者：
NASH CRN
影响因子：
5.9
作者：
Raman M;Almutairdi A;Mulesa L;Alberda C;Beattie C;Gramlich L
通讯作者：
Gramlich L
影响因子：
64.8
作者：
Oberst, Andrew;Dillon, Christopher P.;Weinlich, Ricardo;McCormick, Laura L.;Fitzgerald, Patrick;Pop, Cristina;Hakem, Razq;Salvesen, Guy S.;Green, Douglas R.
通讯作者：
Green, Douglas R.
影响因子：
3.7
作者：
Harris JK;El Kasmi KC;Anderson AL;Devereaux MW;Fillon SA;Robertson CE;Wagner BD;Stevens MJ;Pace NR;Sokol RJ
通讯作者：
Sokol RJ