Effects of human serum albumin complexed with free fatty acids on cell viability and insulin secretion in the hamster pancreatic β-cell line HIT-T15

被引:10
|
作者
Tuei, Vivian C. [1 ,2 ]
Ha, Ji-Sook [3 ]
Ha, Chung-Eun [1 ,2 ]
机构
[1] Univ Hawaii Manoa, John A Burns Sch Med, Dept Native Hawaiian Hlth, Honolulu, HI 96822 USA
[2] Univ Hawaii Manoa, Dept Mol Biosci & Bioengn, Coll Trop Agr & Human Resources, Honolulu, HI 96822 USA
[3] Univ Hawaii Manoa, John A Burns Sch Med, Dept Med Technol, Honolulu, HI 96822 USA
关键词
Human serum albumin; Free fatty acid; HIT-T15; cell; Insulin secretion; Cell viability; In vitro protein expression; Site-directed mutagenesis; ADIFAB; ENDOPLASMIC-RETICULUM STRESS; PALMITATE-INDUCED APOPTOSIS; FLUORESCENT-PROBE ADIFAB; CYTOPROTECTIVE ACTIONS; GLUCOSE-CONCENTRATION; CRYSTAL-STRUCTURE; CYCLOSPORINE-A; BINDING-SITES; ER STRESS; MECHANISMS;
D O I
10.1016/j.lfs.2011.02.022
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Aims: The effects of human serum albumin (HSA) complexed with various free fatty acids (FFAs) on g-cells have not been studied in detail. In this study, we examined the effects of HSA and its mutants on FFA-induced cell viability changes and insulin secretion from the hamster pancreatic insulinoma cell line, HIT-T15. Main methods: Cells were exposed to different FFAs in the presence of HSA or its mutants and/or bovine serum albumin (BSA) for 24 h. Cell viability, apoptosis, insulin secretion, and unbound FFA (FFAu) levels were determined. Key findings: In the presence of 0.1 mM HSA, palmitate and stearate induced significant cell death at 0.1 mM or higher, whereas myristate, palmitoleate, oleate, elaidate, linoleate, linoelaidate, and conjugated linoleate showed minimal changes on cell viability. Furthermore, oleate and linoleate were clearly cytoprotective against palmitate-induced cell death. The apoptosis inhibitors, cyclosporin A (csA) and the caspase inhibitor ZVAD-FMK, did not completely prevent FFA-induced cell death, although ZVAD-FMK blocked apoptosis with no differences in the presence of either HSA or BSA. In addition, insulin secretion from the cells was significantly reduced in the presence of HSA/oleate complexes. We also found differential effects of HSA mutants complexed with FFAs on cell viability. Significance: In summary, our results showed that saturated FFAs induced more cell death than unsaturated FFAs. Furthermore, modified HSA/FFA interactions caused by mutations of key amino acids involved in the binding of FFA to HSA resulted in changes in cell viability, suggesting a possible role of HSA polymorphism on FFA-induced changes in cellular functions. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码:810 / 818
页数:9
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