The Caspase Pathway of Linoelaidic Acid (9t, 12t-C18:2)-Induced Apoptosis in Human Umbilical Vein Endothelial Cells
The Caspase Pathway of Linoelaidic Acid (9t, 12t-C18:2)-Induced Apoptosis in Human Umbilical Vein Endothelial Cells
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亚油酸 (9t, 12t-C18:2) 诱导人脐静脉内皮细胞凋亡的 Caspase 通路
DOI：
10.1007/s11745-012-3728-4
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发表时间：
2013-02-01
期刊：
影响因子：
1.9
通讯作者：
Du, Fang-Ling
中科院分区：
文献类型：
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作者：
Bin, Qiu;Rao, Huan;Du, Fang-Ling
据报道，反式脂肪酸（TFA）会导致炎症和冠心病。本研究旨在探讨两个双键TFA（TDTFA）对人脐静脉内皮细胞（HUVEC）的促凋亡作用。 HUVEC 在含有 50、100、200、400 μmol/l 的亚油酸 (9t,12t-C18:2) 的培养基中生长 24 或 48 小时，以检查 TDTFA 对这些细胞的活力和凋亡的影响。使用流式细胞术分析和共聚焦扫描来测量细胞凋亡、膜联蛋白V的细胞结合和碘化丙啶的摄取。采用比色法和RT-PCR分析HUVEC中caspase-3、-8和-9的酶活性和mRNA表达。结果表明，9t,12t-C18:2 以剂量依赖性和时间依赖性的方式抑制 HUVEC 的活力。与对照组相比，9t,12t-C18:2诱导的凋亡和坏死细胞的百分比显着增加。与对照组相比，9t、12t-C18:2 处理的细胞中 caspase-8、‐9 和 ‐3 的活性和 mRNA 表达显着增加。研究发现，在 HUVEC 中添加 caspase-8 (z-IETD-fmk) 和 caspase-9 (z-LEHD-fmk) 特异性抑制剂可完全抑制 9t,12t-C18:2 诱导的 caspase-3 激活，z-IETD-fmk 抑制 caspase-9 的激活。同时发现9t,12t-C18:2处理显着升高了Bid、Smac/DIABLO的mRNA表达和线粒体细胞色素c的释放。这些结果表明9t,12t-C18:2可能通过激活caspase-8、-9和-3诱导HUVEC凋亡。死亡受体途径和线粒体途径均可能参与9t,12t-C18:2诱导的细胞凋亡。
Trans fatty acids (TFA) are reported to contribute to inflammation and coronary heart disease. The study aim was to investigate the proapoptotic effects of two double bond TFA (TDTFA) on human umbilical vein endothelial cells (HUVEC). The HUVEC were grown in media supplied with linoelaidic acid (9t,12t‐C18:2) at 50, 100, 200, 400 μmol/l for 24 or 48 h to examine the effects of TDTFA on the viability and apoptosis of these cells. Flow cytometry analysis and confocal scanning were used to measure apoptosis, cell binding of Annexin V and propidium iodide uptake. Colorimetric assay and RT‐PCR were used to analyze enzyme activities and mRNA expression of caspase‐3, ‐8 and ‐9 in HUVEC. Results showed that 9t,12t‐C18:2 inhibited the viability of HUVEC in a dose‐dependent and time‐dependent manner. The percentages of 9t,12t‐C18:2 induced apoptotic and necrotic cells significantly increased compared with that of the control. The activities and mRNA expression of caspase‐8, ‐9 and ‐3 were significantly increased in 9t,12t‐C18:2 treated cells compared to that of the control. Addition of specific inhibitors of caspase‐8 (z‐IETD‐fmk) and caspase‐9 (z‐LEHD‐fmk) to HUVEC was found to completely inhibit 9t,12t‐C18:2‐induced activation of caspase‐3, and z‐IETD‐fmk inhibited the activation of caspase‐9. Meanwhile, it was found that mRNA expression of Bid, Smac/DIABLO and the release of mitochondrial cytochrome c were significantly elevated by 9t,12t‐C18:2 treatment. These results suggest that 9t,12t‐C18:2 may induce apoptosis of HUVEC through activating caspase‐8, ‐9 and ‐3. Both the death receptor pathway and the mitochondrial pathway may be involved in the apoptosis induced by 9t,12t‐C18:2.