PHISIOLOGICAL ACTIVITY OF CERAMIDE AND RELATED SUBSTANCE

神经酰胺及相关物质的生理活性

• 批准号: 07672357
• 负责人: MASUZAWA Yasuo
• 金额: $ 1.41万
• 依托单位: HYOGO UNIVERSITY OF TEACHER EDUCATION
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07672357/
• 关键词: ceramide; cis-trans isomer; acetylene type; molecular species; quantification; apoptosis; docosahexaenoic acid; sphingosine; 新規定量法

1. A new method for quantitative analysis of ceramide molecular species was developed using fluorescence HPLC.This method was applied to quantification of free ceramide in HL60 and U937 cells. Total amount of free ceramide was 3.9 <plus-minus> 0.42 mug/108 cells and 2.6 <plus-minus> 0.43 mug/108 cells, respectively. This is the first quantitative analysis of molecular species of free ceramide in intact ce2. The requirement of a trans double bond for the biological action of ceramide was assessed by comparing the apoptosis-inducing activity of various ceramide analogs. An acetylene type derivative (TRP-Cer) yielded the highest percentage of apoptotic cells corresponding to three times that induced by C6-Cer. C6-cis-Cer also showed stronger activity than C6-Cer. These observations suggest that the trans configuration of ceramide is not necessarily essential for the activity to induce apoptosis. In addition, distinctive activity of C6-TRP-Cer suggests that this ceramide analog might be useful for developing a new type of antitumor drug.3. Effect of exogenously-added polyunsaturated fatty acids on the apoptosis of HL60 induced by cell-permeable ceramide or sphingosine. Ceramide-induced apoptosis was not affected by the preincubation with polyunsaturated fatty acids. However sphingosine-induced apoptosis was much attenuated by 24h incubation with DHA.These results indicated that the mechanism for the induction of apoptosis was different between ceramide and sphingosine, and that DHA may specifically reduce PKC inhibition-dependent apoptosis.

1.建立了一种新的神经酰胺分子种类定量分析方法，并将该方法应用于HL60和U937细胞中游离神经酰胺的定量。游离神经酰胺总量分别为3.9正负0.42µg/108细胞和2.6正负0.43µg/108细胞。这是首次对完整CE2中游离神经酰胺的分子种类进行定量分析。通过比较不同神经酰胺类似物的诱导凋亡活性，评估了神经酰胺生物学作用所需的反式双键。乙炔类衍生物(Trp-Cer)诱导的细胞凋亡率最高，相当于C6-Cer的3倍。C6-cis-Cer的活性也高于C6-Cer。这些观察表明神经酰胺的反式构型不一定是诱导细胞凋亡的活性所必需的。此外，C6-Trp-Cer的独特活性表明该神经酰胺类似物可能有助于开发新型的抗肿瘤药物。外源性多不饱和脂肪酸对神经酰胺或鞘氨醇诱导HL60细胞凋亡的影响。神经酰胺诱导的细胞凋亡不受多不饱和脂肪酸预孵育的影响。神经酰胺和鞘氨醇诱导细胞凋亡的机制不同，DHA可能特异性地抑制PKC抑制的细胞凋亡。

项目成果

Etsu Kishida et al.: "Evaluation of a trans contignlation for the apoptosis-inducing activity of cerawide" J. Lipid Mediators Cell Signaling. (印刷中). (1997)

Etsu Kishida 等人：“cerawide 细胞凋亡诱导活性的反式连接的评估”J.Lipid Mediators Cell Signaling（正在出版）。