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首页> 外文期刊>Life sciences >27-Hydroxycholesterol inhibits neutral sphingomyelinase in cultured human endothelial cells.
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27-Hydroxycholesterol inhibits neutral sphingomyelinase in cultured human endothelial cells.

机译：27-羟基胆固醇抑制培养的人内皮细胞中的中性鞘磷脂酶。

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To study the effect of 27-hydroxycholesterol (27OHC) on the catabolism of sphingomyelin, we cultured endothelial cells (ECs) from human umbilical veins with 27OHC, then measured activities of acid sphingomyelinase (ASMase) and neutral sphingomyelinase (NSMase) and sphingomyelin consumption by using [14C]sphingomyelin, and determined NSMase mRNA expressions by RT-PCR method. The results indicated that [14C]sphingomyelin accumulated in cells treated with 27OHC, and that the activities of both NSMase and ASMase were inhibited in ECs cultured with 27OHC. To further study the effect of 27OHC on NSMase, we used desipramine, an inhibitor of ASMase, to exclude the possible interference of ASMase's residual activity at neutral condition. Also, we observed the significant inhibition of NSMase activity by using glutathione, an inhibitor of NSMase, but found no further impact when 27OHC was added later. To determine whether the inhibition of NSMase activity was directly due to the effect of 27OHC, we exposed cell homogenate to 27OHC, and found no inhibitive effect of 27OHC on the activity of NSMase. All of our data confirmed that 27OHC had only an indirect inhibitive effect on NSMase. Our finding that no change of the NSMase mRNA expression by 27OHC indicated that the inhibitive effect of 27OHC on NSMase activity occurred at a post-transcriptional level. We suggest that an altered membrane fluidity caused by 27OHC could be involved in the inhibited activity of NSMase.
机译：为了研究27-羟基胆固醇（27OHC）对鞘磷脂分解代谢的影响，我们用27OHC培养了人脐静脉内皮细胞（EC），然后测量了酸性鞘磷脂酶（ASMase）和中性鞘磷脂酶（NSMase）的活性并通过鞘磷脂的消耗使用[14C]鞘磷脂，并通过RT-PCR方法测定NSMase mRNA表达。结果表明，[14C]鞘磷脂在27OHC处理的细胞中积累，并且NSMase和ASMase的活性在27OHC培养的EC中均受到抑制。为了进一步研究27OHC对NSMase的影响，我们使用了desipramine（ASMase的抑制剂）排除了中性条件下ASMase残留活性的可能干扰。此外，我们观察到通过使用NSMase抑制剂谷胱甘肽对NSMase活性的显着抑制，但后来添加27OHC时没有发现进一步的影响。为了确定NSMase活性的抑制作用是否直接归因于27OHC的作用，我们将细胞匀浆暴露于27OHC，未发现27OHC对NSMase活性的抑制作用。我们所有的数据都证实27OHC对NSMase仅具有间接抑制作用。我们的发现：27OHC对NSMase mRNA表达没有变化，表明27OHC对NSMase活性的抑制作用发生在转录后水平。我们建议由27OHC引起的膜流动性改变可能与NSMase的抑制活性有关。

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