Kusindarta Dwi Liliek, Wihadmadyatami Hevi, Haryanto Aris
Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Department of Biochemistry, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Vet World. 2016 Dec;9(12):1348-1354. doi: 10.14202/vetworld.2016.1348-1354. Epub 2016 Dec 3.
This research was conducted to identify the expression of choline acetyltransferase (ChAT) in human cerebral microvascular endothelial cells (HCMECs) and to clarify the capability of Linn. ethanolic extract to stimulate the presence of ChAT in the aging HCMECs.
In this study, we perform an analysis some in the presence of an ethanolic extract of Linn. as a stimulator for the ChAT expression. HCMECs are divided become two groups, the first is in low passage cells as a model of young aged and the second is in a high passage as a model of aging. Furthermore to analysis the expression of ChAT without and with extract treatments, immunocytochemistry and flow cytometry analysis were performed. In addition, ChAT sandwich enzyme-linked immunosorbent assay is developed to detect the increasing activity of the ChAT under normal, and aging HCMECs on the condition treated and untreated cells.
In our models using HCMECs, we found that ChAT is expressed throughout intracytoplasmic areas. On the status of aging, the ethanolic extract from Linn. is capable to stimulate and restore the expression of ChAT. The increasing of ChAT expression is in line with the increasing activity of this enzyme on the aging treated HCMECs.
Our observation indicates that HCMECs is one of the noncholinergic cells which is produced ChAT. The administrated of Linn. ethanolic extract may stimulate and restore the expression of ChAT on the deteriorating cells of HCMECs, thus its may give nerve protection and help the production of acetylcholine.
本研究旨在确定胆碱乙酰转移酶(ChAT)在人脑海微血管内皮细胞(HCMECs)中的表达,并阐明[植物名称]乙醇提取物刺激衰老HCMECs中ChAT表达的能力。
在本研究中,我们在[植物名称]乙醇提取物作为ChAT表达刺激剂存在的情况下进行了一些分析。HCMECs被分为两组,第一组是低代细胞作为年轻细胞模型,第二组是高代细胞作为衰老模型。此外,为了分析有无提取物处理时ChAT的表达情况,进行了免疫细胞化学和流式细胞术分析。另外,开发了ChAT夹心酶联免疫吸附测定法来检测正常和衰老HCMECs在处理和未处理细胞条件下ChAT活性的增加。
在我们使用HCMECs的模型中,我们发现ChAT在整个细胞质区域表达。在衰老状态下,[植物名称]的乙醇提取物能够刺激并恢复ChAT的表达。ChAT表达的增加与该酶在衰老处理的HCMECs上活性的增加一致。
我们的观察表明HCMECs是产生ChAT的非胆碱能细胞之一。给予[植物名称]乙醇提取物可能会刺激并恢复HCMECs退化细胞中ChAT的表达,因此它可能提供神经保护并有助于乙酰胆碱的产生。