Larasati Vinny A, Lembang Gregorius V, Tjahjono Yudy, Winarsih Sugi, Ana Ika Dewi, Wihadmadyatami Hevi, Kusindarta Dwi L
Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia.
Biomedical Laboratory, Faculty of Pharmacy, Widya Mandala Catholic University, Surabaya 60112, Indonesia.
Vet Sci. 2022 Jan 27;9(2):48. doi: 10.3390/vetsci9020048.
Mesenchymal stem cells (MSCs) and conditioned medium (CM) derived from human umbilical blood cord stem cells (HUBSC) are now being extensively utilized. Human umbilical vein endothelial cells (HUVECs) have the same ability as HUBSC as an option for autologous therapy. In addition, cell therapy using HUVECs may produce protective signals for cerebral vessels and promote neuronal survival after hypoxic-ischemic damage. HUVECs have the same anatomical and physiological structure as bovine umbilical vein endothelial cells (BUVECs). In this study, we aim to determine the ability of BUVEC-CM to reduce inflammation and apoptosis on in vitro neurodegeneration models (PC12 and SH-SY5Y cell lines). BUVEC-CM obtained from the third and fourth passages were analyzed using liquid chromatography-mass spectrometry (LC-MS) and high-resolution mass spectrometry (HR-MS), while the other part was used as a treatment for in vitro model neurodegeneration. The PC12 and SH-SY5Y cell lines were cultured and grouped into seven different treatments, including untreated cells. As the treatment group, cells were given TMT 10 µM in the presence of different doses of CM (25%, 50%, 75%, and 100%); as a control comparison of recent therapy, donepezil was used. In addition, cells with the administration of TMT 10 µM were run as a positive control. Cell viability assay (CCK-8) and enzyme-linked immunosorbent assay (ELISA) were performed to identify the viability and expression of interleukin-1β (IL-1β), caspase-3, and caspase-9 for both PC12 and SH-SY5Y cells. The results showed that BUVEC-CM could significantly reduce IL-1β expression and downregulate caspase-3 and caspase-9, as well as when compared to the donepezil group. Taken together, these results indicate that BUVEC-CM can be used as a potential candidate for neuroprotective agents by reducing the activity of IL-1β and the expression of caspase-9 and caspase-3 in PC12 and SH-SY5Y cells induced by TMT. However, further research still needs to be conducted.
间充质干细胞（MSCs）和源自人脐带血干细胞（HUBSC）的条件培养基（CM）目前正在被广泛应用。人脐静脉内皮细胞（HUVECs）作为自体治疗的一种选择，具有与HUBSC相同的能力。此外，使用HUVECs进行细胞治疗可能会产生脑血管保护信号，并促进缺氧缺血性损伤后神经元的存活。HUVECs与牛脐静脉内皮细胞（BUVECs）具有相同的解剖和生理结构。在本研究中，我们旨在确定BUVEC-CM在体外神经退行性模型（PC12和SH-SY5Y细胞系）上减轻炎症和凋亡的能力。对从第3代和第4代获得的BUVEC-CM进行液相色谱-质谱联用（LC-MS）和高分辨率质谱（HR-MS）分析，另一部分用作体外神经退行性模型的治疗。将PC12和SH-SY5Y细胞系进行培养，并分为七种不同的处理组，包括未处理的细胞。作为处理组，在不同剂量的CM（25%、50%、75%和100%）存在的情况下，给予细胞10 µM的四甲基秋水仙碱（TMT）；作为近期治疗的对照比较，使用多奈哌齐。此外，给予10 µM TMT的细胞作为阳性对照。进行细胞活力测定（CCK-8）和酶联免疫吸附测定（ELISA），以鉴定PC12和SH-SY5Y细胞中白细胞介素-1β（IL-1β）、半胱天冬酶-3和半胱天冬酶-9的活力和表达。结果表明，与多奈哌齐组相比，BUVEC-CM能显著降低IL-1β的表达，并下调半胱天冬酶-3和半胱天冬酶-9的表达。综上所述，这些结果表明，BUVEC-CM通过降低TMT诱导的PC12和SH-SY5Y细胞中IL-1β的活性以及半胱天冬酶-9和半胱天冬酶-3的表达，可作为神经保护剂的潜在候选物。然而，仍需要进行进一步的研究。
