羊水间充质干细胞的条件培养基可以旁分泌方式调节人神经母细胞瘤细胞系 SY-SY5Y 的阿尔茨海默病样变化。

Conditioned medium from amniotic fluid mesenchymal stem cells could modulate Alzheimer's disease-like changes in human neuroblastoma cell line SY-SY5Y in a paracrine manner.

机构信息

Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Basic Medical Sciences, Khoy University of Medical Sciences, Khoy, Iran; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.

Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Tissue Cell. 2022 Jun;76:101808. doi: 10.1016/j.tice.2022.101808. Epub 2022 May 4.

DOI:10.1016/j.tice.2022.101808

PMID:35526311

Abstract

BACKGROUND

Alzheimer's disease is usually diagnosed by significant extracellular deposition of beta-amyloid and intracellular neurofibrillary tangle formation. Here, we investigated the paracrine effect of amniotic fluid-derived mesenchymal stem cells on AD changes in human SH-SY5Y cells.

METHODS

SH-SY5Y cells were divided into five groups: Control, 0.1 µg/ml LPS, 10 µg/ml LPS, 0.1 µg/ml LPS + conditioned medium, and 10 µg/ml LPS + conditioned medium. Cells were incubated with 0.1% and 10 µg/ml LPS for 48 h, followed by incubation with the conditioned medium of amniotic fluid-derived mesenchymal stem cells for the next 24 h. Beta-amyloid plaques were monitored by Congo-red staining. Survival and apoptosis were assessed by the MTT assay and flow cytometric analysis of Annexin-V. ELISA was used to measure the levels of neprilysin, angiotensin-converting enzyme, and Matrix Metalloproteinase-9. A PCR array was used to measure the expression of genes involved in neurogenesis.

RESULTS

Bright-field imaging showed beta-amyloid plaques in the group treated with 10 µg/ml LPS. We found minimal effects in groups receiving 0.1 µg/ml LPS. The data showed that the reduction in the levels of neprilysin, angiotensin-converting enzyme, and Matrix Metalloproteinase-9 in the LPS-treated cells was attenuated after incubation with the stem cell secretome (p < 0.05). Amniotic fluid stem cell secretome increased the viability of LPS-treated SH-SY5Y cells (p 0.05) and was associated with a decrease in apoptotic changes (p < 0.05). We found the modulation of several genes involved in neurogenesis in the 10 µg/ml LPS + conditioned medium group compared to cells treated with 10 µg/ml LPS alone.

CONCLUSION

Amniotic fluid stem cell secretion reduces AD-like pathologies in the human neuronal lineage.

摘要

背景

阿尔茨海默病通常通过β-淀粉样蛋白的显著细胞外沉积和细胞内神经纤维缠结的形成来诊断。在这里，我们研究了羊水衍生的间充质干细胞对人 SH-SY5Y 细胞 AD 变化的旁分泌作用。

方法

将 SH-SY5Y 细胞分为五组：对照组、0.1μg/ml LPS 组、10μg/ml LPS 组、0.1μg/ml LPS+条件培养基组和 10μg/ml LPS+条件培养基组。用 0.1%和 10μg/ml LPS 孵育细胞 48 小时，然后用羊水衍生的间充质干细胞的条件培养基孵育 24 小时。通过刚果红染色监测β-淀粉样斑块。通过 MTT 测定和 Annexin-V 流式细胞术分析评估细胞存活率和凋亡。ELISA 用于测量 Neprilysin、血管紧张素转换酶和基质金属蛋白酶-9 的水平。PCR 阵列用于测量参与神经发生的基因的表达。

结果

明场成像显示用 10μg/ml LPS 处理的组中存在β-淀粉样斑块。我们发现用 0.1μg/ml LPS 处理的组影响较小。数据显示，在用 LPS 处理的细胞中，Neprilysin、血管紧张素转换酶和基质金属蛋白酶-9 的水平降低，在用干细胞分泌液孵育后得到缓解（p<0.05）。羊水干细胞分泌液增加了 LPS 处理的 SH-SY5Y 细胞的活力（p<0.05），并与凋亡变化减少有关（p<0.05）。与单独用 10μg/ml LPS 处理的细胞相比，我们发现 10μg/ml LPS+条件培养基组中几个参与神经发生的基因发生了调节。