整合素 α5β1 抑制 rBMSCs 失巢凋亡并促进一氧化氮的产生。
Integrin alpha5beta1 suppresses rBMSCs anoikis and promotes nitric oxide production.
机构信息
Central laboratory, and key laboratory of Oral and Maxillofacial-Head and Neck Medical Biology, Liaocheng People's Hospital, Liaocheng, Shandong, 252000, China.
Department of Cardiology, Liaocheng People's Hospital and Clinical School of Taishan Medical University, Liaocheng, Shandong, 252000, China.
出版信息
Biomed Pharmacother. 2018 Mar;99:1-8. doi: 10.1016/j.biopha.2018.01.038. Epub 2018 Jan 8.
BACKGROUND
Cell based therapy has been heralded as a novel, promising therapeutic strategy for cardiovascular diseases including pulmonary arterial hypertension (PAH). However, the low survival rate after transplantation due to cell death via anoikis is a major obstacle in stem cell therapy. Cells adhesion via Integrin alpha5beta1 (ITGA5B1) has a tendency to exert higher maximum forces. The present study aimed to evaluate the potential protective effect of ITGA5B1 on rat bone marrow mesenchymal stem cells (rBMSCs) from anoikis.
METHODS
Mononuclear cells were isolated by density gradient centrifugation with Ficoll, and rBMSCs cell surface markers were evaluated by flow cytometry. Osteogenic and adipocyte differentiation was determined by Alizarin Red S and Oil Red O staining respectively. The expression of Integrin A5 (ITGA5), Integrin B1 (ITGB1), eNOS and actived-caspase-3 mRNA or protein was confirmed by qPCR and western-blot. Cell adhesion, cell viability, anoikis and the migration of rBMSCs were also evaluated. Nitric oxide (NO) production was detected by the greiss assay.
RESULTS
Co-infected with Integrin A5 and B1 lentivirus to rBMSCs increased ITGA5 and ITGB1 mRNA and protein expression. ITGA5B1 enhanced the cell adhesion, cell viability, cell migration and NO production but reduced the cell anoikis in rBMSCs/ITGA5B1 groups.
CONCLUSION
Transduction of rat rBMSCs with ITGA5B1 lentivirus could prevent cell anoikis and increase NO production.
背景
细胞治疗被认为是一种新颖的、有前途的治疗策略，可用于治疗心血管疾病，包括肺动脉高压（PAH）。然而，由于细胞凋亡导致移植后存活率低，这是干细胞治疗的一个主要障碍。细胞通过整合素 alpha5beta1（ITGA5B1）黏附往往会产生更高的最大力。本研究旨在评估 ITGA5B1 对大鼠骨髓间充质干细胞（rBMSCs）抗细胞凋亡的潜在保护作用。
方法
通过密度梯度离心用 Ficoll 分离单核细胞，并通过流式细胞术评估 rBMSCs 细胞表面标志物。通过茜素红 S 和油红 O 染色分别确定成骨细胞和脂肪细胞分化。通过 qPCR 和 Western blot 验证整合素 A5（ITGA5）、整合素 B1（ITGB1）、eNOS 和活化 caspase-3 mRNA 或蛋白的表达。还评估了 rBMSCs 的细胞黏附、细胞活力、细胞凋亡和迁移。通过格里塞斯测定法检测一氧化氮（NO）的产生。
结果
共感染整合素 A5 和 B1 慢病毒到 rBMSCs 增加了 ITGA5 和 ITGB1 的 mRNA 和蛋白表达。ITGA5B1 增强了 rBMSCs/ITGA5B1 组的细胞黏附、细胞活力、细胞迁移和 NO 产生，但减少了细胞凋亡。
结论
用 ITGA5B1 慢病毒转导大鼠 rBMSCs 可防止细胞凋亡并增加 NO 产生。
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