Department of Histology and Cell Biology, Faculty of Medicine, Zagazig University, Asharquia, Zagazig, 44519, Egypt.
Department of Medical Biochemistry, Faculty of Medicine, Zagazig University, Zagazig, Egypt.
Cell Tissue Res. 2018 Oct;374(1):83-97. doi: 10.1007/s00441-018-2838-6. Epub 2018 Apr 23.
Being one of the most debilitating complications among diabetic patients, diabetic polyneuropathy (DPN) is a paramount point of continuous research. Stem cell therapies have shown promising results. However, limited cell survival and paracrine activities hinder its transfer from bench to bedside. We designed this study to evaluate fluoxetine-pretreatment technique of mesenchymal stem cells (MSCs) as an approach to enhance their paracrine and immunomodulatory properties in DPN. Effects of fluoxetine treatment of MSCs were tested in vitro. Forty-two adult Wistar male albino rats were utilized, further subdivided into control, diabetic, MSC-treated and fluoxetine-pretreated MSC groups. Sciatic nerve sections were prepared for light and electron microscope examination and immunohistochemical detection of neurofilament (NF) protein. Also, we assessed in vitro survival and paracrine properties of fluoxetine-pretreated MSCs. Real time PCR of BDNF, VEGF, IL-1β, and IL-10 expression in tissue homogenate was performed. Our results showed restoration of normal neuronal histomorphology and ultrastructure, moreover, immunohistochemical expression of anti-neurofilament protein was significantly elevated in MSC-treated groups compared to the diabetic one. Fluoxetine enhanced the MSC survival and their paracrine properties of MSCs in vitro. Furthermore, the fluoxetine-pretreated MSC group revealed a significant elevation of mRNA expression of BDNF (neurotrophic factor) and VEGF (angiogenic factor), denoting ameliorated MSC paracrine properties. Similarly, improved immunomodulatory functions were evident by a significant reduction of interleukin-1β mRNA expression (pro-inflammatory) and a reciprocal significant increase of interleukin-10 (anti-inflammatory). We concluded that fluoxetine-pretreatment of MSCs boosts their survival, paracrine, and immunomodulatory traits and directly influenced neuronal histomorphology. Hence, it presents a promising intervention of diabetic polyneuropathy. Graphical Abstract.
作为糖尿病患者中最具致残性的并发症之一，糖尿病多发性神经病（DPN）是一个持续研究的重点。干细胞疗法已显示出良好的效果。然而，有限的细胞存活和旁分泌活性阻碍了其从实验室到临床的转化。我们设计了这项研究，以评估氟西汀预处理间充质干细胞（MSCs）的技术，作为增强其在 DPN 中旁分泌和免疫调节特性的方法。在体外测试了氟西汀处理 MSCs 的效果。使用 42 只成年雄性 Wistar 白化大鼠，进一步分为对照组、糖尿病组、MSC 治疗组和氟西汀预处理 MSC 组。制备坐骨神经切片，进行光镜和电镜检查以及神经丝（NF）蛋白的免疫组织化学检测。此外，我们评估了氟西汀预处理 MSC 的体外存活和旁分泌特性。对组织匀浆中 BDNF、VEGF、IL-1β 和 IL-10 表达进行实时 PCR。我们的结果表明，氟西汀预处理恢复了正常的神经元形态和超微结构，此外，与糖尿病组相比，MSC 治疗组的抗神经丝蛋白免疫组织化学表达显著升高。氟西汀增强了 MSC 的存活及其在体外的旁分泌特性。此外，氟西汀预处理 MSC 组 BDNF（神经营养因子）和 VEGF（血管生成因子）的 mRNA 表达显著升高，表明 MSC 旁分泌特性得到改善。同样，白细胞介素-1β mRNA 表达（促炎）的显著降低和白细胞介素-10（抗炎）的显著增加表明免疫调节功能得到改善。我们得出结论，氟西汀预处理 MSC 可增强其存活、旁分泌和免疫调节特性，并直接影响神经元形态。因此，它为糖尿病多发性神经病提供了一种有前途的干预方法。
