Philosophy Doctor in Medicine Program, Faculty of Medicine, Universitas Sumatera Utara, Medan, Indonesia.
Department of Internal Medicine, Faculty of Medicine, Universitas Sumatera Utara, Medan, Indonesia.
Med Arch. 2023 Apr;77(2):90-96. doi: 10.5455/medarh.2023.77.90-96.
Lower limb peripheral artery disease (PAD) is the main risk of diabetes mellitus which result to high mortality rate. Approximately, 50% of patients who receive several treatments have passed away or lost limbs at a year's follow-up. Secretome of hypoxia mesenchymal stem cells (S-MSCs) contains several active soluble molecules from hypoxia MSCs (H-MSCs) that capable inducing anti-inflammatory and vascular regeneration in PAD.
In this study, we investigated the therapeutic potential of S-MSCs in improving dynamic function and angiogenesis of PAD diabetic rats.
The PAD was established by the incision from the groin to the inner thigh and distal ligation of femoral arteries in rats with diabetes. Rats were administered with 200 µL and 400 µL S-MSCs that successfully filtrated using tangential flow filtration (TFF) system based on various molecular weight cut-off categories intravenously. ELISA assay was used to analyze the cytokines and growth factors contained in S-MSCs. Tarlov score were examined at day 1, 3, 5, 7, 10 and 14. The rats were sacrificed at day 14 and muscle tissues were collected for immunohistochemistry (IHC) and gene expression analysis.
ELISA assay showed that S-MSCs provides abundant level of VEGF, PDGF, bFGF, IL-10 and TGFβ. In vivo administration of S-MSCs remarkably enhanced the Tarlov score. S-MSCs improved angiogenesis through enhancing VEGF gene expression and significantly increasing CD31 positive area in muscle tissue of PAD diabetic rats.
Our findings suggest that S-MSCs could improves dynamic function and angiogenesis in PAD diabetic rats.
下肢外周动脉疾病（PAD）是糖尿病的主要风险因素，导致死亡率较高。大约 50%的接受多种治疗的患者在一年的随访中已经死亡或失去肢体。低氧间充质干细胞的分泌组（S-MSCs）包含来自低氧 MSCs（H-MSCs）的几种活性可溶性分子，能够在 PAD 中诱导抗炎和血管再生。
本研究旨在探讨 S-MSCs 改善 PAD 糖尿病大鼠运动功能和血管生成的治疗潜力。
通过在糖尿病大鼠的腹股沟到大腿内侧和股动脉远端结扎切口建立 PAD。大鼠通过静脉内给予成功过滤的 S-MSCs（使用切向流过滤（TFF）系统基于各种分子量截止类别），剂量为 200µL 和 400µL。ELISA 测定法用于分析 S-MSCs 中包含的细胞因子和生长因子。在第 1、3、5、7、10 和 14 天检查 Tarlov 评分。在第 14 天处死大鼠并收集肌肉组织进行免疫组织化学（IHC）和基因表达分析。
ELISA 测定法显示 S-MSCs 提供了丰富的 VEGF、PDGF、bFGF、IL-10 和 TGFβ。体内给予 S-MSCs 可显著提高 Tarlov 评分。S-MSCs 通过增强 VEGF 基因表达和显著增加 PAD 糖尿病大鼠肌肉组织中 CD31 阳性面积来改善血管生成。
我们的研究结果表明，S-MSCs 可以改善 PAD 糖尿病大鼠的运动功能和血管生成。
