Ding Hongfang, Zhang Hui, Ding Huifang, Li Dong, Yi Xinhao, Ma Xiaoxu, Li Ruijuan, Huang Mei, Ju Xiuli
Department of Pediatrics, Shengli Oil field Central Hospital, 31 Jinan Road, Dongying, Shandong, China.
Department of Neurology, Qingdao Municipal Hospital, the Affiliated Hospital of Medical College of Qingdao University, 1 Jiaozhou Road, Qingdao, Shandong, China.
Cell Mol Immunol. 2017 Aug;14(8):693-701. doi: 10.1038/cmi.2015.99. Epub 2015 Dec 28.
Hypoxic-ischemic brain damage (HIBD) is a common cause of infant death. The purpose of our research was to explore the immunoregulatory mechanism of placenta-derived mesenchymal stem cells (PD-MSCs) in HIBD treatment. Seven-day-old rat pups were randomly divided into HIBD, PD-MSC, fibroblast, and control groups. Forty-eight hours after HIBD induction, cells at a density of 5 × 104 cells/10 µl were injected into the cerebral tissue in the PD-MSC and fibroblast groups. The TNF-α, interleukin- 17 (IL-17), interferon-γ (IFN-γ), and IL-10 levels were detected through quantitative real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Regulatory T cell (Tregs) populations were detected through flow cytometry, and forkhead box P3 (Foxp3) was measured through western blot analysis. Behavioral tests and gross and pathological examinations showed that PD-MSC treatment exerted significantly stronger neuroprotective effects than the other treatments. The expression levels of pro-inflammatory cytokines were substantially upregulated after HI injury. Compared with fibroblast treatment, PD-MSC treatment inhibited the production of pro-inflammatory cytokines and increased the production of IL-10 in the ischemic hemispheres and peripheral blood serum (all P < 0.01). Flow cytometry results showed a notable increase in the number of Tregs within the spleen of the HIBD group. Moreover, the number of Tregs and the Foxp3 expression levels were higher in the PD-MSC treatment group than in the HIBD and fibroblast groups (all P < 0.01). Our research suggests that the mechanism of PD-MSC treatment for HIBD partially involves inflammatory response suppression.
缺氧缺血性脑损伤(HIBD)是婴儿死亡的常见原因。我们研究的目的是探讨胎盘来源的间充质干细胞(PD-MSCs)在HIBD治疗中的免疫调节机制。将7日龄大鼠幼崽随机分为HIBD组、PD-MSC组、成纤维细胞组和对照组。诱导HIBD后48小时,将密度为5×104个细胞/10微升的细胞注射到PD-MSC组和成纤维细胞组的脑组织中。通过定量实时聚合酶链反应(RT-PCR)和酶联免疫吸附测定(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-17(IL-17)、干扰素-γ(IFN-γ)和IL-10水平。通过流式细胞术检测调节性T细胞(Tregs)群体,并通过蛋白质印迹分析测定叉头框P3(Foxp3)。行为测试以及大体和病理检查表明,PD-MSC治疗比其他治疗具有更强的神经保护作用。HI损伤后促炎细胞因子的表达水平显著上调。与成纤维细胞治疗相比,PD-MSC治疗抑制了缺血半球和外周血血清中促炎细胞因子的产生,并增加了IL-10的产生(所有P<0.01)。流式细胞术结果显示,HIBD组脾脏内Tregs数量显著增加。此外,PD-MSC治疗组的Tregs数量和Foxp3表达水平高于HIBD组和成纤维细胞组(所有P<0.01)。我们的研究表明,PD-MSC治疗HIBD的机制部分涉及炎症反应抑制。