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人骨髓间充质干细胞对过敏性哮喘小鼠气道炎症的影响及其机制

[Effects of human mesenchymal stem cells on airway inflammation in allergic asthma mice and the underlying mechanism].

作者信息

Ren N N, Chen H J, Chen Q L, Eileen J, An Y G, Lin Yiguang

机构信息

Guangdong Online Hospital, Guangdong 2nd Provincial People's Hospital, Guangzhou 510317, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2017 Sep 12;97(34):2697-2702. doi: 10.3760/cma.j.issn.0376-2491.2017.34.012.

Abstract

To investigate the effects of human umbilical cord mesenchymal stem cells (hUC-MSCs) on airway inflammation in an ovalbumin (OVA) induced asthma mouse model and the underlying mechanism. Twenty-four BALB/c mice were randomly divided into four equal groups: normal control group, OVA-induced asthmatic model group, hUC-MSCs treated group (50 μl of hUC-MSCs was transplanted into the trachea of asthmatic mice ) and hUC-MSCs control group (50 μl of hUC-MSCs was transplanted into the trachea of control mice). Human umbilical cord mesenchymal stem cells from umbilical cord of healthy new born babies were used as the source of hUC-MSCs for this study. The asthmatic conditions of the airways and the lungs were assessed by examining: (1) histopathological changes of the airways and the lungs; (2) expression of cytokines IL-6 and TGF-β mRNA by real-time PCR; (3) total leukocytes and mast cell count in bronchoalveolar lavage fluid (BALF) and number of IL-17-expressing CD4(+) cells (Th17 cells) in the lung tissue using flow cytometry. Typical histopathological changes of asthma were confirmed in the asthmatic model group. These changes included intensive inflammatory cell infiltration around the airways and patchy airway occlusion by hyperviscous mucus. The number of total leukocytes and mast cells in BALF were significantly increased in the asthmatic mice when compared with the control group (<0.05). Mice in the asthmatic model group had significantly higher percentage of Th17 cells in lung tissues when compared with the control group (2.90% vs 0.76%, <0.05). In contrast, in the asthmatic mice treated with hUC-MSCs, the inflammatory cell infiltration was significantly reduced compared with asthmatic mice, as observed by significantly lower leukocytes and mast cells in BALF (<0.05) and significant reduction in the percentage of Th17 cells in the lung of OVA-challenged mice following hUC-MSCs treatment (percentage of Th17 cells: 0.24% vs 2.90%, <0.05). The expression of mRNA for IL-6 and TGF-β was significantly suppressed in the hUC-MSCs treatment group (0.23 vs 2.30 and 0.56 vs 6.60, both <0.01). No asthmatic pathological changes in both normal and hUC-MSCs control groups were observed. hUC-MSCs significantly inhibit the airway inflammation in OVA-induced asthmatic mice. This inhibition is associated with the suppression of Th17 cells and the down-regulation of inflammatory factors such as IL-6 and TGF-β in the lungs.

摘要

探讨人脐带间充质干细胞(hUC-MSCs)对卵清蛋白(OVA)诱导的哮喘小鼠模型气道炎症的影响及其潜在机制。将24只BALB/c小鼠随机分为四组,每组6只:正常对照组、OVA诱导的哮喘模型组、hUC-MSCs治疗组(将50 μl hUC-MSCs移植到哮喘小鼠气管内)和hUC-MSCs对照组(将50 μl hUC-MSCs移植到对照小鼠气管内)。本研究采用健康新生儿脐带中的人脐带间充质干细胞作为hUC-MSCs的来源。通过以下方法评估气道和肺部的哮喘情况:(1)气道和肺部的组织病理学变化;(2)通过实时PCR检测细胞因子IL-6和TGF-β mRNA的表达;(3)使用流式细胞术检测支气管肺泡灌洗液(BALF)中的总白细胞和肥大细胞计数以及肺组织中表达IL-17的CD4(+)细胞(Th17细胞)的数量。哮喘模型组证实了典型的哮喘组织病理学变化。这些变化包括气道周围密集的炎性细胞浸润和高粘性黏液导致的斑片状气道阻塞。与对照组相比,哮喘小鼠BALF中的总白细胞和肥大细胞数量显著增加(<0.05)。与对照组相比,哮喘模型组小鼠肺组织中Th17细胞的百分比显著更高(2.90%对0.76%,<0.05)。相比之下,在接受hUC-MSCs治疗的哮喘小鼠中,与哮喘小鼠相比,炎性细胞浸润显著减少,BALF中的白细胞和肥大细胞显著降低(<0.05),并且hUC-MSCs治疗后OVA激发的小鼠肺中Th17细胞百分比显著降低(Th17细胞百分比:0.24%对2.90%,<0.05)。hUC-MSCs治疗组中IL-6和TGF-β的mRNA表达显著受到抑制(0.23对2.30以及0.56对6.60,均<0.01)。正常组和hUC-MSCs对照组均未观察到哮喘病理变化。hUC-MSCs显著抑制OVA诱导的哮喘小鼠的气道炎症。这种抑制与Th17细胞的抑制以及肺中炎性因子如IL-6和TGF-β的下调有关。

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