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[正常小鼠血清通过抑制粘着斑信号通路减轻小鼠放射性肺炎]

[Normal mouse serum alleviates radiation pneumonitis in mice by inhibiting the focal adhesion signaling pathway].

作者信息

Yuan Tong, Guo Yuying, Zhang Junling, Fan Saijun

机构信息

Institute of Radiation Medicine, Chinese Academy of Medical Science//Peking Union Medical College; Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Tianjin 300192, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2024 May 20;44(5):801-809. doi: 10.12122/j.issn.1673-4254.2024.05.01.

Abstract

OBJECTIVE

To evaluate the therapeutic effect of normal mouse serum on radiation pneumonitis in mice and explore the possible mechanism.

METHODS

Mouse models of radiation pneumonitis induced by thoracic radiation exposure were given intravenous injections of 100 μL normal mouse serum or normal saline immediately after the exposure followed by injections once every other day for a total of 8 injections. On the 15th day after irradiation, histopathological changes of the lungs of the mice were examined using HE staining, the levels of TNF-α, TGF-β, IL-1α and IL-6 in the lung tissue and serum were detected using ELISA, and the percentages of lymphocytes in the lung tissue were analyzed with flow cytometry. High-throughput sequencing of exosome miRNA was carried out to explore the changes in the signaling pathways. The mRNA expression levels of the immune-related genes were detected by qRT-PCR, and the protein expressions of talin-1, tensin2, FAK, vinculin, α-actinin and paxillin in the focal adhesion signaling pathway were detected with Western blotting.

RESULTS

In the mouse models of radiation pneumonitis, injections of normal mouse serum significantly decreased the lung organ coefficient, lowered the levels of TNF-α, TGF-β, IL-1α and IL-6 in the serum and lung tissues, and ameliorated infiltration of CD45, CD4 and T lymphocytes in the lung tissue (all <0.05). The expression levels of and genes at both the mRNA and protein levels and the protein expressions of talin-1, tensin2, FAK, vinculin, α‑actinin and paxillin were all significantly down-regulated in the mouse models after normal mouse serum treatment.

CONCLUSION

Normal mouse serum ameliorates radiation pneumonitis in mice by inhibiting the expressions of key proteins in the Focal adhesion signaling pathway.

摘要

目的

评估正常小鼠血清对小鼠放射性肺炎的治疗效果并探讨其可能机制。

方法

对胸部接受辐射照射诱导放射性肺炎的小鼠模型,在照射后立即静脉注射100 μL正常小鼠血清或生理盐水,之后每隔一天注射一次,共注射8次。照射后第15天,采用苏木精-伊红(HE)染色检查小鼠肺部的组织病理学变化,用酶联免疫吸附测定(ELISA)法检测肺组织和血清中肿瘤坏死因子-α(TNF-α)、转化生长因子-β(TGF-β)、白细胞介素-1α(IL-1α)和白细胞介素-6(IL-6)的水平,用流式细胞术分析肺组织中淋巴细胞的百分比。进行外泌体微小核糖核酸(miRNA)的高通量测序以探索信号通路的变化。通过实时定量聚合酶链反应(qRT-PCR)检测免疫相关基因的信使核糖核酸(mRNA)表达水平,用蛋白质免疫印迹法检测粘着斑信号通路中踝蛋白-1(talin-1)、张力蛋白2(tensin2)、黏着斑激酶(FAK)、纽蛋白(vinculin)、α-辅肌动蛋白(α-actinin)和桩蛋白(paxillin)的蛋白表达。

结果

在放射性肺炎小鼠模型中,注射正常小鼠血清显著降低了肺器官系数,降低了血清和肺组织中TNF-α、TGF-β、IL-1α和IL-6的水平,并改善了肺组织中CD45、CD4和T淋巴细胞的浸润(均P<0.05)。正常小鼠血清处理后的小鼠模型中, 和 基因在mRNA和蛋白水平的表达以及talin-1、tensin2、FAK、vinculin、α-actinin和paxillin的蛋白表达均显著下调。

结论

正常小鼠血清通过抑制粘着斑信号通路中关键蛋白的表达改善小鼠放射性肺炎。

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