| 杭青青,应航洁,金佳男,方敏,包勇,陈明,彭芳.克唑替尼上调TGF-β1通路对小鼠急性放射性肺损伤的影响[J].中华放射医学与防护杂志,2022,42(6):408-415 |
| 克唑替尼上调TGF-β1通路对小鼠急性放射性肺损伤的影响 |
| Effect of crizotinib on TGF-β1 signaling pathway in acute radiation-induced lung injury in mice |
| 投稿时间：2021-10-08 |
| DOI：10.3760/cma.j.cn112271-20211008-00403 |
| 中文关键词: 放射性肺损伤 克唑替尼 小鼠 靶向药物 放疗 |
| 英文关键词:Radiation-induced lung injury Crizotinib Mice Targeted drugs Radiotherapy |
| 基金项目:国家自然科学基金(81672972, 81703018) |
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| 中文摘要: |
| 目的 研究克唑替尼对小鼠急性放射性肺损伤的影响及其可能机制。方法 应用随机数表法将72只小鼠分成4组：健康对照组(NC组)12只、单纯给药组(CRZ组)12只、单纯照射组(RT组)24只、克唑替尼+照射组(RT+CRZ组)24只。采用全肺单次12 Gy X射线照射。分别于照射后1、2、4、8周处死获取肺组织标本及肺泡灌洗液，光学显微镜下计数有核细胞的总数，聚氰基丙烯酸正丁酯(BCA)蛋白检测法检测蛋白质浓度，HE染色和MASSON染色观察肺组织病理变化，实时荧光定量(qPCR)检测转化生长因子β1(TGF-β1)、细胞间黏附分子1(ICAM-1)mRNA水平表达，免疫组织化学法观察髓过氧化物酶(MPO)和ICAM-1的定位和表达，Western blot法检测TGF-β1、Smad3、p-Smad3及ICAM-1蛋白表达情况。结果 在各个时间点上，RT+CRZ组肺组织炎症、渗出表现显著加重，肺泡灌洗液中的细胞总数及蛋白浓度均较其他3组增多，与RT组相比，第4周差异具有统计学意义(t=-5.031、-2.814，P < 0.05)。照射后第1、4、8周，与RT组相比，RT+CRZ组肺组织中的TGF-β1表达增高明显(t=-2.687、-7.032、-5.221，P < 0.05)，照射后第2、4周，RT+CRZ组ICAM-1表达增高明显(t=-4.819、-6.057，P < 0.05)，至第4周达峰，第8周时表达下降，同时，MPO的表达变化与ICAM-1、TGF-β1变化趋势一致。第4周时，4组小鼠Smad3蛋白表达差异无统计学意义(P>0.05)；RT+CRZ组小鼠肺组织内的TGF-β1、p-Smad3、ICAM-1蛋白含量及p-Smad3/Smad3均高于其他3组，差异具有统计学意义(F=14.74、10.03、35.29、22.94，P < 0.05)。结论 克唑替尼联合照射会加重急性放射性肺损伤，其机制可能与通过上调TGF-β1信号通路增加ICAM-1表达有关。 |
| 英文摘要: |
| Objective To study the effect of crizotinib on acute radiation-induced lung injury in mice and its possible mechanism.Methods A total of 72 mice were randomly divided into 4 groups by the random number table method: healthy control group (NC group, n=12), crizotinib-only group (CRZ group, n=12), radiotherapy-only group (RT group, n=24), and radiotherapy pluscrizotinib group (RT+CRZ group, n=24). The whole lungs were exposed to a single dose of 12 Gy X-rays. Lung tissue and bronchoalveolar lavage fluid (BALF) were obtained at 1, 2, 4, and 8 weeks after radiotherapy. The total number of nucleated cells was counted under a light microscope, and the total protein content of BALF was detected by bicinchoninic acid (BCA) protein assay. The pathological changes of lung tissue were observed by HE staining and MASSON staining. The expressions of TGF-β1 and ICAM-1 mRNA in lung tissue were detected by real-time quantitative polymerase chain reaction (qPCR), the locations and expressions of MPO and ICAM-1 proteins were observed by immunohistochemical staining, and the expressions of TGF-β1, Smad3, p-Smad3 and ICAM-1 proteins in lung tissue were detected by Western blot.Results At different time points after irradiation, the pathological manifestations such as inflammation and exudation of lung tissue in the RT+CRZ group were significantly increased, and the total number of cells and protein content in BALF was higher than that of the other three groups, compared with RT group, the difference was statistically significant at 4 week (t=-5.031, -2.814, P < 0.05). Compared with RT group, the expressions of ICAM-1 and TGF-β1 mRNA in lung tissue of the RT+CRZ group were significantly increased, while the expression of TGF-β1 increased significantly at 1, 4 and 8 weeks after irradiation (t=-2.687, -7.032, -5.221, P < 0.05), and the expression of ICAM-1 increased significantly at 2 and 4 weeks after irradiation (t=-4.819, -6.057, P < 0.05).The expressions of these two gradually increased from 1 to 4 weeks and peaked in 4 weeks, then decreased at 8 weeks. At the same time, the trend of the expression of MPO mRNA was consistent with ICAM-1 and TGF-β1. At 4 week, there was no difference in the expression of Smad3 protein in these four groups (P>0.05). The expressions of TGF-β1, p-Smad3, ICAM-1 and p-Smad3/Smad3 proteins of the RT+CRZ group were all higher than those of the other three groups (F=14.74, 10.03, 35.29, 22.94, P < 0.05).Conclusions Crizotinib combined with radiotherapy can aggravate acute radiation-induced lung injury, which may due to the increase of ICAM-1 expression by up-regulating the TGF-β1 signaling pathway. |
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