摘要
目的肺纤维化中过多的成纤维细胞(FBs)活化推动着基质蛋白沉积、纤维化进展。黏着斑蛋白(Vinculin)是一种黏着斑成分蛋白,课题组前期研究发现vinculin与FBs活化有关,但体内改变FBs的vinculin表达量对肺纤维化进程的影响尚不清楚。方法C57BL/6小鼠全肺照射形成放射性肺纤维化(RIPF),静脉注射vinculin敲低细胞及vinculin突变细胞,于照射后4个月取小鼠肺组织,采用Masson染色、血相分析了解肺纤维化病理变化及炎症情况,RT-PCR、Western blot及免疫荧光法了解肺组织中基质蛋白含量、FBs活化等的变化。结果体外实验显示敲低vinculin可下调FBs的增殖、活化及迁移能力,而过表达突变vinculin在体外培养中未见对FBs的显著调节作用。给RIPF小鼠分别注射vinculin敲低细胞和vinculin突变细胞,结果发现注射vinculin敲低细胞可减少胶原蛋白沉积,下调血相炎症指标,下调肺组织中vinculin及α-SMA表达量,而注射vinculin突变细胞对RIPF的部分上述指标具有上调作用。结论本实验结果显示vinculin表达量与RIPF的进展有关,注射vinculin敲低细胞具有一定的缓解RIPF的作用。
OBJECTIVE The excessive activation of fibroblasts(FBs)promotes matrix protein deposition and pulmonary fibrosis.Vinculin is a component of focal adhesion,and the previous study of the tutor group found that vinculin is related to the activation of FBs,but the effect of changing the expression of vinculin in FBs in vivo on the process of pulmonary fibrosis is not clear.METHODS Radiation induced pulmonary fibrosis(RIPF)was established in C57BL/6 by thoracic irradiation.Mice were intravenously injected with vinculin knockdown cells and vinculin mutant cells,the lung tissue was taken 4 months after irradiation,and pathological changes and inflammation of pulmonary fibrosis were analyzed by Masson staining and blood phase analysis,and the changes of matrix protein content and FBs activation in lung tissues were detected by RT-PCR,Western blot and immunofluorescence methods.RESULTS In vitro experiments showed that knocking down vinculin could downregulate the proliferation,activation and migration ability of FBs,while overexpression mutation vinculin did not have a significant regulatory effect on FBs.RIPF mice were injected with vinculin knockdown cells and vinculin mutant cells,and it was found that injecting vinculin knockdown cells could reduce collagen deposition,down-regulate blood cells and the expression of vinculin andα-SMA in lung tissue,while injecting vinculin mutant cells could upregulate some of the above indicators of RIPF.CONCLUSION Vinculin expression is related to the progression of RIFF,and the injection of vinculin knockdown cells has a certain effect on alleviating RIFF.
作者
杨超
郑姝琦
谢灵枫
唐轶韬
吴楚
陈纯
YANG Chao;ZHENG Shu-qi;XIE Ling-feng;TANG Yi-tao;WU Chu;CHEN Chun(019 Clinical Pharmacy Major,Fujian Medical University,Fuzhou 350122,China;2019 Clinical Medicine Major,Fujian Medical University,Fuzhou 350122,China;College of Pharmacy,Fujian Medical University,Fuzhou 350122,China;Fujian Key Laboratory of Natural Medicine Pharmacology,Fuzhou 350122,China)
出处
《海峡药学》
2023年第1期6-12,共7页
Strait Pharmaceutical Journal
基金
福建医科大学“大学生创新创业训练计划项目”(NO:202110392015)。