[1]黄玉,王子文,王会兰,等.七甲川花菁类荧光小分子IR-61改善大鼠肺纤维化[J].第三军医大学学报,2020,42(04):350-358.
 HUANG Yu,WANG Ziwen,WANG Huilan,et al.Fluorescent small molecule IR-61 improves pulmonary fibrosis induced by intratracheal bleomycin in rats[J].J Third Mil Med Univ,2020,42(04):350-358.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第04期
页码:
350-358
栏目:
基础医学
出版日期:
2020-02-29

文章信息/Info

Title:
Fluorescent small molecule IR-61 improves pulmonary fibrosis induced by intratracheal bleomycin in rats
作者:
黄玉王子文王会兰罗敏张爱华史春梦
贵州医科大学公共卫生学院,环境污染与疾病监控教育部重点实验室; 陆军军医大学(第三军医大学)军事预防医学系火箭军医学教研室,创伤、烧伤与复合伤国家重点实验室;西南医科大学附属医院肿瘤科
Author(s):
HUANG Yu12 WANG Ziwen2 WANG Huilan3 LUO Min12 ZHANG Aihua1 SHI Chunmeng12

1Key Laboratory of Environmental Pollution Monitoring and Disease Control of Ministry of Education, School of Public Health, Guizhou Medical University, Guiyang, Guizhou Province, 550025; 2State Key Laboratory of Trauma, Burns and Combined Injury, Department of Rocket Force Medicine, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038;3Department of Oncology, Affiliated Hospital of  Southwest Medical University, Luzhou, Sichuan Province, 646000, China

关键词:
IR-61肺纤维化成纤维细胞细胞转分化细胞增殖细胞迁移
Keywords:
IR-61 pulmonary fibrosis lung fibroblasts cell transdifferentiation cell proliferation cell migration
分类号:
R563.13;R966;R977.6
文献标志码:
A
摘要:

目的 探索七甲川花菁类荧光小分子IR-61在肺纤维化发生过程中的作用以及对肺成纤维细胞转分化、增殖和迁移的影响。方法 采用气管滴注博来霉素(bleomycin, BLM,2.5 mg/kg)建立大鼠肺纤维化模型;腹腔注射PBS或IR-61(2.0 mg/kg,每周2次);BLM处理10 d后,尾静脉注射IR-61(2.0 mg/kg),于24 h后取各器官行近红外荧光成像(near infrared fluorescence imaging,NIR),观察IR-61在各器官的蓄积情况;治疗28 d后,取肺组织制作切片,行HE和Masson染色,显微镜下观察;采用RT-PCR、Western blot测定人肺成纤维细胞(human fetal lung fibroblast, HFL1)与原代大鼠肺成纤维细胞转分化及增殖相关基因Collagen-1、Collagen-3、α-SMA、Fibronectin、FOXM1、CCNB1、CDC25b和AURKB的mRNA及蛋白表达;用细胞划痕实验进行迁移检测。结果 ①NIR显示:IR61可靶向至博来霉素诱导的受损伤肺组织。②形态学观察显示:BLM+IR61组较博来霉素组形态改善,肺塌陷少,纤维结节少。经组织学鉴定,BLM+IR61处理组较BLM处理组肺纤维化程度低[(0.103 3±0.0145 3)vs(0.460 0±0.0378 6),P<0.05] 。IR-61预处理HFL1与原代大鼠肺成纤维细胞后,用TGFβ1刺激,Western blot和RTPCR分析结果显示:TGF-β1+IR-61处理组较TGFβ1处理组转分化特异性基因Collagen-1、Collagen-3、α-SMA、Fibronectin 的mRNA及蛋白表达降低(P<0.05),同时,增殖特异性FOXM1、CCNB1、CDC25b和AURKB基因的表达也降低(P<0.05),说明IR-61可以抑制TGF-β1诱导的肺成纤维细胞的转分化与增殖的作用。③细胞划痕实验结果显示:TGF-β1诱导肺成纤维细胞迁移活性增强,IR-61能有效抑制肺成纤维细胞的迁移能力。结论 IR-61通过靶向到受损伤的肺组织并通过抑制肺成纤维细胞的转分化、增殖和迁移来减轻博来霉素诱导的肺纤维化。

Abstract:

Objective To evaluate the effects of IR61 on the development of pulmonary fibrosis and on the proliferation and migration of lung fibroblasts in rats.  Methods Forty male SD rats (6 to 8 weeks old) were randomized equally into normal control group, IR-61 group, bleomycin group and bleomycin+IR61 group. In the latter 2 groups, rat models of pulmonary fibrosis were established by intratracheal infusion of bleomycin (2.5 mg/kg). In the 2 groups with IR-61 treatment, IR-61 was intraperitoneally injected at the dose of 2.0 mg/kg 30 min following intratracheal bleomycin infusion and was then administered twice a week. At 10 d after bleomycin treatment, 3 rats from each group were selected for IR-61 injection (2 mg/kg) via the tail vein to observe the accumulation of IR-61 in each organ at 24 h using near infrared fluorescence imaging (NIR). After 28 d of treatment, the rats were euthanized and the lung tissue was collected for HE and Masson staining. RT-PCR and Western blotting were performed to detect the changes of the expression of the genes related with lung fibroblast transdifferentiation and proliferation (including Collagen-1, Collagen-3,α-SMA, fibronectin, FOXM1, CCNB1, CDC25b and AURKB) in human lung fibroblasts (HFL1) and primary rat lung fibroblasts in response to IR-61 and transforming growth factor-β1 (TGF-β1) treatments; the migration of the cells was assessed using cell scratch test. Results imaging showed that IR-61 could target the injured lung tissue induced by bleomycin. Morphological observation showed that compared with bleomycintreated rats, the rats with IR-61 treatment presented with better structural integrity of the lungs with less obvious lung collapse, fewer fibrotic nodules and a significantly milder of pulmonary fibrosis (0.103 3±0.014 53 vs0.460 0±0.037 86, P<0.05). In HFL1 cells and primary rat lung fibroblasts, IR-61 pretreatment prior to TGF-β1 stimulation resulted in significantly lowered mRNA and protein expression of collagen-1, collagen-3, α-MA, and fibronectin as well as the proliferationspecific genes FOXM1, CCNB1, CDC25b and AURKB (P<0.05). Cell scratch test showed that TGF-β1 treatment significantly increased the cell migration activity, which was effectively suppressed by IR-61 treatment. Conclusion61 ameliorates bleomycininduced pulmonary fibrosis in rats by targeting the injured lung tissue and inhibiting the transdifferentiation, proliferation and migration of lung fibroblasts.

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更新日期/Last Update: 2020-02-24