Jang Jun-Ho, Chand Hitendra S, Bruse Shannon, Doyle-Eisele Melanie, Royer Christopher, McDonald Jacob, Qualls Clifford, Klingelhutz Aloysius J, Lin Yong, Mallampalli Rama, Tesfaigzi Yohannes, Nyunoya Toru
a Department of Medicine , University of Pittsburgh , Pittsburgh , PA , USA.
b VA Pittsburgh Healthcare System , Pittsburgh , PA , USA.
COPD. 2017 Apr;14(2):228-237. doi: 10.1080/15412555.2016.1262340. Epub 2016 Dec 27.
The purpose of this study was to determine whether expression of connective tissue growth factor (CTGF) protein in chronic obstructive pulmonary disease (COPD) is consistent in humans and animal models of COPD and to investigate the role of this protein in lung epithelial cells. CTGF in lung epithelial cells of ex-smokers with COPD was compared with ex-smokers without COPD by immunofluorescence. A total of twenty C57Bl/6 mice and sixteen non-human primates (NHPs) were exposed to cigarette smoke (CS) for 4 weeks. Ten mice of these CS-exposed mice and eight of the CS-exposed NHPs were infected with H3N2 influenza A virus (IAV), while the remaining ten mice and eight NHPs were mock-infected with vehicle as control. Both mRNA and protein expression of CTGF in lung epithelial cells of mice and NHPs were determined. The effects of CTGF overexpression on cell proliferation, p16 protein, and senescence-associated β-galactosidase (SA-β-gal) activity were examined in cultured human bronchial epithelial cells (HBECs). In humans, CTGF expression increased with increasing COPD severity. We found that protein expression of CTGF was upregulated in lung epithelial cells in both mice and NHPs exposed to CS and infected with IAV compared to those exposed to CS only. When overexpressed in HBECs, CTGF accelerated cellular senescence accompanied by p16 accumulation. Both CTGF and p16 protein expression in lung epithelia are positively associated with the severity of COPD in ex-smokers. These findings show that CTGF is consistently expressed in epithelial cells of COPD lungs. By accelerating lung epithelial senescence, CTGF may block regeneration relative to epithelial cell loss and lead to emphysema.
本研究的目的是确定慢性阻塞性肺疾病(COPD)中结缔组织生长因子(CTGF)蛋白的表达在人类和COPD动物模型中是否一致,并研究该蛋白在肺上皮细胞中的作用。通过免疫荧光比较了患有COPD的前吸烟者与未患COPD的前吸烟者肺上皮细胞中的CTGF。总共20只C57Bl/6小鼠和16只非人类灵长类动物(NHP)暴露于香烟烟雾(CS)中4周。这些暴露于CS的小鼠中有10只和暴露于CS的NHP中有8只感染了H3N2甲型流感病毒(IAV),而其余10只小鼠和8只NHP则用载体进行假感染作为对照。测定了小鼠和NHP肺上皮细胞中CTGF的mRNA和蛋白表达。在培养的人支气管上皮细胞(HBEC)中检测了CTGF过表达对细胞增殖、p16蛋白和衰老相关β-半乳糖苷酶(SA-β-gal)活性的影响。在人类中,CTGF表达随着COPD严重程度的增加而增加。我们发现,与仅暴露于CS的小鼠和NHP相比,暴露于CS并感染IAV的小鼠和NHP肺上皮细胞中CTGF的蛋白表达上调。当在HBEC中过表达时,CTGF加速细胞衰老并伴有p16积累。肺上皮细胞中CTGF和p16蛋白表达均与前吸烟者COPD的严重程度呈正相关。这些发现表明,CTGF在COPD肺上皮细胞中持续表达。通过加速肺上皮衰老,CTGF可能会阻止相对于上皮细胞丢失的再生并导致肺气肿。