Biochemical and Biophysical Research Communications. 2020 Aug 24
Gen-Cheng Gong, Sheng-RenSonga, XinXu, QunLuo, QianHan, Jian-XingHe, JinSu
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Research Field: lung

Keywords: Pulmonary fibrosis, Serpina3n, Chymotrypsin C, Genetic therapy

AAV Serotype: AAV9

Dose: Intratracheal delivery of AAV9 was performed according to the following protocol: 8- to 10-weeks-old mice were injected intraperitoneally and anesthetized suitably with 1% pentobarbital sodium, a catheter was inserted to the trachea, and AAV diluted with 60 mL PBS was pipetted to the top of the catheter to allow animal to gradually breathe in the solution. A titer of 2×10^11 v. g was
administered to every mice, normal group was operated with 60 ml PBS. Three weeks later, AAV-Control and AAV-Serpina3n-KD groups were induced to pulmonary fibrosis by bleomycin, and normal group was operated with the same amount of normal saline.

Routes of Administration: Intratracheal delivery

Targeted organ: lung

Animal or cell line strain: Specific-pathogen free C57BL/6J male mice (8-10 weeks)


Objective: Pulmonary fibrosis is a fatal interstitial lung disease that is characterized by excessive accumulation of extracellular matrix (ECM) and remodeling of lung. The precise mechanisms underlying pulmonary fibrosis still remain unclear. In the current study, we aimed to investigate the alteration and function of serine (or cysteine) peptidase inhibitor, clade A, member 3 N (Serpina3n) in pulmonary fibrotic models and explore the potential mechanisms.

Methods: We induced pulmonary fibrosis in mice by silica and bleomycin respectively and determined Serpina3n in lung tissues, and then verified the expression of Serpina3n and its correlation with pulmonary fibrosis at seven time points in a bleomycin longstanding model. Moreover, adeno-associated virus type 9 (AAV9)-mediated Serpina3n knockdown was used to treat pulmonary fibrosis in the bleomycin model, whose possible mechanisms would be preliminarily explored by detecting chymotrypsin C as an example.

Results: Serpina3n was up-regulated significantly in lungs of both models at mRNA and protein levels relative to control. Notably, the expression of Serpina3n peaked during the 3rd week and then decreased until nearly normal levels during the 10th week, which was closely related to fibrotic procession in bleomycin-treated mice. AAV-mediated Serpina3n knockdown in the lung tissues alleviated bleomycin-induced fibrotic symptoms at various levels and disinhibit chymotrypsin C.

Conclusions: Our study revealed that Serpina3n is a critical regulator in pulmonary fibrosis and suggested Serpina3n inhibition as a potential therapeutic strategy in chronic pulmonary injuries.

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