Inhibition of transforming growth factor-β type I receptor signaling by RNA interference suppresses the proliferation, collagen synthesis, and inflammation factors in human embryonic lung fibroblasts

Shihui Lin^a,†, Juan Fu^b,a,†, Chuanjiang Wang^a, Qiong Liu^a, Fang Xu^a,, Yuanzheng Yang^c,

ABSTRACT: Transforming growth factor-β1 (TGF-β1) is an important mediator of pulmonary fibrosis. The purpose of this study was to determine whether small interference RNA (siRNA) directed to the transforming growth factor-β type I receptor (TβRI) could be used to suppress the action of TGF-β1 in human embryonic lung fibroblasts (HELF). Here we constructed 4 specific recombinant lentivirus vectors, targeting the TβRI gene to transfect the HELF. We observed their effects on TβRI expression using real-time PCR and Western blot. The HELF were stimulated with TGF-β1 after transfection with lentivirus vectors. The expression of p-Smad3 was measured using Western blot. The proliferation activity of the HELF was determined with an MTT assay, the cell cycle was analyzed using flow cytometry, the collagen expression was observed with Sirius Red staining, the expression of α-smooth muscle actin (α-SMA best hunting crossbow) was measured using RT-PCR, and the supernatant tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), collagen I and collagen III levels were measured with ELISA testing. We found that TβRI siRNA abrogated the receptor transcription and protein expression in growing HELF. The lentivirus-mediated siRNA targeting the TβRI gene could inhibit the activation of HELF and the expression of p-Smad3 stimulated by TGF-β1, and effectively suppressed its effects on cell proliferation, the cell cycle, collagen synthesis, α-SMA, collagen I, collagen III, TNF-α and IL-6 expression in HELF. TβRI-specific siRNAs were efficacious in knocking down TGF-β1 action in vitro. Application of siRNA in HELF to downregulate TβRI expression may provide a novel therapy for preventing pulmonary inflammation and fibrosis.

Download PDF

44 Downloads 904 Views

^a	Department of Emergency and Critical Care Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016 China
^b	Department of Emergency, Foshan First Peopl?s Hospital, Foshan 528000 China
^c	Department of Critical Care Medicine, The Affiliated Hospital of Hainan Medical College, Haikou 570102 China

* Corresponding author, E-mail: xufang828@126.com, hhyangyuanzheng@163.com

Received 5 Dec 2019, Accepted 10 Jul 2020

Inhibition of transforming growth factor-β type I receptor signaling by RNA interference suppresses the proliferation, collagen synthesis, and inflammation factors in human embryonic lung fibroblasts

Shihui Lina,†, Juan Fub,a,†, Chuanjiang Wanga, Qiong Liua, Fang Xua,*, Yuanzheng Yangc,*

Shihui Lin^a,†, Juan Fu^b,a,†, Chuanjiang Wang^a, Qiong Liu^a, Fang Xu^a,, Yuanzheng Yang^c,