Transforming growth factor-β1/Smads signaling pathway involves the development of paraquat-induced pulmonary endothlial-mesenchymal transition and fibrosis

doi:10.3877/cma.j.issn.1674-6880.2018.01.001

Abstract

Abstract:

Objective

To observe the endothlial-mesenchymal transition (EMT) phenomenon and related protein expressions in transforming growth factor-β1 (TGF-β1)/Smads signaling pathway in lung tissues of the rats with paraquat-induced pulmonary fibrosis.

Methods

Sixty male SD rats were randomly divided into paraquat group (n=30) and control group (n=30). The paraquat poisoning model was established by intraperitoneally injecting with 80 mg/kg of paraquat saline solution in the paraquat group, while rats in the control group were injected with 3 mL of sterile saline. The rats were sacrificed in batches at 1, 3, 7, 14, 21, 28 d after modeling. The wet/dry ratio of left lung tissue and the expression of hydroxyproline were calculated between these two groups. The pathology and pulmonary fibrosis in the inferior lobe of left lung were assessed by hematoxylin-eosin (HE) staining and Masson staining. The expressions of related proteins in EMT and TGF-β1/Smads signaling pathway in lung tissues on days 1, 3, 7, 14, 21, 28 after paraquat poisoning were analyzed by Western-blotting.

Results

The wet/dry ratio and hydroxyproline both showed significant differences between two groups at different time points after modeling (F=9.772, 22.541, both P < 0.001). In the paraquat group, the wet/dry ratios at 3, 7, 14, 21, 28 d were much higher than those in the control group (all P<0.05), and the levels of hydroxyproline only increased at 14, 21, 28 d as compared with those in the control group (all P<0.05). Pathological findings indicated that inflammatory cell infiltrated, alveolar interstitium thickened in the paraquat group at 1, 3, 7 d, and alveolar walls thickened gradually and a large amount of collagen fibers proliferated at 21, 28 d after modeling. There was no significant change in lung tissue of the control group. The expressions of TGF-β1, Smad2/3, p-Smad2/3, Smad4, p-Smad4, Smad7, p-Smad7, E-cadherin (E-cad), α-smooth muscle actin (α-SMA), Vimentin and fibroblast specific protein-1 (FSP-1) between the two groups at each time point all showed significant differences (F=10.685, 7.381, 7.878, 10.743, 14.575, 17.791, 33.200, 14.453, 10.849, 25.415, 26.263, all P < 0.001). The expressions of TGF-β1, Smad2/3, p-Smad2/3, Smad4, p-Smad4, α-SMA, Vimentin and FSP-1 gradually increased and reached a peak at 21 d after paraquat poisoning, while the Smad7, p-Smad7 decreased and reached a bottom at 7 d in the paraquat group (all P<0.05). The expression of E-cad in the paraquat group reduced from 3 d to 21 d (all P<0.05).

Conclusion

The TGF-β1/Smads signaling pathway may affect the development of PQ-induced pulmonary fibrosis by regulating EMT process.

Key words: Transforming growth factor beta1, Paraquat, Poisoning, Pulmonary fibrosis, Epithelial-mesenchymal transition, Rats

Fengxuan Wu, Jingchen Zhang, Jiukun Jiang, Yuanqiang Lu. Transforming growth factor-β1/Smads signaling pathway involves the development of paraquat-induced pulmonary endothlial-mesenchymal transition and fibrosis[J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2018, 11(01): 3-10.

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组别	只数	1d	3d	7 d	14 d	21 d	28 d
对照组	30	3.8 ± 0.3	4.0 ± 0.5	3.9 ± 0.4	4.2 ± 0.4	4.1 ± 0.7	3.9 ± 0.8
百草枯组	30	4.2 ± 0.9	5.9 ± 1.1^ab	6.2 ± 1.3^ab	6.8 ± 1.4^ab	7.3 ± 1.6^ab	7.1 ± 1.5^ab

组别	只数	1d	3d	7 d	14 d	21 d	28 d
对照组	30	3.8 ± 0.3	4.0 ± 0.5	3.9 ± 0.4	4.2 ± 0.4	4.1 ± 0.7	3.9 ± 0.8
百草枯组	30	4.2 ± 0.9	5.9 ± 1.1^ab	6.2 ± 1.3^ab	6.8 ± 1.4^ab	7.3 ± 1.6^ab	7.1 ± 1.5^ab

组别	只数	1d	3d	7 d	14 d	21 d	28 d
对照组	30	2.47 ± 0.83	3.31 ± 0.72	2.80 ± 0.63	2.52 ± 0.73	3.21 ± 1.14	2.63 ± 1.24
百草枯组	30	2.73 ± 1.23	4.63 ± 0.85	5.24 ± 1.64	7.33 ± 1.33^ab	8.34 ± 0.97^ab	9.68 ± 1.22^ab

组别	只数	1d	3d	7 d	14 d	21 d	28 d
对照组	30	2.47 ± 0.83	3.31 ± 0.72	2.80 ± 0.63	2.52 ± 0.73	3.21 ± 1.14	2.63 ± 1.24
百草枯组	30	2.73 ± 1.23	4.63 ± 0.85	5.24 ± 1.64	7.33 ± 1.33^ab	8.34 ± 0.97^ab	9.68 ± 1.22^ab

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