Fluorofenidone attenuates paraquat‑induced pulmonary fibrosis by regulating the PI3K/Akt/mTOR signaling pathway and autophagy

Jiang,Feiya; Li,Sha; Jiang,Yu; Chen,Zhuo; Wang,Tongtong; Liu,Wen

doi:10.3892/mmr.2021.12044

Fluorofenidone attenuates paraquat‑induced pulmonary fibrosis by regulating the PI3K/Akt/mTOR signaling pathway and autophagy

Authors:
- Feiya Jiang
- Sha Li
- Yu Jiang
- Zhuo Chen
- Tongtong Wang
- Wen Liu
View Affiliations

Affiliations: Department of Pharmacy, The First Hospital Affiliated with Hunan Normal University, Changsha, Hunan 410005, P.R. China, Department of Pharmacy, Changsha Stomatological Hospital, Changsha, Hunan 410004, P.R. China, Department of Emergency, The First Hospital Affiliated with Hunan Normal University, Changsha, Hunan 410005, P.R. China, Department of Pharmacy, Central South University, Changsha, Hunan 410013, P.R. China
Published online on: March 26, 2021 https://doi.org/10.3892/mmr.2021.12044
Article Number: 405
Copyright: © Jiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Paraquat (PQ) is a widely used herbicide that is severely toxic to humans and animals. Pulmonary fibrosis is a disorder that can result from PQ poisoning. Fluorofenidone (AKF‑PD) is a novel small molecule pyridone drug with a widespread and clear anti‑organ fibrosis effect; however, its mechanism of action on PQ poisoning‑induced pulmonary fibrosis is not clear. The purpose of the present study was to investigate the protective effect and underlying mechanism of AKF‑PD on PQ poisoning‑induced pulmonary fibrosis. Human alveolar epithelial cells (HPAEpiC) and Sprague‑Dawley rats were treated with AKF‑PD in the presence or absence of PQ. Hematoxylin‑eosin and Masson staining were used to observe the morphological changes in lung tissue. Cell Counting Kit‑8 and lactate dehydrogenase assays were used to evaluate the viability of HPAEpiC cells. ELISA was used to detect inflammatory factors and the collagen content. Finally, the effects of AKF‑PD on pulmonary fibrosis, as well as the underlying mechanisms, were evaluated via western blotting, reverse transcription‑quantitative PCR and immunofluorescence analysis. AKF‑PD effectively alleviated PQ‑induced pulmonary fibrosis and reduced the expression of oxidative stress and inflammatory factors. Moreover, AKF‑PD treatment effectively inhibited the PI3K/Akt/mTOR signaling pathway and upregulated autophagy. Overall, these findings suggested that AKF‑PD can alleviate PQ‑induced inflammation and pulmonary fibrosis by inhibiting the PI3K/Akt/mTOR signaling pathway and by upregulating autophagy.

View Figures

View References

1	Samsamshariat S, Vedaei A, Jahangiri S, Gavarti MB, Sami R, Taheri A and Dorooshi G: Report of a case of paraquat poisoning and mediastinal involvement. Adv Biomed Res. 10:52021. View Article : Google Scholar
2	Zhu Y, Tan J, Xie H, Wang J, Meng X and Wang R: HIF-1α regulates EMT via the Snail and β-acatenin pathways in paraquat poisoning-induced early pulmonary fibrosis. J Cell Mol Med. 20:688–697. 2016. View Article : Google Scholar : PubMed/NCBI
3	Suntres ZE: Role of antioxidants in paraquat toxicity. Toxicology. 180:65–77. 2002. View Article : Google Scholar : PubMed/NCBI
4	Xiao Q, Wang W, Qi H, Gao X, Zhu B, Li J and Wang P: Continuous hemoperfusion relieves pulmonary fibrosis in patients with acute mild and moderate paraquat poisoning. J Toxicol Sci. 45:611–617. 2020. View Article : Google Scholar : PubMed/NCBI
5	Tao W, Shu Y, Miao Q and Zhu Y: Attenuation of hyperoxia-induced lung injury in rats by adrenomedullin. Inflammation. 35:150–157. 2012. View Article : Google Scholar : PubMed/NCBI
6	Shen H, Wu N, Wang Y, Zhao H, Zhang L, Li T and Zhao M: Chloroquine attenuates paraquat-induced lung injury in mice by altering inflammation, oxidative stress and fibrosis. Int Immunopharmacol. 46:16–22. 2017. View Article : Google Scholar : PubMed/NCBI
7	Bai L, Li A, Gong C, Ning X and Wang Z: Protective effect of rutin against bleomycin induced lung fibrosis: Involvement of TGF-β1/α-SMA/Col I and III pathway. Biofactors. 46:634–644. 2020. View Article : Google Scholar
8	Qian J, Ye Y, Lv L, Zhu C and Ye S: FTY720 attenuates paraquat-induced lung injury in mice. Int Immunopharmacol. 21:426–431. 2014. View Article : Google Scholar : PubMed/NCBI
9	Xu S, Hu H, Jiang Z, Tang S, Zhou Y, Sheng J, Chen J and Cao Y: APACHE score, severity index of paraquat poisoning, and serum lactic acid concentration in the prognosis of paraquat poisoning of Chinese patients. Pediatr Emerg Care. 31:117–121. 2015. View Article : Google Scholar : PubMed/NCBI
10	Yang H, Zhang W, Xie T, Wang X and Ning W: Fluorofenidone inhibits apoptosis of renal tubular epithelial cells in rats with renal interstitial fibrosis. Braz J Med Biol Res. 52:e87722019. View Article : Google Scholar : PubMed/NCBI
11	Song C, He L, Zhang J, Ma H, Yuan X, Hu G, Tao L, Zhang J and Meng J: Fluorofenidone attenuates pulmonary inflammation and fibrosis via inhibiting the activation of NALP3 inflammasome and IL-1β/IL-1R1/MyD88/NF-κB pathway. J Cell Mol Med. 20:2064–2077. 2016. View Article : Google Scholar : PubMed/NCBI
12	Ma H, Peng Z, Hu G and Tao L: Effect and mechanism of fluorofenidone on organ fibrosis. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 40:208–213. 2015.(In Chinese). PubMed/NCBI
13	Meng J, Zou Y, Hu C, Zhu Y, Peng Z, Hu G, Wang Z and Tao L: Fluorofenidone attenuates bleomycin-induced pulmonary inflammation and fibrosis in mice via restoring caveolin 1 expression and inhibiting mitogen-activated protein kinase signaling pathway. Shock. 38:567–573. 2012. View Article : Google Scholar : PubMed/NCBI
14	Tang J, Li J, Li G, Zhang H, Wang L, Li D and Ding J: Spermidine-mediated poly(lactic-co-glycolic acid) nanoparticles containing fluorofenidone for the treatment of idiopathic pulmonary fibrosis. Int J Nanomedicine. 12:6687–6704. 2017. View Article : Google Scholar : PubMed/NCBI
15	Xu M, Li S, Wang J, Huang S, Zhang A, Zhang Y, Gu W, Yu X and Jia Z: Cilomilast ameliorates renal tubulointerstitial fibrosis by inhibiting the TGF-β1-Smad2/3 signaling pathway. Front Med (Lausanne). 7:6261402021. View Article : Google Scholar : PubMed/NCBI
16	Liu J, Song C, Xiao Q, Hu G, Tao L and Meng J: Fluorofenidone attenuates TGF-β1-induced lung fibroblast activation via restoring the expression of caveolin-1. Shock. 43:201–207. 2015. View Article : Google Scholar : PubMed/NCBI
17	Saxton RA and Sabatini DM: mTOR signaling in growth, metabolism, and disease. Cell. 168:960–976. 2017. View Article : Google Scholar : PubMed/NCBI
18	Alayev A and Holz MK: mTOR signaling for biological control and cancer. J Cell Physiol. 228:1658–1664. 2013. View Article : Google Scholar : PubMed/NCBI
19	Gui YS, Wang L, Tian X, Xue L, Ma A, Zhou W, Zeng N, Ji Z, Cai B, Zhang H, et al: mTOR overactivation and compromised autophagy in the pathogenesis of pulmonary fibrosis. PLoS One. 10:e01386252015. View Article : Google Scholar : PubMed/NCBI
20	Gui X, Chen H, Cai H, Sun L and Gu L: Leptin promotes pulmonary fibrosis development by inhibiting autophagy via PI3K/Akt/mTOR pathway. Biochem Biophys Res Commun. 498:660–666. 2018. View Article : Google Scholar : PubMed/NCBI
21	Rieg AD, Said S, Carolin A, Eva V, Rolf R, Stefan U and Christian M: PDGF-BB regulates the pulmonary vascular tone: Impact of prostaglandins, calcium, MAPK- and PI3K/AKT/mTOR signalling and actin polymerisation in pulmonary veins of guinea pigs. Respir Res. 19:1202018. View Article : Google Scholar : PubMed/NCBI
22	Szapiel SV, Elson NA, Fulmer JD, Hunninghake GW and Crystal RG: Bleomycin-induced interstitial pulmonary disease in the nude, athymic mouse. Am Rev Respir Dis. 120:893–899. 1979.PubMed/NCBI
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
24	Liu MW, Su MX, Tang DY, Hao L, Xun XH and Huang YQ: Ligustrazin increases lung cell autophagy and ameliorates paraquat-induced pulmonary fibrosis by inhibiting PI3K/Akt/mTOR and hedgehog signalling via increasing miR-193a expression. BMC Pulm Med. 19:352019. View Article : Google Scholar : PubMed/NCBI
25	Raghu G, Rochwerg B, Zhang Y, Garcia CA, Azuma A, Behr J, Brozek JL, Collard HR, Cunningham W, Homma S, et al: An official ATS/ERS/JRS/ALAT clinical practice guideline: Treatment of idiopathic pulmonary fibrosis. An update of the 2011 clinical practice guideline. Am J Respir Crit Care Med. 192:e3–19. 2015. View Article : Google Scholar : PubMed/NCBI
26	Wang J, Qiao L, Li S and Yang G: Protective effect of ginsenoside Rb1 against lung injury induced by intestinal ischemia-reperfusion in rats. Molecules. 18:1214–1226. 2013. View Article : Google Scholar : PubMed/NCBI
27	Wu J, Chu Z, Ruan Z, Wang X, Dai T and Hu X: Changes of intracellular porphyrin, reactive oxygen species, and fatty acids profiles during inactivation of methicillin-resistant staphylococcus aureus by antimicrobial blue light. Front Physiol. 9:16582018. View Article : Google Scholar : PubMed/NCBI
28	Yamada M, Kuwano K, Maeyama T, Hamada N Michihiro Yoshimi, Yoichi Nakanishi and Kasper M: Dual-immunohistochemistry provides little evidence for epithelial-mesenchymal transition in pulmonary fibrosis. Histochem Cell Biol. 129:453–462. 2008. View Article : Google Scholar : PubMed/NCBI
29	Molina-Molina M, Machahua-Huamani C, Vicens-Zygmunt V, Llatjós R, Escobar I, Sala-Llinas E, Luburich-Hernaiz P, Dorca J and Montes-Worboys A: Anti-fibrotic effects of pirfenidone and rapamycin in primary IPF fibroblasts and human alveolar epithelial cells. BMC Pulm Med. 18:632018. View Article : Google Scholar : PubMed/NCBI
30	Chitra P, Saiprasad G, Manikandan R and Sudhandiran G: Berberine inhibits Smad and non-Smad signaling cascades and enhances autophagy against pulmonary fibrosis. J Mol Med (Berl). 93:1015–1031. 2015. View Article : Google Scholar : PubMed/NCBI
31	Liu Z, Li Y, Song H, He J, Li G, Zheng Y and Li B: Collagen peptides promote photoaging skin cell repair by activating the TGF-β/Smad pathway and depressing collagen degradation. Food Funct. 10:6121–6134. 2019. View Article : Google Scholar : PubMed/NCBI
32	Eskelinen E: Autophagy: Supporting cellular and organismal homeostasis by self-eating. Int J Biochem Cell Biol. 111:1–10. 2019. View Article : Google Scholar : PubMed/NCBI
33	Lv X, Li K and Hu Z: Autophagy and Pulmonary Fibrosis. Adv Exp Med Biol:. 1207:569–579. 2020. View Article : Google Scholar : PubMed/NCBI
34	Tian K, Chen P, Liu Z, Si S, Zhang Q, Mou Y, Han L, Wang Q and Zhou X: Sirtuin 6 inhibits epithelial to mesenchymal transition during idiopathic pulmonary fibrosis via inactivating TGF-β1/Smad3 signaling. Oncotarget. 8:61011–61024. 2017. View Article : Google Scholar : PubMed/NCBI
35	Mu E, Wang J, Chen L, Lin S, Chen J and Huang S: Ketogenic diet induces autophagy to alleviate bleomycin-induced pulmonary fibrosis in murine models. Exp Lung Res. 47:26–36. 2021. View Article : Google Scholar : PubMed/NCBI
36	Im J, Hergert P and Nho R: Reduced FoxO3a expression causes low autophagy in idiopathic pulmonary fibrosis fibroblasts on collagen matrices. Am J Physiol Lung Cell Mol Physiol. 309:L552–L561. 2015. View Article : Google Scholar : PubMed/NCBI
37	Xie T, Xu Q, Wan H, Xing S, Shang C, Gao Y and He Z: Lipopolysaccharide promotes lung fibroblast proliferation through autophagy inhibition via activation of the PI3K-Akt-mTOR pathway. Lab Invest. 99:625–633. 2019. View Article : Google Scholar : PubMed/NCBI
38	Patel AS, Lin L, Geyer A, Haspel JA, An CH, Cao J, Rosas IO and Morse D: Autophagy in idiopathic pulmonary fibrosis. PLoS One. 3:e413942012. View Article : Google Scholar
39	Jung C, Ro S, Cao J, Otto N and Kim D: mTOR regulation of autophagy. FEBS Lett. 584:1287–1295. 2010. View Article : Google Scholar : PubMed/NCBI
40	Nho R and Hergert P: IPF fibroblasts are desensitized to type I collagen matrix-induced cell death by suppressing low autophagy via aberrant Akt/mTOR kinases. PLoS One. 9:e946162014. View Article : Google Scholar : PubMed/NCBI