Effect of hydroxychloroquine sulphate on paraquat-induced lung fibrosis in mice <i>via</i> PI3K/AKt/mTOR signalling pathway_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

DENG Yan , ZHAO Hongyu , ZHU Liping ,  WANG Rongli

Department of Respiratory and Critical Care Medicine, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, P. R. China;

Corresponding author：

WANG Rongli, Email: Scybwrl@sina.com

Keywords：

Pulmonary fibrosis; hydroxychloroquine sulfate; paraquat; PI3K/AKt/mTOR signaling pathway

DOI：

10.7507/1671-6205.202310043

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the effects and mechanisms of hydroxychloroquine sulfate (HCQ) on pulmonary fibrosis through the PI3K/AKt/mTOR signalling pathway. Methods Paraquat intraperitoneal injection was used to establish a mouse model of pulmonary fibrosis. Thirty-six SPF C57BL/6J female mice were randomly divided into a blank group, a paraquat group (20 mg/kg) and a HCQ intervention group. The HCQ intervention group was divided into two subgroups (10 mg/kg and 30 mg/kg) according to different doses. The general condition and body weight changes of mice were observed. twenty-one days later, lung tissues were stained with hematoxylin-eosin and Masson’s pathological staining, and the content of inflammatory factors (IL-1β, IL-6, TNF-α) and hydroxyproline (HYP) were detected by ELISA. Alpha-smooth muscle actin (α-SMA), E-cadherin (E-cad), the expression levels of PI3K/Akt/mTOR pathway-related proteins, phosphatidylinositol 3 kinase (PI3K) and protein kinase B (AKt), and mammalian target of rapamycin (mTOR) were detected by Western blot. The gene expression levels of α-SMA and E-cad were detected by q-PCR. Results Compared with the blank group, the mice in the paraquat group had lower body weight, worse general condition, higher serum levels of inflammatory factors, increased lung structure destruction and collagen deposition, significantly increased HYP content, and higher expression level of PI3K/AKt/mTOR signaling pathway related proteins (all P<0.05). The expression levels of E-cad protein and gene decreased, α-SMA protein and gene increased (all P<0.05). While the HCQ intervention group improved the degree of pulmonary fibrosis in different degrees, and the relevant indexes of PI3K/AKt/mTOR signaling pathway decreased compared with the paraquat group (all P<0.05). Conclusion HCQ can ameliorate paraquat-induced pulmonary fibrosis by inhibiting the PI3K/AKt/mTOR signaling pathway.

Citation： DENG Yan, ZHAO Hongyu, ZHU Liping, WANG Rongli. Effect of hydroxychloroquine sulphate on paraquat-induced lung fibrosis in mice via PI3K/AKt/mTOR signalling pathway. Chinese Journal of Respiratory and Critical Care Medicine, 2024, 23(3): 192-199. doi: 10.7507/1671-6205.202310043 Copy

1.	高利红, 周满红. 百草枯中毒致肺纤维化信号通路研究进展. 中华危重病急救医学, 2021, 33(3): 337-380.
2.	Wang J, Zhu Y, Tan J, et al. Lysyl oxidase promotes epithelial-to-mesenchymal transition during paraquat-induced pulmonary fibrosis. Mol Biosyst, 2016, 12(2): 499-507.
3.	王涛, 刘继才, 逄瑜. 羟氯喹的药理学作用机制及安全性风险分析. 中国药物警戒, 2020, 17(07): 385-388.
4.	Ramser B, Kokot A, Metze D, et al. Hydroxychloroquine modulates metabolic activity and proliferation and induces autophagic cell death of human dermal fibroblasts. J Invest Dermatol, 2009, 129(10): 2419-2426.
5.	Braun S, Ferner M, Kronfeld K, et al. Hydroxychloroquine in children with interstitial (diffuse parenchymal) lung diseases. Pediatr Pulmonol, 2015, 50(4): 410-419.
6.	王超超, 林永强, 蔡小巧, 等. 羟氯喹对IgA肾病患者血清及尿IL-6、TNF-α水平的影响研究. 浙江医学, 2023, 45(5): 514-516, 522.
7.	Li D, Yu K, Feng F, et al. Hydroxychloroquine alleviates renal interstitial fibrosis by inhibiting the PI3K/Akt signaling pathway. Biochem Biophys Res Commun, 2022, 610: 154-161.
8.	马爱平, 高云周, 兰文斌, 等. PI3K/Akt/HIF-1α信号通路在博来霉素诱导小鼠肺纤维化中的作用机制研究. 中国呼吸与危重监护杂志, 2016, 15(1): 34-38.
9.	Lu Y, Zhong W, Liu Y, et al. Anti-PD-L1 antibody alleviates pulmonary fibrosis by inducing autophagy via inhibition of the PI3K/Akt/mTOR pathway. Int Immunopharmacol, 2022, 104: 108504.
10.	王萍, 李铁刚. 百草枯中毒致小鼠肺纤维化模型的建立. 实用药物与临床, 2020, 23(10): 886-889.
11.	杨伟源, 李理, 郑林鑫, 等. EGFR/Foxo3a/Snail1通路在博来霉素肺纤维化小鼠中的作用机制探讨. 中国呼吸与危重监护杂志, 2020, 19(4): 371-378.
12.	Mikawa K, Nishina K, Takao Y, et al. ONO-1714, a nitric oxide synthase inhibitor, attenuates endotoxin-induced acute lung injury in rabbits. Anesth Analg, 2003, 97(6): 1751-1755.
13.	黄海涛, 方静, 曾春晖, 等. 双氢杨梅树皮素对博来霉素诱导小鼠肺纤维化的影响. 中国呼吸与危重监护杂志, 2020, 19(3): 270-275.
14.	刘云华, 周正, 李昌泽, 等. 百草枯中毒致肺纤维化的机制及治疗进展. 毒理学杂志, 2020, 34(2): 166-169,175.
15.	张乙, 陈慧, 崔晋峰. 百草枯诱导肺纤维化机制及治疗研究进展. 临床荟萃, 2019, 34(12): 1134-1137.
16.	李奇, 钱进, 黄琪锋, 等. 百草枯中毒致肺损伤分子机制的研究进展. 海南医学院学报, 2022, 28(4): 309-314, 320.
17.	林琳, 吴开慧, 王文梅. 氯喹/羟氯喹的作用机制及其在口腔疾病中的应用. 口腔疾病防治, 2021, 29(3): 198-201.
18.	崔诚, 么雪婷, 涂思琪, 等. 硫酸羟氯喹的临床药理学研究进展. 中国临床药理学与治疗学, 2020, 25(2): 221-226.
19.	Zheng H, Zhang Y, He J, et al. Hydroxychloroquine inhibits macrophage activation and attenuates renal fibrosis after ischemia-reperfusion injury. Front Immunol, 2021, 12: 645100.
20.	Ji D, Zhao Q, Qin Y, et al. Germacrone improves liver fibrosis by regulating the PI3K/AKT/mTOR signalling pathway. Cell Biol Int, 2021, 45(9): 1866-1875.
21.	Shen H, Wu N, Wang Y, et al. Chloroquine attenuates paraquat-induced lung injury in mice by altering inflammation, oxidative stress and fibrosis. Int Immunopharmacol, 2017, 46: 16-22.
22.	Liu L, Ren J, He Z, et al. Cholesterol-modified hydroxychloroquine-loaded nanocarriers in bleomycin-induced pulmonary fibrosis. Sci Rep, 2017, 7(1): 10737.
23.	郑鑫, 纵加强, 王佳新, 等. 基于Wnt/β-catenin信号通路探讨羟氯喹对烟雾吸入致肺纤维化大鼠的作用机制[EB/OL]. 解放军医学院学报, 1-9. (2023-10-23). http://118.89.52.29:8085/kcms/detail/10.1117.R.20231023.0906.002.html.
24.	Jiang F, Li S, Jiang Y, et al. Fluorofenidone attenuates paraquat-induced pulmonary fibrosis by regulating the PI3K/Akt/mTOR signaling pathway and autophagy. Mol Med Rep, 2021, 23(6): 405.
25.	周佳宁, 肖倩, 黎芳, 等. 基于PI3K/AKT/mTOR信号通路研究瑶药紫九牛醇提物抗小鼠肝纤维化作用. 中药材, 2022, 45(6): 1470-1474.
26.	潘澎, 刘绍能. PI3K/Akt信号通路与肝纤维化. 临床肝胆病杂志, 2013, 29(5): 389-392, 396.
27.	刘珊珊, 盛春瑞, 杜庆红, 等. 麦门冬汤加减方对特发性肺纤维化小鼠PI3K/AKT/mTOR通路的调控. 现代生物医学进展, 2022, 22(17): 3214-3219, 3224.
28.	Hu X, Xu Q, Wan H, et al. PI3K-Akt-mTOR/PFKFB3 pathway mediated lung fibroblast aerobic glycolysis and collagen synthesis in lipopolysaccharide-induced pulmonary fibrosis. Lab Invest, 2020, 100(6): 801-811.
29.	Peng J, Xiao X, Li S, et al. Aspirin alleviates pulmonary fibrosis through PI3K/AKT/mTOR-mediated autophagy pathway. Exp Gerontol, 2023, 172: 112085.
30.	林艺凯, 马爱平, 周伟跃, 等. mTOR信号通路在博来霉素诱导小鼠肺纤维化中的作用机制研究. 中国呼吸与危重监护杂志, 2018, 17(2): 178-182.
31.	Li X, Ma X, Miao Y, et al. Duvelisib attenuates bleomycin-induced pulmonary fibrosis via inhibiting the PI3K/Akt/mTOR signalling pathway. J Cell Mol Med, 2023, 27(3): 422-434.
32.	Qi F, Lv ZD, Huang WD, et al. LncRNA TUG1 promotes pulmonary fibrosis progression via up-regulating CDC27 and activating PI3K/Akt/mTOR pathway. Epigenetics, 2023, 18(1): 2195305.
33.	Hill C, Li J, Liu D, et al. Autophagy inhibition-mediated epithelial-mesenchymal transition augments local myofibroblast differentiation in pulmonary fibrosis. Cell Death Dis, 2019, 10(8): 591.
34.	He J, Peng H, Wang M, et al. Isoliquiritigenin inhibits TGF-β1-induced fibrogenesis through activating autophagy via PI3K/AKT/mTOR pathway in MRC-5 cells. Acta Biochim Biophys Sin (Shanghai), 2020, 52(8): 810-820.
35.	Liu MW, Su MX, Tang DY, et al. 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, 2019, 19(1): 35.
36.	Lu J, Qian Y, Jin W, et al. Hypoxia-inducible factor-1α regulates epithelial-to-mesenchymal transition in paraquat-induced pulmonary fibrosis by activating lysyl oxidase. Exp Ther Med, 2018, 15(3): 2287-2294.
37.	Zhu Y, Tan J, Xie H, et al. HIF-1α regulates EMT via the Snail and β-catenin pathways in paraquat poisoning-induced early pulmonary fibrosis. J Cell Mol Med, 2016, 20(4): 688-697.
38.	Niu Z, Lin J, Hao C, et al. Glycyrrhizic acid attenuates pulmonary fibrosis of silicosis by inhibiting the interaction between HMGB1 and BRG1 through PI3K/Akt/mTOR pathway. Int J Environ Res Public Health, 2022, 19(14): 8743.
39.	Wang Y, Men M, Xie B, et al. Inhibition of PKR protects against H2O2-induced injury on neonatal cardiac myocytes by attenuating apoptosis and inflammation. Sci Rep, 2016, 6: 38753.
40.	何文涓, 袁志坚, 黄寅, 等. 芹菜素对百草枯致小鼠肺纤维化保护作用及机制研究. 药物评价研究, 2021, 44(8): 1660-1665.
41.	Hossain MM, Sonsalla PK, Richardson JR. Coordinated role of voltage-gated sodium channels and the Na+/H+ exchanger in sustaining microglial activation during inflammation. Toxicol Appl Pharmacol, 2013, 273(2): 355-364.
42.	孙阳阳, 骆媛, 王永安. 细胞因子在百草枯中毒致肺纤维化中的作用研究进展. 中国药理学与毒理学杂志, 2021, 35(7): 551-558.
43.	Hou J, Ma T, Cao H, et al. TNF-α-induced NF-κB activation promotes myofibroblast differentiation of LR-MSCs and exacerbates bleomycin-induced pulmonary fibrosis. J Cell Physiol, 2018, 233(3): 2409-2419.
44.	李激文, 钟雨, 刘华, 等. IL-6/STAT3信号通路在百草枯中毒急性肺损伤中作用机制的研究. 右江医学, 2021, 49(3): 172-177.
45.	Hua XF, Li XH, Li MM, et al. Doxycycline attenuates paraquat-induced pulmonary fibrosis by downregulating the TGF-β signaling pathway. J Thorac Dis, 2017, 9(11): 4376-4386.

1. 高利红, 周满红. 百草枯中毒致肺纤维化信号通路研究进展. 中华危重病急救医学, 2021, 33(3): 337-380.
2. Wang J, Zhu Y, Tan J, et al. Lysyl oxidase promotes epithelial-to-mesenchymal transition during paraquat-induced pulmonary fibrosis. Mol Biosyst, 2016, 12(2): 499-507.
3. 王涛, 刘继才, 逄瑜. 羟氯喹的药理学作用机制及安全性风险分析. 中国药物警戒, 2020, 17(07): 385-388.
4. Ramser B, Kokot A, Metze D, et al. Hydroxychloroquine modulates metabolic activity and proliferation and induces autophagic cell death of human dermal fibroblasts. J Invest Dermatol, 2009, 129(10): 2419-2426.
5. Braun S, Ferner M, Kronfeld K, et al. Hydroxychloroquine in children with interstitial (diffuse parenchymal) lung diseases. Pediatr Pulmonol, 2015, 50(4): 410-419.
6. 王超超, 林永强, 蔡小巧, 等. 羟氯喹对IgA肾病患者血清及尿IL-6、TNF-α水平的影响研究. 浙江医学, 2023, 45(5): 514-516, 522.
7. Li D, Yu K, Feng F, et al. Hydroxychloroquine alleviates renal interstitial fibrosis by inhibiting the PI3K/Akt signaling pathway. Biochem Biophys Res Commun, 2022, 610: 154-161.
8. 马爱平, 高云周, 兰文斌, 等. PI3K/Akt/HIF-1α信号通路在博来霉素诱导小鼠肺纤维化中的作用机制研究. 中国呼吸与危重监护杂志, 2016, 15(1): 34-38.
9. Lu Y, Zhong W, Liu Y, et al. Anti-PD-L1 antibody alleviates pulmonary fibrosis by inducing autophagy via inhibition of the PI3K/Akt/mTOR pathway. Int Immunopharmacol, 2022, 104: 108504.
10. 王萍, 李铁刚. 百草枯中毒致小鼠肺纤维化模型的建立. 实用药物与临床, 2020, 23(10): 886-889.
11. 杨伟源, 李理, 郑林鑫, 等. EGFR/Foxo3a/Snail1通路在博来霉素肺纤维化小鼠中的作用机制探讨. 中国呼吸与危重监护杂志, 2020, 19(4): 371-378.
12. Mikawa K, Nishina K, Takao Y, et al. ONO-1714, a nitric oxide synthase inhibitor, attenuates endotoxin-induced acute lung injury in rabbits. Anesth Analg, 2003, 97(6): 1751-1755.
13. 黄海涛, 方静, 曾春晖, 等. 双氢杨梅树皮素对博来霉素诱导小鼠肺纤维化的影响. 中国呼吸与危重监护杂志, 2020, 19(3): 270-275.
14. 刘云华, 周正, 李昌泽, 等. 百草枯中毒致肺纤维化的机制及治疗进展. 毒理学杂志, 2020, 34(2): 166-169,175.
15. 张乙, 陈慧, 崔晋峰. 百草枯诱导肺纤维化机制及治疗研究进展. 临床荟萃, 2019, 34(12): 1134-1137.
16. 李奇, 钱进, 黄琪锋, 等. 百草枯中毒致肺损伤分子机制的研究进展. 海南医学院学报, 2022, 28(4): 309-314, 320.
17. 林琳, 吴开慧, 王文梅. 氯喹/羟氯喹的作用机制及其在口腔疾病中的应用. 口腔疾病防治, 2021, 29(3): 198-201.
18. 崔诚, 么雪婷, 涂思琪, 等. 硫酸羟氯喹的临床药理学研究进展. 中国临床药理学与治疗学, 2020, 25(2): 221-226.
19. Zheng H, Zhang Y, He J, et al. Hydroxychloroquine inhibits macrophage activation and attenuates renal fibrosis after ischemia-reperfusion injury. Front Immunol, 2021, 12: 645100.
20. Ji D, Zhao Q, Qin Y, et al. Germacrone improves liver fibrosis by regulating the PI3K/AKT/mTOR signalling pathway. Cell Biol Int, 2021, 45(9): 1866-1875.
21. Shen H, Wu N, Wang Y, et al. Chloroquine attenuates paraquat-induced lung injury in mice by altering inflammation, oxidative stress and fibrosis. Int Immunopharmacol, 2017, 46: 16-22.
22. Liu L, Ren J, He Z, et al. Cholesterol-modified hydroxychloroquine-loaded nanocarriers in bleomycin-induced pulmonary fibrosis. Sci Rep, 2017, 7(1): 10737.
23. 郑鑫, 纵加强, 王佳新, 等. 基于Wnt/β-catenin信号通路探讨羟氯喹对烟雾吸入致肺纤维化大鼠的作用机制[EB/OL]. 解放军医学院学报, 1-9. (2023-10-23). http://118.89.52.29:8085/kcms/detail/10.1117.R.20231023.0906.002.html.
24. Jiang F, Li S, Jiang Y, et al. Fluorofenidone attenuates paraquat-induced pulmonary fibrosis by regulating the PI3K/Akt/mTOR signaling pathway and autophagy. Mol Med Rep, 2021, 23(6): 405.
25. 周佳宁, 肖倩, 黎芳, 等. 基于PI3K/AKT/mTOR信号通路研究瑶药紫九牛醇提物抗小鼠肝纤维化作用. 中药材, 2022, 45(6): 1470-1474.
26. 潘澎, 刘绍能. PI3K/Akt信号通路与肝纤维化. 临床肝胆病杂志, 2013, 29(5): 389-392, 396.
27. 刘珊珊, 盛春瑞, 杜庆红, 等. 麦门冬汤加减方对特发性肺纤维化小鼠PI3K/AKT/mTOR通路的调控. 现代生物医学进展, 2022, 22(17): 3214-3219, 3224.
28. Hu X, Xu Q, Wan H, et al. PI3K-Akt-mTOR/PFKFB3 pathway mediated lung fibroblast aerobic glycolysis and collagen synthesis in lipopolysaccharide-induced pulmonary fibrosis. Lab Invest, 2020, 100(6): 801-811.
29. Peng J, Xiao X, Li S, et al. Aspirin alleviates pulmonary fibrosis through PI3K/AKT/mTOR-mediated autophagy pathway. Exp Gerontol, 2023, 172: 112085.
30. 林艺凯, 马爱平, 周伟跃, 等. mTOR信号通路在博来霉素诱导小鼠肺纤维化中的作用机制研究. 中国呼吸与危重监护杂志, 2018, 17(2): 178-182.
31. Li X, Ma X, Miao Y, et al. Duvelisib attenuates bleomycin-induced pulmonary fibrosis via inhibiting the PI3K/Akt/mTOR signalling pathway. J Cell Mol Med, 2023, 27(3): 422-434.
32. Qi F, Lv ZD, Huang WD, et al. LncRNA TUG1 promotes pulmonary fibrosis progression via up-regulating CDC27 and activating PI3K/Akt/mTOR pathway. Epigenetics, 2023, 18(1): 2195305.
33. Hill C, Li J, Liu D, et al. Autophagy inhibition-mediated epithelial-mesenchymal transition augments local myofibroblast differentiation in pulmonary fibrosis. Cell Death Dis, 2019, 10(8): 591.
34. He J, Peng H, Wang M, et al. Isoliquiritigenin inhibits TGF-β1-induced fibrogenesis through activating autophagy via PI3K/AKT/mTOR pathway in MRC-5 cells. Acta Biochim Biophys Sin (Shanghai), 2020, 52(8): 810-820.
35. Liu MW, Su MX, Tang DY, et al. 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, 2019, 19(1): 35.
36. Lu J, Qian Y, Jin W, et al. Hypoxia-inducible factor-1α regulates epithelial-to-mesenchymal transition in paraquat-induced pulmonary fibrosis by activating lysyl oxidase. Exp Ther Med, 2018, 15(3): 2287-2294.
37. Zhu Y, Tan J, Xie H, et al. HIF-1α regulates EMT via the Snail and β-catenin pathways in paraquat poisoning-induced early pulmonary fibrosis. J Cell Mol Med, 2016, 20(4): 688-697.
38. Niu Z, Lin J, Hao C, et al. Glycyrrhizic acid attenuates pulmonary fibrosis of silicosis by inhibiting the interaction between HMGB1 and BRG1 through PI3K/Akt/mTOR pathway. Int J Environ Res Public Health, 2022, 19(14): 8743.
39. Wang Y, Men M, Xie B, et al. Inhibition of PKR protects against H2O2-induced injury on neonatal cardiac myocytes by attenuating apoptosis and inflammation. Sci Rep, 2016, 6: 38753.
40. 何文涓, 袁志坚, 黄寅, 等. 芹菜素对百草枯致小鼠肺纤维化保护作用及机制研究. 药物评价研究, 2021, 44(8): 1660-1665.
41. Hossain MM, Sonsalla PK, Richardson JR. Coordinated role of voltage-gated sodium channels and the Na+/H+ exchanger in sustaining microglial activation during inflammation. Toxicol Appl Pharmacol, 2013, 273(2): 355-364.
42. 孙阳阳, 骆媛, 王永安. 细胞因子在百草枯中毒致肺纤维化中的作用研究进展. 中国药理学与毒理学杂志, 2021, 35(7): 551-558.
43. Hou J, Ma T, Cao H, et al. TNF-α-induced NF-κB activation promotes myofibroblast differentiation of LR-MSCs and exacerbates bleomycin-induced pulmonary fibrosis. J Cell Physiol, 2018, 233(3): 2409-2419.
44. 李激文, 钟雨, 刘华, 等. IL-6/STAT3信号通路在百草枯中毒急性肺损伤中作用机制的研究. 右江医学, 2021, 49(3): 172-177.
45. Hua XF, Li XH, Li MM, et al. Doxycycline attenuates paraquat-induced pulmonary fibrosis by downregulating the TGF-β signaling pathway. J Thorac Dis, 2017, 9(11): 4376-4386.

Previous Article
Annexin A1 derived from umbilical cord mesenchymal stem cells protects against lipopolysaccharide-induced acute lung injury
Next Article
无并发症超长期留置经口气管插管一例

Chinese Journal of Respiratory and Critical Care Medicine

Effect of hydroxychloroquine sulphate on paraquat-induced lung fibrosis in mice via PI3K/AKt/mTOR signalling pathway

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content