Korean J Physiol Pharmacol.  2024 Nov;28(6):549-558. 10.4196/kjpp.2024.28.6.549.

Salidroside attenuates sepsis-induced acute lung injury by inhibiting ferroptosis-dependent pathway

• Zhen L
• Hou M
• Wang S

• KMID: 2560734
• DOI: http://doi.org/10.4196/kjpp.2024.28.6.549

Abstract

Sepsis triggers a systemic inflammatory response that can lead to acute lung injury (ALI). Salidroside (SAL) has many pharmacological activities such as antiinflammatory and anti-oxidation. The objective of the study was to explore the mechanism of SAL on ALI caused by sepsis. A model of ALI in septic mice was established by cecal ligation and puncture. Following SAL treatment, the effect of SAL on the ferroptosis pathway in mice was analyzed. The pathological damage of lung tissue, the levels of inflammatory factors and apoptosis in bronchoalveolar lavage fluid (BALF) of mice were evaluated, and the changes of gene expression level and metabolite content abundance were explored by combining transcriptomics and metabolomics analysis. The effect of SAL on ferroptosis in mice with lung injury was observed by intraperitoneal injection of ferroptosis activator Erastin or ferroptosis inhibitor Ferrostatin-1 to promote or inhibit ferroptosis in mice. SAL significantly alleviated the pathological damage of lung tissue, decreased the number of TUNEL positive cells and the levels of TNF-α, IL-1β, IL-6 in BALF, and increased the level of IL-10 in lung injury mice. Moreover, the Fe 2+ content and malondialdehyde decreased significantly, the reactive oxygen species and glutathione content increased significantly, and the arachidonic acid metabolites 20-hydroxyeicosatetraenoic acid (20-HETE), (5Z, 8Z, 10E, 14Z)-12-Oxoeicosa-5,8,10,14-tetraenoic acid (12-OxOETE), (5Z, 8Z, 10E, 14Z)-(12S)-12-Hydroxyeicosa-5,8,10,14-tetraenoic acid (12(S)-HETE), (5Z, 8Z, 14Z)-11,12-Dihydroxyeicosa-5,8,14-trienoic acid (11,12-DHET), (5Z, 11Z, 14Z)-8,9-Dihydroxyeicosa-5,11,14-trienoic acid, Leukotriene B4, Leukotriene D4 were significantly up-regulated after SAL treatment. Salidroside alleviates ALI caused by sepsis by inhibiting ferroptosis.

Keyword

Acute lung injury; Arachidonic acid; Ferroptosis; Lipid metabolism; Rhodioloside; Sepsis

Figure

• 

  Fig. 1 SAL ameliorates sepsis-induced acute lung injury. (A) H&E staining was used to analyze the morphological changes of lung tissue in mice (x20 and x200). Yellow arrows point to alveoli, red arrows to interstitial tissue, and black arrows to inflammatory infiltrates. (B) Pulmonary edema was assessed by analyzing the lung W/D ratio. (C) TUNEL staining was used to evaluate apoptosis (x200). (D) The levels of inflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-10) in BALF of mice were measured by ELISA. N = 6. Values are presented as mean ± SD. SAL, salidroside; W/D ratio, wet weight/dry weight ratio; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; TNF-α, tumor necrosis factor-α; IL, interleukin; BALF, bronchoalveolar lavage fluid; CLP, cecal ligation and puncture. *p < 0.05, **p < 0.01, ***p < 0.001.

• 

  Fig. 2 Effects of SAL on ferroptosis pathway and metabolites in septic mice. Transcriptome and metabolomics analysis of lung tissue in Sham group, CLP group and salidroside (100 mg/kg) treatment group. (A) Log2(FC) was used as the ordinate, and the grouping name was used as the abscissa to display the differentially expressed genes between groups. Green represented down-regulated genes, red represented up-regulated genes. (B) GSEA analysis was performed on all genes in the Sham group and the CLP group and the CLP and CLP + SAL 100 groups using the KEGG database. The positive and negative ES values indicated the correlation between the pathway and the comparison group. The positive ES value indicated that the pathway was positively correlated with the comparison group compared with the control group, and the negative ES value indicated that the pathway was negatively correlated with the comparison group compared with the control group. (C) OPLS-DA was used to analyze the loading map to find out the metabolites that contributed the most to the change of metabolite patterns between the Sham group and the CLP group, the CLP group and the CLP + SAL 100 group. The farther away from the origin in the abscissa direction, the greater the contribution of the variables to the distinction between the two groups of samples. SAL, salidroside; CLP, cecal ligation and puncture; GSEA, gene set enrichment analysis; KEGG, Kyoto Encyclopedia of Genes and Genomes; ES, enrichment score; OPLS-DA, orthogonal partial least squares discriminant analysis.

• 

  Fig. 3 Relationship between ferroptosis-related DEGs and metabolites in mice with sepsis-induced acute lung injury. (A) The expression levels of DEGs related to ferroptosis and arachidonic acid metabolism were displayed by heat map. (B) The correlation between arachidonic acid metabolism metabolites 11,12-dihydroxy-5Z,8Z,14Z-eicosatrienoic acid (11,12-DHET), 20-HETE and Alox15. (C) Arachidonic acid metabolism related DEGs and DMs schematic diagram. The rectangle is divided into two color blocks, left: Sham vs. CLP, right: CLP vs. CLP + SAL 100. 20-HETE, 20-hydroxyeicosatetraenoic acid; 12-OxoETE, (5Z, 8Z, 10E, 14Z)-12-Oxoeicosa-5,8,10,14-tetraenoic acid; 12(S)-HETE, (5Z, 8Z, 10E, 14Z)-(12S)-12-Hydroxyeicosa-5,8,10,14-tetraenoic acid; 11,12-DHET, (5Z, 8Z, 14Z)-11,12-Dihydroxyeicosa-5,8,14-trienoic acid; 8,9-DHET, (5Z, 11Z, 14Z)-8,9-Dihydroxyeicosa-5,11,14-trienoic acid; DEGs, differentially expressed genes; SAL, salidroside; CLP, cecal ligation and puncture; KEGG, Kyoto Encyclopedia of Genes and Genomes; DMs, differential metabolites.

• 

  Fig. 4 SAL alleviates lung injury in septic mice by inhibiting ferroptosis. (A) H&E staining was used to analyze the morphological changes of lung tissue in mice (x20 and x200). Yellow arrows point to alveoli, red arrows to interstitial tissue, and black arrows to inflammatory infiltrates. (B) Pulmonary edema was evaluated by analyzing the lung W/D ratio. (C) TUNEL staining to evaluate apoptosis (x200). (D) The levels of TNF-α, IL-1β, IL-6 and IL-10 in BALF were measured by ELISA. (E) ROS level was detected by immunofluorescence (x200). (F) The content of Fe2+, glutathione (GSH) and malondialdehyde (MDA) were detected by kit. N = 6. Values are presented as mean ± SD. SAL, salidroside; W/D ratio, wet weight/dry weight ratio; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; TNF-α, tumor necrosis factor-α; IL, interleukin; BALF, bronchoalveolar lavage fluid; ROS, reactive oxygen species; CLP, cecal ligation and puncture; Fer-1, ferrostatin-1. *p < 0.05, **p < 0.01, ***p < 0.001.

Reference

1. Jiang W, Ma C, Bai J, Du X. 2022; Macrophage SAMSN1 protects against sepsis-induced acute lung injury in mice. Redox Biol. 56:102432. Erratum in: Redox Biol. 2024;70:103036. DOI: 10.1016/j.redox.2024.103036. PMID: 35981417. PMCID: PMC9418554.

2. Hwang JS, Kim KH, Park J, Kim SM, Cho H, Lee Y, Han IO. 2019; Glucosamine improves survival in a mouse model of sepsis and attenuates sepsis-induced lung injury and inflammation. J Biol Chem. 294:608–622. DOI: 10.1074/jbc.RA118.004638. PMID: 30455348. PMCID: PMC6333887.

3. Zhang J, Zheng Y, Wang Y, Wang J, Sang A, Song X, Li X. 2022; YAP1 alleviates sepsis-induced acute lung injury via inhibiting ferritinophagy-mediated ferroptosis. Front Immunol. 13:884362. DOI: 10.3389/fimmu.2022.884362. PMID: 35979359. PMCID: PMC9376389.

4. Ingold I, Berndt C, Schmitt S, Doll S, Poschmann G, Buday K, Roveri A, Peng X, Porto Freitas F, Seibt T, Mehr L, Aichler M, Walch A, Lamp D, Jastroch M, Miyamoto S, Wurst W, Ursini F, Arnér ESJ, Fradejas-Villar N, et al. 2018; Selenium utilization by GPX4 is required to prevent hydroperoxide-induced ferroptosis. Cell. 172:409–422.e21. DOI: 10.1016/j.cell.2017.11.048. PMID: 29290465.

5. Alim I, Caulfield JT, Chen Y, Swarup V, Geschwind DH, Ivanova E, Seravalli J, Ai Y, Sansing LH, Ste Marie EJ, Hondal RJ, Mukherjee S, Cave JW, Sagdullaev BT, Karuppagounder SS, Ratan RR. 2019; Selenium drives a transcriptional adaptive program to block ferroptosis and treat stroke. Cell. 177:1262–1279.e25. DOI: 10.1016/j.cell.2019.03.032. PMID: 31056284.

6. Kim MJ, Yun GJ, Kim SE. 2021; Metabolic regulation of ferroptosis in cancer. Biology (Basel). 10:83. DOI: 10.3390/biology10020083. PMID: 33499222. PMCID: PMC7911352.

7. Molchanova AY, Rjabceva SN, Melik-Kasumov TB, Pestov NB, Angelova PR, Shmanai VV, Sharko OL, Bekish AV, James G, Park HG, Udalova IA, Brenna JT, Shchepinov MS. 2022; Deuterated arachidonic acid ameliorates lipopolysaccharide-induced lung damage in mice. Antioxidants (Basel). 11:681. DOI: 10.3390/antiox11040681. PMID: 35453366. PMCID: PMC9027010.

8. Xu F, Xu J, Xiong X, Deng Y. 2019; Salidroside inhibits MAPK, NF-κB, and STAT3 pathways in psoriasis-associated oxidative stress via SIRT1 activation. Redox Rep. 24:70–74. DOI: 10.1080/13510002.2019.1658377. PMID: 31495284. PMCID: PMC6748574.

9. Zhong Z, Han J, Zhang J, Xiao Q, Hu J, Chen L. 2018; Pharmacological activities, mechanisms of action, and safety of salidroside in the central nervous system. Drug Des Devel Ther. 12:1479–1489. DOI: 10.2147/DDDT.S160776. PMID: 29872270. PMCID: PMC5973445.

10. Huang X, Xue H, Ma J, Zhang Y, Zhang J, Liu Y, Qin X, Sun C. 2019; Salidroside ameliorates Adriamycin nephropathy in mice by inhibiting β-catenin activity. J Cell Mol Med. 23:4443–4453. DOI: 10.1111/jcmm.14340. PMID: 30993911. PMCID: PMC6533469.

11. Hasin Y, Seldin M, Lusis A. 2017; Multi-omics approaches to disease. Genome Biol. 18:83. DOI: 10.1186/s13059-017-1215-1. PMID: 28476144. PMCID: PMC5418815.

12. Xie X, Liao J, Ai Y, Gao J, Zhao J, Qu F, Xu C, Zhang Z, Wen W, Cui H, Wang H. 2021; Pi-Dan-Jian-Qing decoction ameliorates type 2 diabetes mellitus through regulating the gut microbiota and serum metabolism. Front Cell Infect Microbiol. 11:748872. DOI: 10.3389/fcimb.2021.748872. PMID: 34938667. PMCID: PMC8685325.

13. Bylesjö M, Rantalainen M, Cloarec O, Nicholson JK, Holmes E, Trygg J. 2006; OPLS discriminant analysis: combining the strengths of PLS-DA and SIMCA classification. J Chemom. 20:341–351. DOI: 10.1002/cem.1006.

14. Tang H, Gao L, Mao J, He H, Liu J, Cai X, Lin H, Wu T. 2016; Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-κB and TGF-β1/Smad-2/-3 pathways. Cell Stress Chaperones. 21:239–249. DOI: 10.1007/s12192-015-0654-4. PMID: 26577463. PMCID: PMC4786523.

15. Liu MW, Su MX, Qin LF, Liu X, Tian ML, Zhang W, Wang YH. 2014; Effect of salidroside on lung injury by upregulating peroxisome proliferator-activated receptor γ expression in septic rats. Exp Ther Med. 7:1446–1456. DOI: 10.3892/etm.2014.1629. PMID: 24926325. PMCID: PMC4043580.

16. Wang Y, Xu CF, Liu YJ, Mao YF, Lv Z, Li SY, Zhu XY, Jiang L. 2017; Salidroside attenuates ventilation induced lung injury via SIRT1-dependent inhibition of NLRP3 inflammasome. Cell Physiol Biochem. 42:34–43. DOI: 10.1159/000477112. PMID: 28490015.

17. Fan H, Su BJ, Le JW, Zhu JH. 2022; Salidroside protects acute kidney injury in septic rats by inhibiting inflammation and apoptosis. Drug Des Devel Ther. 16:899–907. DOI: 10.2147/DDDT.S361972. PMID: 35386851. PMCID: PMC8978577.

18. Jiang L, Xu L, Zheng L, Wang Y, Zhuang M, Yang D. 2022; Salidroside attenuates sepsis-associated acute lung injury through PPP1R15A mediated endoplasmic reticulum stress inhibition. Bioorg Med Chem. 71:116865. DOI: 10.1016/j.bmc.2022.116865. PMID: 35985062.

19. Wang Y, Chen Z, Luo J, Zhang J, Sang AM, Cheng ZS, Li XY. 2023; Salidroside postconditioning attenuates ferroptosis-mediated lung ischemia-reperfusion injury by activating the Nrf2/SLC7A11 signaling axis. Int Immunopharmacol. 115:109731. Erratum in: Int Immunopharmacol. DOI: 10.1016/j.intimp.2023.110002. PMID: 36907990.

20. Chen X, Li J, Kang R, Klionsky DJ, Tang D. 2021; Ferroptosis: machinery and regulation. Autophagy. 17:2054–2081. DOI: 10.1080/15548627.2020.1810918. PMID: 32804006. PMCID: PMC8496712.

21. Kagan VE, Mao G, Qu F, Angeli JP, Doll S, Croix CS, Dar HH, Liu B, Tyurin VA, Ritov VB, Kapralov AA, Amoscato AA, Jiang J, Anthonymuthu T, Mohammadyani D, Yang Q, Proneth B, Klein-Seetharaman J, Watkins S, Bahar I, et al. 2017; Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis. Nat Chem Biol. 13:81–90. DOI: 10.1038/nchembio.2238. PMID: 27842066. PMCID: PMC5506843.

22. Tang C, Tang Y, Wang Q, Chu D, Zhou J, Zhou Y. 2022; Yangyinqingfei decoction attenuates PM_2.5-induced lung injury by enhancing arachidonic acid metabolism. Front Pharmacol. 13:1056078. DOI: 10.3389/fphar.2022.1056078. PMID: 36467030. PMCID: PMC9708729.

23. Lewis RA, Austen KF, Soberman RJ. 1990; Leukotrienes and other products of the 5-lipoxygenase pathway. Biochemistry and relation to pathobiology in human diseases. N Engl J Med. 323:645–655. DOI: 10.1056/NEJM199009063231006. PMID: 2166915.

24. Lee JY, Nam M, Son HY, Hyun K, Jang SY, Kim JW, Kim MW, Jung Y, Jang E, Yoon SJ, Kim J, Kim J, Seo J, Min JK, Oh KJ, Han BS, Kim WK, Bae KH, Song J, Kim J, et al. 2020; Polyunsaturated fatty acid biosynthesis pathway determines ferroptosis sensitivity in gastric cancer. Proc Natl Acad Sci U S A. 117:32433–32442. DOI: 10.1073/pnas.2006828117. PMID: 33288688. PMCID: PMC7768719.

25. Sun Y, Chen P, Zhai B, Zhang M, Xiang Y, Fang J, Xu S, Gao Y, Chen X, Sui X, Li G. 2020; The emerging role of ferroptosis in inflammation. Biomed Pharmacother. 127:110108. DOI: 10.1016/j.biopha.2020.110108. PMID: 32234642.

26. Gao M, Deng J, Liu F, Fan A, Wang Y, Wu H, Ding D, Kong D, Wang Z, Peer D, Zhao Y. 2019; Triggered ferroptotic polymer micelles for reversing multidrug resistance to chemotherapy. Biomaterials. 223:119486. DOI: 10.1016/j.biomaterials.2019.119486. PMID: 31520887.

27. Reichert CO, de Freitas FA, Sampaio-Silva J, Rokita-Rosa L, Barros PL, Levy D, Bydlowski SP. 2020; Ferroptosis mechanisms involved in neurodegenerative diseases. Int J Mol Sci. 21:8765. DOI: 10.3390/ijms21228765. PMID: 33233496. PMCID: PMC7699575.

28. Zhao J, Piao X, Wu Y, Liang S, Han F, Liang Q, Shao S, Zhao D. 2020; Cepharanthine attenuates cerebral ischemia/reperfusion injury by reducing NLRP3 inflammasome-induced inflammation and oxidative stress via inhibiting 12/15-LOX signaling. Biomed Pharmacother. 127:110151. DOI: 10.1016/j.biopha.2020.110151. PMID: 32559840.

29. Łuczaj W, Gęgotek A, Skrzydlewska E. 2017; Antioxidants and HNE in redox homeostasis. Free Radic Biol Med. 111:87–101. DOI: 10.1016/j.freeradbiomed.2016.11.033. PMID: 27888001.