Vanillic acid alleviates lipopolysaccharides-induced endoplasmic reticulum stress and inflammation in human lung fibroblasts by `inhibiting MAPK and NF-κB pathways

Main Article Content

Jihua Zhao
Yao Yang


vanillic acid, lipopolysaccharides, inflammation, endoplasmic reticulum stress, human lung fibroblasts, MAPK, NF-κB, pneumonia


Persistent endoplasmic reticulum stress promotes aberrant inflammation and induces cell death, and inflammation is implicated in the pathogenesis of pneumonia. Vanillic acid exerts pharmacological activities, such as anti-inflammatory, antimicrobial, and antioxidant effects. However, the role of vanillic acid in pneumonia has not been elucidated yet. Human lung fibroblasts (WI-38 and MRC-5) were incubated with different concentrations of lipopolysaccharides to mimic the cell model of pneumonia. Lipopolysaccharides-treated lung fibroblasts were then incubated with different concentrations of vanillic acid. Cell viability and apoptosis were detected by MTT assay and flow cytometry, respectively. Quantitative real-time polymerase chain reaction and enzyme-linked-immunosorbent serologic assay were used to measure the levels of inflammatory factors. Western blot assay was used to detect endoplasmic reticulum stress and downstream pathway. Lipopolysaccharides induced decrease of cell viability in WI-38 and MRC-5 whereas vanillic acid increased cell viability of lipopolysaccharides-treated lung fibroblasts. Lipopolysaccharides-induced increase of cell apoptosis in lung fibroblasts was suppressed by vanillic acid through up-regulation of BCL2, and down-regulation of BCL2 associated X (BAX) and cleaved caspase-3. Vanillic acid reduced levels of tumor necrosis factor-α (TNF-α), Interleukin 6 (IL-6), and IL-1β in lipopolysaccharides-treated lung fibroblasts. Protein expression of glucose-regulated protein 78 (GRP78), X-box binding protein 1 (XBP-1), activating transcription factor-6 (ATF-6), ATF-4, and C/EBP homologous protein (CHOP) in lung fibroblasts were up-regulated by lipopolysaccharides while reduced by vanillic acid. Vanillic acid attenuated lipopolysaccharides-induced decrease of IκBα and increase of p-IκBα, p-p65, p-ERK, and p-JNK in fibroblasts. Vanillic acid exerted anti-inflammatory effect against lipopolysaccharides-induced human lung fibroblasts by inhibiting mitogen-activated protein kinase and nuclear factor kappa B pathways.

Abstract 528 | PDF Downloads 446 HTML Downloads 361 XML Downloads 52


Amin, F.U., Shah, S.A. and Kim, M.O., 2017. Vanillic acid attenuates Aβ1-42-induced oxidative stress and cognitive impairment in mice. Scientific Reports 7: 40753. 10.1038/srep40753

Bai, F., Fang, L., Hu, H., Yang, Y., Feng, X. and Sun, D., 2019. Vanillic acid mitigates the ovalbumin (OVA)-induced asthma in rat model through prevention of airway inflammation. Bioscience, Biotechnology, and Biochemistry 83: 531–537. 10.1080/09168451.2018.1543015

Calixto-Campos, C.S., 2015. Vanillic acid inhibits inflammatory pain by inhibiting neutrophil recruitment, oxidative stress, cytokine production, and NFκB activation in mice. Journal of Natural Products 78: 1799–1808. 10.1021/acs.jnatprod.5b00246

Chi, G., Wei, M., Xie, X., Soromou, L.W., Liu, F. and Zhao, S., 2012. Suppression of MAPK and NF-κB pathways by limonene contributes to attenuation of lipopolysaccharide-induced inflammatory responses in acute lung injury. Inflammation 36(2): 501–511. 10.1007/s10753-012-9571-1

Dassner, M.A., Nicolau, P.D. and Girotto, E.J., 2017. Management of pneumonia in the pediatric critical care unit: an area for antimicrobial stewardship. Current Pediatric Reviews 13: 49–66. 10.2174/1573396312666161205102221

Huang, C.-Y., Deng, J.-S., Huang, W.-C., Jiang, W.-P. and Huang, G.-J., 2020. Attenuation of lipopolysaccharide-induced acute lung injury by hispolon in mice, through regulating the TLR4/PI3K/Akt/mTOR and Keap1/Nrf2/HO-1 pathways, and suppressing oxidative stress-mediated ER stress-induced apoptosis and autophagy. Nutrients 12: 1742.

Huang, X., Xi, Y., Mao, Z., Chu, X., Zhang, R., Ma, X., Ni, B., Cheng, H. and You, H., 2019. Vanillic acid attenuates cartilage degeneration by regulating the MAPK and PI3K/AKT/NF-κB pathways. European Journal of Pharmacology 859: 172481. 10.1016/j.ejphar.2019.172481

Hur, I., Ozkan, S., Halici, A., Abatay, K., Usul, E., Cetin, E. and Aydin, F.N., 2020. Role of plasma presepsin, procalcitonin and C-reactive protein levels in determining the severity and mortality of community-acquired pneumonia in the emergency department. Signa Vitae 16: 61–68.

Ji, G., Sun, R., Hu, H., Xu, F., Yu, X., Veeraraghavan, V., Krishna Mohan, S. and Chi, X., 2020. Vannilic acid ameliorates hyperglycemia-induced oxidative stress and inflammation in streptozotocin-induced diabetic rats. Journal of King Saud University–Science 32(7): 2905–2911. 10.1016/j.jksus.2020.04.010

Kim, H.J., Jeong, J.S., Kim, S.R., Park, S.Y., Chae, H.J. and Lee, Y.C., 2013. Inhibition of endoplasmic reticulum stress alleviates lipopolysaccharide-induced lung inflammation through modulation of NF-κB/HIF-1α signaling pathway. Scientific Reports 3: 1142–1142. 10.1038/srep01142

Kim, M.-C., Kim, S.-J., Kim, D.-S., Jeon, Y.-D., Park, S.J., Lee, H.S., Um, J.-Y. and Hong, S.-H., 2011. Vanillic acid inhibits inflammatory mediators by suppressing NF-κB in lipopolysaccharide-stimulated mouse peritoneal macrophages. Immunopharmacology and Immunotoxicology 33: 525–532. 10.3109/08923973.2010.547500

Köseler, A., Sabirli, R., Gören, T., Türkçüer, I. and Kurt, Ö., 2020. Endoplasmic reticulum stress markers in SARS-COV-2 infection and pneumonia: case-control study. In Vivo (Athens, Greece) 34: 1645–1650. 10.21873/invivo.11956

Marangu, D. and Zar, H.J., 2019. Childhood pneumonia in low-and-middle-income countries: an update. Paediatric Respiratory Reviews 32: 3–9. 10.1016/j.prrv.2019.06.001

Mizgerd, J.P., 2018. Inflammation and pneumonia: why are some more susceptible than others? Clinics in Chest Medicine 39: 669–676. 10.1016/j.ccm.2018.07.002

Müller-Redetzky, H., Lienau, J., Suttorp, N. and Witzenrath, M., 2015. Therapeutic strategies in pneumonia: going beyond antibiotics. European Respiratory Review 24: 516. 10.1183/16000617.0034-2015

Nguyen, T., Binh, T., Kusunoki, R., Pham, T., Nguyen, H.Y., Tuân, N., Kanaori, K. and Kamei, K., 2020. Effects of Launaea sarmentosa extract on lipopolysaccharide-induced inflammation via suppression of NF-κB/MAPK signaling and Nrf2 activation. Nutrients 12(9): 2586. 10.3390/nu12092586

Quan, B., Zhang, H. and Xue, R., 2019. miR-141 alleviates LPS-induced inflammation injury in WI-38 fibroblasts by up-regulation of NOX2. Life Sciences 216: 271–278. 10.1016/j.lfs.2018.11.056

Scoditti, E., Massaro, M., Garbarino, S. and Toraldo, D.M., 2019. Role of diet in chronic obstructive pulmonary disease prevention and treatment. Nutrients 11: 1357. 10.3390/nu11061357

Seki, H., Fukunaga, K., Arita, M., Arai, H., Nakanishi, H., Taguchi, R., Miyasho, T., Takamiya, R., Asano, K., Ishizaka, A., Takeda, J. and Levy, B.D., 2010. The anti-inflammatory and proresolving mediator resolvin E1 protects mice from bacterial pneumonia and acute lung injury. Journal of Immunology (Baltimore, MD) 184: 836–843. 10.4049/jimmunol.0901809

Siddiqui, S., Kamal, A., Khan, F., Jamali, K.S. and Saify, Z.S., 2019. Gallic and vanillic acid suppress inflammation and promote myelination in an in vitro mouse model of neurodegeneration. Molecular Biology Reports 46: 997–1011. 10.1007/s11033-018-4557-1

Simopoulos, A.P., 2021. Genetic variation, diet, inflammation, and the risk for COVID-19. Lifestyle Genomics 14: 37–42. 10.1159/000513886

Tang, X., Wang, T., Qiu, C., Zheng, F., Xu, J. and Zhong, B., 2020. Long non-coding RNA (lncRNA) CRNDE regulated lipopolysaccharides (LPS)-induced MRC-5 inflammation injury through targeting MiR-141. Medical Science Monitor: International Medical Journal of Experimental and Clinical Research 26: e920928–e920928. 10.12659/MSM.920928

Ullah, R., Ikram, M., Park, T.J., Ahmad, R., Saeed, K., Alam, S.I., Rehman, I.U., Khan, A., Khan, I., Jo, M.G. and Kim, M.O., 2020. Vanillic acid, a bioactive phenolic compound, counteracts LPS-induced neurotoxicity by regulating c-Jun N-terminal kinase in mouse brain. International Journal of Molecular Sciences 22: 361. 10.3390/ijms22010361

Xiao, K., Liu, C., Tu, Z., Xu, Q., Chen, S., Zhang, Y., Wang, X., Zhang, J., Hu, C.-A.A. and Liu, Y., 2020. Activation of the NF-κB and MAPK signaling pathways contributes to the inflammatory responses, but not cell injury in IPEC-1 cells challenged with hydrogen peroxide. Oxidative Medicine and Cellular Longevity 2020: 5803639. 10.1155/2020/5803639

Zeng, M., Sang, W., Chen, S., Chen, R., Zhang, H., Xue, F., Li, Z., Liu, Y., Gong, Y., Zhang, H. and Kong, X., 2017. 4-PBA inhibits LPS-induced inflammation through regulating ER stress and autophagy in acute lung injury models. Toxicology Letters 271: 26–37. 10.1016/j.toxlet.2017.02.023

Zhang, K. and Kaufman, R.J., 2008. From endoplasmic-reticulum stress to the inflammatory response. Nature 454: 455–462. 10.1038/nature07203