Phillygenin attenuates LPS-induced acute lung injury of newborn mice in infantile pneumonia

Purpose: The aim of this study was to assess the effect of phillygenin (PHI) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and understand its underlying mechanisms.

Methods: Mice were separated into four different groups at random, including LPS, LPS+ PHI (5 mg/kg), LPS + PHI (50 mg/kg) and control group. The two LPS + PHI groups were intraperitoneally administered with PHI after LPS intratracheal administered for 1 h. Subsequently, the lung tissues of different groups were collected and evaluated by H&E staining and W/D (W/D) ratio. The inflammatory cytokines in BALF or lung tissue were also assessed. Western blot assay was applied to examine the expressions of TLR4, MyD88, and NF-κB.

Results: The ameliorated pathological changes and lung W/D ratio demonstrated that PHI dramatically suppressed the lung injury levels. PHI strikingly reduced the number of inflammatory cell counts and total protein concentration in BALF. In addition, PHI attenuated expression of IL-1β and TNF-α in BALF and lung tissue. Furthermore, it was confirmed that PHI alleviated LPS-induced ALI via TLR4/MyD88/NF-κB pathway.

Conclusions: Together the above results show that PHI attenuates LPS-induced ALI via inactivation of TLR4/MyD88/NF-κB pathway in newborn mice.

Phillygenin; Acute lung injury; TLR4; MyD88; NF-κB

[1] de Luca D, Piastra M, Tosi F, Pulitano S, Mancino A, Genovese O, et al. Pharmacological therapies for pediatric and neonatal ALI/ARDS: an evidence-based review. Current Drug Targets. 2012; 13: 906–916.

[2] Bellani G, Laffey JG, Pham T, Fan E, Brochard L, Esteban A, et al. Epidemiology, patterns of care, and mortality for patients with acute respiratory distress syndrome in intensive care units in 50 countries. The Journal of the American Medical Association. 2016; 315: 788.

[3] Butt Y, Kurdowska A, Allen TC. Acute lung injury: a clinical and molecular review. Archives of Pathology & Laboratory Medicine. 2016; 140: 345–350.

[4] Mowery NT, Terzian WTH, Nelson AC. Acute lung injury. Current Problems in Surgery. 2020; 57: 100777.

[5] Mendez JL, Hubmayr RD. New insights into the pathology of acute respiratory failure. Current Opinion in Critical Care. 2005; 11: 29–36.

[6] Avlas O, Fallach R, Shainberg A, Porat E, Hochhauser E. Toll-like receptor 4 stimulation initiates an inflammatory response that decreases cardiomyocyte contractility. Antioxidants & Redox Signaling. 2011; 15: 1895–1909.

[7] Liu L, Gu H, Liu H, Jiao Y, Li K, Zhao Y, et al. Protective effect of resveratrol against IL-1β-induced inflammatory response on human osteoarthritic chondrocytes partly via the TLR4/MyD88/NF-κB signaling pathway: an “in vitro study”. International Journal of Molecular Sciences. 2014; 15: 6925–6940.

[8] Hou Y, Luo S, Zhang Y, Jia Y, Li H, Xiao C, et al. Contrasting effects of acute and long-term corticosterone treatment on amyloid-β, beta-secretase 1 expression, and nuclear factor kappa B nuclear translocation. Journal of Integrative Neuroscience. 2019; 18: 393–400.

[9] Castoldi A, Braga TT, Correa-Costa M, Aguiar CF, Bassi ÊJ, Correa-Silva R, et al. TLR2, TLR4 and the MyD88 signaling pathway are crucial for neutrophil migration in acute kidney injury induced by sepsis. PLoS ONE. 2012; 7: e37584.

[10] Hung CH, Tsai Y, Li K. Phenolic antioxidants isolated from the flowers of Osmanthus fragrans. Molecules. 2012; 17: 10724–10737.

[11] Lee D, Lee S, Bang M, Park H, Lee T, Kim Y, et al. Lignans from the flowers of Osmanthus fragrans var. aurantiacus and their inhibition effect on no production. Archives of Pharmacal Research. 2011; 34: 2029–2035.

[12] Wang H, Cao Z-R. Anti-inflammatory effects of (-)-epicatechin in lipopolysaccharide-stimulated raw 264.7 macrophages. Tropical Journal of Pharmaceutical Research. 2014; 13: 1415.

[13] Hu N, Wang C, Dai X, Zhou M, Gong L, Yu L, et al. Phillygenin inhibits LPS-induced activation and inflammation of LX2 cells by TLR4/MyD88/NF-κB signaling pathway. Journal of Ethnopharmacol-ogy. 2020; 248: 112361.

[14] National Research Council Committee for the Update of the Guide for the C, Use of Laboratory A. The national academies collection: reports funded by National Institutes of Health. Guide for the care and use of laboratory animals. Washington: National Academies Press. 2011.

[15] Ju M, Liu B, He H, Gu Z, Liu Y, Su Y, et al. MicroRNA-27a alleviates LPS-induced acute lung injury in mice via inhibiting inflammation and apoptosis through modulating TLR4/MyD88/NF-κB pathway. Cell Cycle. 2018; 17: 2001–2018.

[16] Wang N, Geng C, Sun H, Wang X, Li F, Liu X. Hesperetin ameliorates lipopolysaccharide-induced acute lung injury in mice through regulating the TLR4-MyD88-NF-κB signaling pathway. Archives of Pharmacal Research. 2019; 42: 1063–1070.

[17] Jiang Q, Yi M, Guo Q, Wang C, Wang H, Meng S, et al. Protective effects of polydatin on lipopolysaccharide-induced acute lung injury through TLR4-MyD88-NF-κB pathway. International Immunopharmacology. 2015; 29: 370–376.

[18] Tianzhu Z, Shumin W. Esculin inhibits the inflammation of LPS-induced acute lung injury in mice via regulation of TLR/NF-κB pathways. Inflammation. 2015; 38: 1529–1536.

[19] Tao W, Su Q, Wang H, Guo S, Chen Y, Duan J, et al. Platycodin D attenuates acute lung injury by suppressing apoptosis and inflammation in vivo and in vitro. International Immunopharmacology. 2016; 27: 138–147.

[20] Cheng N, Ren N, Gao H, Lei X, Zheng J, Cao W. Antioxidant and hepatoprotective effects of Schisandra chinensis pollen extract on CCl4-induced acute liver damage in mice. Food and Chemical Toxicology. 2013; 55: 234–240.

[21] Kim SR, Lee MK, Koo KA, Kim SH, Sung SH, Lee NG, et al. Dibenzocyclooctadiene lignans from Schisandra chinensis protect primary cultures of rat cortical cells from glutamate-induced toxicity. Journal of Neuroscience Research. 2004; 76: 397–405.

[22] Sowndhararajan K, Deepa P, Kim M, Park SJ, Kim S. An overview of neuroprotective and cognitive enhancement properties of lignans from Schisandra chinensis. Biomedicine & Pharmacotherapy. 2018; 97: 958–968.

[23] Brandenberger C, Kling KM, Vital M, Christian M. The role of pulmonary and systemic immunosenescence in acute lung injury. Aging and Disease. 2018; 9: 553–565.

[24] Zhi H, Zhu H, Zhang Y, Lu Y, Li H, Chen D. In vivo effect of quantified flavonoids-enriched extract of Scutellaria baicalensis root on acute lung injury induced by influenza a virus. Phytomedicine. 2019; 57: 105–116.

[25] Luo Y, Pang X, Ansari AR, Wu X, Li H, Zhang Z, et al. Visfatin exerts immunotherapeutic effects in lipopolysaccharide-induced acute lung injury in murine model. Inflammation. 2020; 43: 109–122.

[26] Zhao G, Zhang T, Ma X, Jiang K, Wu H, Qiu C, et al. Oridonin attenuates the release of pro-inflammatory cytokines in lipopolysaccharide-induced RAW264.7 cells and acute lung injury. Oncotarget. 2017; 8: 68153–68164.

[27] Chen C, Shi L, Li Y, Wang X, Yang S. Disease-specific dynamic biomarkers selected by integrating inflammatory mediators with clinical informatics in ARDS patients with severe pneumonia. Cell Biology and Toxicology. 2016; 32: 169–184.

[28] Alamgeer, Hasan UH, Uttra AM, Qasim S, Ikram J, Saleem M, et al. Phytochemicals targeting matrix metalloproteinases regulating tissue degradation in inflammation and rheumatoid arthritis. Phytomedicine. 2020; 66: 153134.

[29] Sun H, Cai S, Zhang M, Zhao J, Wei S, Luo Y, et al. MicroRNA-206 regulates vascular smooth muscle cell phenotypic switch and vascular neointimal formation. Cell Biology International. 2017; 41: 739–748.

[30] Hoesel B, Schmid JA. The complexity of NF-κB signaling in inflamma-tion and cancer. Molecular Cancer. 2013; 12: 86.

[31] Rahimifard M, Maqbool F, Moeini-Nodeh S, Niaz K, Abdollahi M, Braidy N, et al. Targeting the TLR4 signaling pathway by polyphenols: a novel therapeutic strategy for neuroinflammation. Ageing Research Reviews. 2017; 36: 11–19.