Total alkaloids from Thesium chinense inhibit lipopolysaccharide-induced respiratory inflammation by modulating Nrf2/NF-κB/NLRP3 signaling pathway

Abstract

Inflammation plays a pivotal role in the etiology and progression of various diseases. In traditional Chinese medicine, the whole plants of Thesium chinense Turcz. and its preparations (e.g. Bairui Granules) have been employed to manage inflammatory conditions. While flavonoids were previously considered the primary anti-inflammatory components, other potentially active constituents have been largely overlooked and not thoroughly investigated. This study presents a novel finding that the total alkaloids of T. chinense (BC-Alk) are potent active substances underlying the traditional and clinical applications of T. chinense and Bairui Granules as anti-inflammatory agents. UPLC-MS/MS analysis identified the composition of BC-Alk as quinolizidine alkaloids. The anti-inflammatory efficacy of BC-Alk was evaluated using a lipopolysaccharide (LPS)-induced lung inflammation model in mice. Results demonstrated that BC-Alk significantly mitigated LPS-induced lung inflammation, attenuated the overproduction of IL-1β and the overproduction of inflammatory factors (TNF-α), and ameliorated lung tissue hyperplasia in mice in vivo. Mechanistic studies in vitro revealed that BC-Alk upregulated the expression of Nrf2 and its downstream proteins NQO1 and glutamate-cystine ligase and modifier subunit (GCLM), inhibited NF-κB phosphorylation, and suppressed NLRP3 activation. Collectively, these findings indicate that BC-Alk exerts potent inhibitory effects against lung inflammation by modulating Nrf2, NF-κB, and NLRP3 pathways. This study provides new insights into the anti-inflammatory constituents of T. chinense and Bairui Granules.

Keywords

Thesium chinense / Alkaloids / Nrf2 / NF-κB / NLRP3 / Inflammation

References

[1] Bairuicao Research Group of Anhui. Clinical observation on the treatment of acute inflammatory diseases by Thesium chinense. J Traditi Chin Med. 1972;2:38-39.

[2] Peng GC, Hao JH, Guan YQ, et al. Systematic investigation of the material basis, effectiveness and safety of Thesium chinense Turcz. and its preparation Bairui Granules against lung inflammation. Chin Med. 2024; 19(1):67. https://doi.org/10.1186/s13020-024-00940-y.

[3] Li GH, Fang KL, Yang K, et al. Thesium chinense Turcz.: an ethnomedical, phytochemical and pharmacological review. J Ethnopharmacol. 2021;273:113950. https://doi.org/10.1016/j.jep.2021.113950.

[4] Endres M, Moro MA, Nolte CH, et al. Immune pathways in etiology, acute phase, and chronic sequelae of ischemic stroke. Circ Res. 2022; 130(8):1167-1186. https://doi.org/10.1161/CIRCRESAHA.121.319994.

[5] Saviano A, Wrensch F, Ghany MG, et al. Liver disease and Coronavirus Disease 2019: from pathogenesis to clinical care. Hepatology. 2021; 74(2):1088-1100. https://doi.org/10.1002/hep.31684.

[6] Dekens DW, Eisel ULM, Gouweleeuw L, et al. Lipocalin 2 as a link between ageing, risk factor conditions and age-related brain diseases. Ageing Res Rev. 2021;70:101414. https://doi.org/10.1016/j.arr.2021.101414.

[7] Flerlage T, Boyd DF, Meliopoulos V, et al. Influenza virus and SARS-CoV-2: pathogenesis and host responses in the respiratory tract. Nat Rev Microbiol. 2021; 19(7):425-441. https://doi.org/10.1038/s41579-021-00542-7.

[8] Singh D, Long G, Cançado JED, et al. Small airway disease in chronic obstructive pulmonary disease: insights and implications for the clinician. Curr Opin Pulm Med. 2020; 26(2):162-168. https://doi.org/10.1097/MCP.0000000000000637.

[9] Bezerra FS, Lanzetti M, Nesi RT, et al. Oxidative stress and inflammation in acute and chronic lung injuries. Antioxidants (Basel). 2023; 12(3):548. https://doi.org/10.3390/antiox12030548.

[10] Yang WJ, He JX, Zhou MX, et al. An isopentenyl-substituted flavonoid norartocarpin activates Nrf2 signalling pathway and prevents oxidative insults in human lung epithelial cells. Free Radic Res. 2019; 53(3):348-358. https://doi.org/10.1080/10715762.2019.1582769.

[11] Kobayashi EH, Suzuki T, Funayama R, et al. Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription. Nat Commun. 2016;7:11624. https://doi.org/10.1038/ncomms11624.

[12] Liu T, Zhang L, Joo D, et al. NF-κB signaling in inflammation. Signal Transduct Target Ther. 2017;2:17023. https://doi.org/10.1038/sigtrans.2017.23.

[13] Mangan MSJ, Olhava EJ, Roush WR, et al. Targeting the NLRP3 inflammasome in inflammatory diseases. Nat Rev Drug Discov. 2018; 17(8):588-606. https://doi.org/10.1038/nrd.2018.97.

[14] Barnabas M, Awakan OJ, Rotimi DE, et al. Exploring redox imbalance and inflammation for asthma therapy. Mol Biol Rep. 2023; 50(9):7851-7865. https://doi.org/10.1007/s11033-023-08688-8.

[15] Tonelli C, Chio IIC, Tuveson DA.Transcriptional regulation by Nrf2. Antioxid Redox Signal. 2018; 29(17):1727-1745. https://doi.org/10.1089/ars.2017.7342.

[16] Xu LT, Wang T, Fang KL, et al. The ethanol extract of flower buds of Tussilago farfara L. attenuates cigarette smoke-induced lung inflammation through regulating NLRP3 inflammasome, Nrf2, and NF-κB. J Ethnopharmacol. 2022;283:114694. https://doi.org/10.1016/j.jep.2021.114694.

[17] Olona A, Leishman S, Anand PK. The NLRP3 inflammasome: regulation by metabolic signals. Trends Immunol. 2022; 43(12):978-989. https://doi.org/10.1016/j.it.2022.10.003.

[18] Albano GD, Gagliardo RP, Montalbano AM, et al. Overview of the mechanisms of oxidative stress: impact in inflammation of the airway diseases. Antioxidants (Basel). 2022; 11(11):2237. https://doi.org/10.3390/antiox11112237.

[19] Jeong YJ, Kim CU, Lee KS, et al. Pseudomonas stutzeri PM101005 inhaled with atmospheric particulate matter induces lung damage through inflammatory responses. Environ Pollut. 2023;317:120741. https://doi.org/10.1016/j.envpol.2022.120741.

[20] Rosenwasser Y, Berger I, Loewy ZG. Therapeutic approaches for chronic obstructive pulmonary disease (COPD) exacerbations. Pathogens. 2022; 11(12):1513. https://doi.org/10.3390/pathogens11121513.

[21] Huang K, Zhang P, Zhang Z, et al. Traditional Chinese medicine (TCM) in the treatment of COVID-19 and other viral infections: efficacies and mechanisms. Pharmacol Ther. 2021;225:107843. https://doi.org/10.1016/j.pharmthera.2021.107843.

[22] He YQ, Zhou CC, Yu LY, et al. Natural product derived phytochemicals in managing acute lung injury by multiple mechanisms. Pharmacol Res. 2021;163:105224. https://doi.org/10.1016/j.phrs.2020.105224.

[23] Yang M, Wang Y, Fan Z, et al.Chemical constituents and anti-inflammatory activity of the total alkaloid extract from Melodinus cochinchinensis (Lour.) Merr. and its inhibition of the NF-κB and MAPK signaling pathways. Phytomedicine. 2021;91:153684. https://doi.org/10.1016/j.phymed.2021.153684.

[24] Bai R, Yao C, Zhong Z, et al. Discovery of natural anti-inflammatory alkaloids: potential leads for the drug discovery for the treatment of inflammation. Eur J Med Chem. 2021;213:113165. https://doi.org/10.1016/j.ejmech.2021.113165.

[25] Sabzehzari M, Zeinali M, Naghavi MR. Alternative sources and metabolic engineering of Taxol: advances and future perspectives. Biotechnol Adv. 2020;43:107569. https://doi.org/10.1016/j.biotechadv.2020.107569.

[26] Dhyani P, Quispe C, Sharma E, et al. Anticancer potential of alkaloids: a key emphasis to colchicine, vinblastine, vincristine, vindesine, vinorelbine and vincamine. Cancer Cell Int. 2022; 22(1):206. https://doi.org/10.1186/s12935-022-02624-9.

[27] Parveen Z, Deng Y, Saeed MK, et al. Antiinflammatory and analgesic activities of Thesium chinense Turcz. extracts and its major flavonoids, kaempferol and kaempferol-3-O-glucoside. Yakugaku Zasshi. 2007; 127(8):1275-1279. https://doi.org/10.1248/yakushi.127.1275.

[28] Wang Z, Li SS. Isolation and identification of alkaloids from Thesium chinense Turcz. Chin J Med Chem. 2006; 16(5):306-308. https://doi.org/10.1007/978-3-642-40047-6_3.

[29] Lu Y, Xu D, Liu J, et al. Protective effect of sophocarpine on lipopolysaccharide-induced acute lung injury in mice. Int Immunopharmacol. 2019; 70:180-186. https://doi.org/10.1016/j.intimp.2019.02.020.

[30] Cuadrado A, Rojo AI, Wells G, et al. Therapeutic targeting of the NRF2 and KEAP1 partnership in chronic diseases. Nat Rev Drug Discov. 2019; 18(4):295-317. https://doi.org/10.1038/s41573-018-0008-x.

[31] Kaulmann A, Bohn T. Carotenoids, inflammation, and oxidative stress-implications of cellular signaling pathways and relation to chronic disease prevention. Nutr Res. 2014; 34(11):907-929. https://doi.org/10.1016/j.nutres.2014.07.010.

[32] Reuter S, Gupta SC, Chaturvedi MM, et al. Oxidative stress, inflammation, and cancer: how are they linked? Free Radic Biol Med. 2010; 49(11):1603-1616. https://doi.org/10.1016/j.freeradbiomed.2010.09.006.

[33] He F, Antonucci L, Karin M. NRF2 as a regulator of cell metabolism and inflammation in cancer. Carcinogenesis. 2020; 41(4):405-416. https://doi.org/10.1093/carcin/bgaa039.

[34] Ahmed SM, Luo L, Namani A, et al. Nrf2 signaling pathway: pivotal roles in inflammation. Biochim Biophys Acta Mol Basis Dis. 2017; 1863(2):585-597. https://doi.org/10.1016/j.bbadis.2016.11.005.

[35] Wu J, Chen P, Ju L, et al. Corydalis saxicola Bunting total alkaloids ameliorate diet-induced non-alcoholic steatohepatitis by regulating hepatic PI3K/Akt and TLR4/NF-κB pathways in mice. Biomed Pharmacother. 2022;151:113132. https://doi.org/10.1016/j.biopha.2022.113132.

[36] Wu Y, Liu Y, Zhang L, et al. Aconiti Lateralis Radix Praeparata total alkaloids exert anti-RA effects by regulating NF-κB and JAK/STAT signaling pathways and promoting apoptosis. Front Pharmacol. 2022;13:980229. https://doi.org/10.3389/fphar.2022.980229.

[37] Cui YR, Qu F, Zhong WJ, et al. Beneficial effects of aloperine on inflammation and oxidative stress by suppressing necroptosis in lipopolysaccharide-induced acute lung injury mouse model. Phytomedicine. 2022;100:154074. https://doi.org/10.1016/j.phymed.2022.154074.

[38] Jo EK, Kim JK, Shin DM, et al. Molecular mechanisms regulating NLRP3 inflammasome activation. Cell Mol Immunol. 2016; 13(2):148-159. https://doi.org/10.1038/cmi.2015.95.

[39] Liu X, Zhang X, Ding Y, et al.Nuclear factor E2-related factor-2 negatively regulates NLRP3 inflammasome activity by inhibiting reactive oxygen species-induced NLRP3 priming. Antioxid Redox Signal. 2017; 26(1):28-43. https://doi.org/10.1089/ars.2015.6615.

[40] Donovan C, Liu G, Shen S, et al. The role of the microbiome and the NLRP3 inflammasome in the gut and lung. J Leukoc Biol. 2020; 108(3):925-935. https://doi.org/10.1002/JLB.3MR0720-472RR.

[41] Huang Y, Xu W, Zhou R. NLRP3 inflammasome activation and cell death. Cell Mol Immunol. 2021; 18(9):2114-2127. https://doi.org/10.1038/s41423-021-00740-6.

[42] Kelley N, Jeltema D, Duan Y, et al. The NLRP3 inflammasome: an overview of mechanisms of activation and regulation. Int J Mol Sci. 2019; 20(13):3328. https://doi.org/10.3390/ijms20133328.

[43] Bai Y, Mu Q, Bao X, et al. Targeting NLRP3 inflammasome in the treatment of diabetes and diabetic complications: role of natural compounds from herbal medicine. Aging Dis. 2021; 12(7):1587-1604. https://doi.org/10.14336/AD.2021.0318.

[44] Bagherniya M, Khedmatgozar H, Fakheran O, et al. Medicinal plants and bioactive natural products as inhibitors of NLRP3 inflammasome. Phytother Res. 2021; 35(9):4804-4833. https://doi.org/10.1002/ptr.7118.