Polyphyllin II inhibits NLPR3 inflammasome activation and inflammatory response of Mycobacterium tuberculosis–infected human bronchial epithelial cells

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Guodong Cheng
Gengzhi Ye
Ying Ma
Yuqing Wang


Bronchial Epithelial Cell, Inflammatory, Mycobacterium tuberculosis, NLPR3, Polyphyllin II


Background: The bronchial infection by Mycobacterium tuberculosis (Mtb) is increasing in prevalence and severity worldwide. Despite appropriate tuberculosis treatment, most patients still develop bronchial stenosis, which often leads to disability. Polyphyllin II (PP2) is a steroidal saponin extracted from Rhizoma Paridis. In this study, we aimed to explore the effect of PP2 on the advancement of Mtb-induced bronchial infection.

Method: The effects of PP2 on cell viability were measured by using MTT and lactate dehydrogenase (LDH) kit. The mRNA and protein levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-8 were elucidated by RT-qPCR and ELISA, respectively. The expression of NLR family pyrin domain containing 3 (NLRP3) related inflammasome (NLRP3, IL-1β, and cleaved-caspase-1) and the activated degree of protein kinase B (AKT)/nuclear factor-kappa B (NF-kB; p-AKT and p-NF-κB) were detected by Western blotting.

Results: PP2 at 0, 1, 5, and 10 μM had little cytotoxicity on 16HBE cells. PP2 inhibited Mtb-induced cell proliferation and decreased LDH levels. We further found that PP2 could suppress Mtb-induced inflammatory responses and activation of NLPR3 inflammasome. Additionally, the role of PP2 in Mtb is associated with the AKT/NF-kB signaling pathway.

Conclusion: PP2 inhibited Mtb infection in bronchial epithelial cells, by inhibiting Mtb-induced inflammatory reactions and activation of NLPR3 inflammasome. These effects may be exerted by suppressing the AKT/NF-kB pathway, which will provide a prospective treatment.

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1. Parrish NM, Dick JD, Bishai WR. Mechanisms of latency in Mycobacterium tuberculosis. Trends Microbiol. 1998;6(3):107–12. 10.1016/s0966-842x(98)01216-5

2. Chai Q, Wang L, Liu CH, Ge B. New insights into the evasion of host innate immunity by Mycobacterium tuberculosis. Cell Mol Immunol. 2020;17(9):901–13. 10.1038/s41423-020-0502-z

3. MacLean E, Bigio J, Singh U, Klinton JS, Pai M. Global tuberculosis awards must do better with equity, diversity, and inclusion. Lancet. 2021;397(10270):192–3. 10.1016/S0140-6736(20)32627-1

4. Qiang L, Wang J, Zhang Y, Ge P, Chai Q, Li B, et al. Mycobacterium tuberculosis Mce2E suppresses the macrophage innate immune response and promotes epithelial cell proliferation. Cell Mol Immunol. 2019;16(4):380–91. 10.1038/s41423-018-0016-0

5. Wu Z, Zhang J, Xu F, Wang Y, Zhang J. Rapid and simple determination of polyphyllin I, II, VI, and VII in different harvest times of cultivated Paris polyphylla Smith var. yunnanensis (Franch.) Hand. Mazz by UPLC-MS/MS and FT-IR. J Nat Med. 2017;71(1):139–47. 10.1007/s11418-016-1043-8

6. Jiao Y, Xin M, Xu J, Xiang X, Li X, Jiang J, et al. Polyphyllin II induced apoptosis of NSCLC cells by inhibiting autophagy through the mTOR pathway. Pharm Biol. 2022;60(1):1781–9. 10.1080/13880209.2022.2120021

7. Chu ML, Lin PW, Liu YW, Wu SY, Lan SH, Su CL, et al. Formosanin C suppresses cancer cell proliferation and migration by impeding autophagy machinery. Kaohsiung J Med Sci. 2023;39(5):489–500. 10.1002/kjm2.12658

8. Lu Z, Yang H, Cao H, Huo C, Chen Y, Liu D, et al. Forsythoside A protects against lipopolysaccharide-induced acute lung injury through up-regulating microRNA-124. Clin Sci. 2020;134(19):2549–63. 10.1042/CS20200598

9. Yin L, Shi C, Zhang Z, Wang W, Li M. Formosanin C attenuates lipopolysaccharide-induced inflammation through nuclear factor-kappaB inhibition in macrophages. Korean J Physiol Pharmacol. 2021;25(5):395–401. 10.4196/kjpp.2021.25.5.395

10. Wang Q, Zhou X, Zhao Y, Xiao J, Lu Y, Shi Q, et al. Polyphyllin I ameliorates collagen-induced arthritis by suppressing the inflammation response in macrophages through the NF-kappaB pathway. Front Immunol. 2018;9:2091. 10.3389/fimmu.2018.02091

11. Zhang C, Li C, Jia X, Wang K, Tu Y, Wang R, et al. In vitro and in vivo anti-inflammatory effects of polyphyllin VII through downregulating MAPK and NF-kappaB pathways. Molecules. 2019;24(5):875. 10.3390/molecules24050875

12. Sun J, Zhang Q, Yang G, Li Y, Fu Y, Zheng Y, et al. The licorice flavonoid isoliquiritigenin attenuates Mycobacterium tuberculosis–induced inflammation through Notch1/NF-kappaB and MAPK signaling pathways. J Ethnopharmacol. 2022;294:115368. 10.1016/j.jep.2022.115368

13. Philips JA, Ernst JD. Tuberculosis pathogenesis and immunity. Annu Rev Pathol. 2012;7:353–84.

14. Lara-Espinosa JV, Santana-Martínez RA, Maldonado PD, Zetter M, Becerril-Villanueva E, Pérez-Sánchez G, et al. Experimental pulmonary tuberculosis in the absence of detectable brain infection induces neuroinflammation and behavioural abnormalities in male balb/c mice. Int J Mol Sci. 2020;21(24):9483. 10.3390/ijms21249483

15. Liang MY, Wang YZ, Qiao X, Lu YW, Chen MH, Li P, et al. Structural characterisation and discrimination of the aerial parts of Paris polyphylla var. yunnanensis and Paris polyphylla var. chinensis by UHPLC-QTOF-MS coupled with multivariate data analysis. Phytochem Anal. 2019;30(4):437–46. 10.1002/pca.2826

16. Man S, Chai H, Cui J, Yao J, Ma L, Gao W. Antitumor and anti-metastatic mechanisms of Rhizoma paridis saponins in Lewis mice. Environ Toxicol. 2018;33(2):149–55. 10.1002/tox.22501

17. Man S, Zhang L, Cui J, Yang L, Ma L, Gao W. Curcumin enhances the anti-cancer effects of Paris Saponin II in lung cancer cells. Cell Prolif. 2018;51(4):e12458. 10.1111/cpr.12458

18. Shifen Zhang QL, Liu L, Yang Y, Wang J. Morroniside alleviates lipopolysaccharide-induced inflammatory and oxidative stress in inflammatory bowel disease by inhibiting NLRP3 and NF-κB signaling pathways. Allergol Immunopathol (Madr). 2022;50(6):93–9. 10.15586/aei.v50i6.674

19. Junyu Li ZX, Canhui Ou Yang, Xiongjian Wu, Yun Xie, Jun Xie. Protopine alleviates lipopolysaccharide-triggered intestinal epithelial cell injury through retarding the NLRP3 and NF-κB signaling pathways to reduce inflammation and oxidative stress. Allergol Immunopathol (Madr). 2022;50(6):84–92. 10.15586/aei.v50i6.669

20. Shiliang Xie XW. CRYAB reduces cigarette smoke-induced inflammation, apoptosis, and oxidative stress by retarding PI3K/Akt and NF-κB signaling pathways in human bronchial epithelial cells. Allergol Immunopathol. 2022;50(5):23–9. 10.15586/aei.v50i5.645

21. Bickton F, Fombe C, Chisati E, Rylance J. Evidence for pulmonary rehabilitation in chronic respiratory diseases in sub-Saharan Africa: A systematic review. Int J Tuberc Lung Dis. 2020;24(10):991–9. 10.5588/ijtld.19.0526