The Mechanism of Notopterol Alleviating LPS-induced Endometritis by Inhibiting the TLR4/NF-κB Signaling Pathway
,
Abstract
This study aims to investigate the role of notopterol in alleviating endometritis induced by lipopolysaccharide (LPS) and to explore its underlying mechanisms.Human endometrial epithelial cells (hEECs) were treated with LPS to establish an in vitro model of endometritis, and the cells were divided into five groups: control, LPS, LPS+notopterol(15 mol/L), LPS+notopterol(305 mol/L) and LPS+notopterol(45 mol/L) groups. The expression levels of inflammatory factors were determined by Enzyme-Linked ImmunoSorbent Assay (ELISA). Apoptosis was detected by TdT-mediated dUTP Nick-End Labeling (TUNEL) method. Cell viability was determined by Cell Counting Kit-8 (CCK-8) test. Western blot was used to detect the expression levels of nuclear factor κB(NF-κB) p65, NF-κB inhibitor (IκBα), p-NF-κB p65 and p-IκBα.
Following LPS treatment, cytokine levels significantly increased compared to the control group.; moreover, cell proliferation decreased, apoptosis increased, and the expression level of p-NF-κB p65 was increased. Subsequently, the LPS-treated hEECs were exposed to notopterygium. Compared to the LPS group.
Treatment with LPS + notopterol resulted in a dose-dependent reduction in inflammatory cytokines, increased cell proliferation, and a significant reduction in apoptosis. Furthermore, the expression levels of p-NF-κB p65 and p-IκBα were downregulated.
These findings suggest that notopterol alleviates LPS-induced endometritis by inhibiting the TLR4/NF-κB signaling pathway.
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10. Schöniger S, Gräfe H, Schoon HA. Expression of Toll-like receptors 2, 4 and 6 in different cell populations of the equine endometrium. Veterinary Immunol Immunopathol. 2017;185:7-13.
11. Aboussahoud W, Aflatoonian R, Bruce C, et al. Expression and function of Toll-like receptors in human endometrial epithelial cell lines. J Reproductive Immunol. 2010;84(1):41-51.
12. Huang Y, Xu W, Zhou R. NLRP3 inflammasome activation and cell death. Cell Mol Immunol. 2021;18(9):2114-2127.
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14. Ruan Y, Jin X, Ji H. Water extract of Notopterygium incisum alleviates cold allodynia in neuropathic pain by regulation of TRPA1. J Ethnopharmacol. 2023;305:116065.
15. Jiang XW, Liu WW, Wu YT. Notopterygium incisum extract (NRE) rescues cognitive deficits in APP/PS1 Alzhneimer's disease mice by attenuating amyloid-beta, tau, and neuroinflammation pathology. J Ethnopharmacol. 2020;249:112433.
16. Bi JP, Li P, Xu XX, Wang T, Li F. Anti-rheumatoid arthritic effect of volatile components in notopterygium incisum in rats via anti-inflammatory and anti-angiogenic activities. Chinese J Nat Med. 2018;16(12):926-935.
17. Kim D, Dasgupta S, Ma X, Park JM, Wei HT, Li X, et al. Rapid discrimination of Notopterygium incisum and Notopterygium franchetii based on characteristic compound profiles detected by UHPLC-QTOF-MS/MS coupled with multivariate analysis. Sci Adv. 2020;31(3):355-65.
18. Zhou W, Van Sinderen M, Rainczuk K, Menkhorst E, Sorby K, Osianlis T, et al. Dysregulated miR-124-3p in endometrial epithelial cells reduces endometrial receptivity by altering polarity and adhesion. Proc Natl Acad Sci U S A. 2024;121(41):e2401071121.
19. Qian X, Wang Y, Li X, Li Y, Li L. TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation. Immun Inflamm Dis. 2023;11(10):e970.
20. Zhao R, Wang J, Zhang X, Chen Y. MiR-643 inhibits lipopolysaccharide-induced endometritis progression by targeting TRAF6. Cell Biol Int. 20 20;44(4):1059-1067.
21. Choksuchat C, Zhao S, Deutch TD, Kimble TD, Archer DF. Effects of progesterone, levonorgestrel and medroxyprogesterone acetate on apoptosis in human endometrial endothelial cells. Contraception. 2009;79(2):139-45.
22. Wang L, Li S, Luo H, Lu Q, Yu S. PCSK9 promotes the progression and metastasis of colon cancer cells through regulation of EMT and PI3K/AKT signaling in tumor cells and phenotypic polarization of macrophages. J Exp Clin Cancer Res. 2022;41(1):303.
23. Li XQ, Kang R, Huo JC, Xie YH, Wang SW, Cao W. Wound-healing Activity of Zanthoxylum bungeanum Maxim Seed Oil on Experimentally Burned Rats. Pharmacognosy Magazine. 2017;13(51):363-71.
24. Singh N, Sethi A. Endometritis - Diagnosis,Treatment and its impact on fertility - A Scoping Review. JBRA Assist Reprod. 2022;26(3):538-46.
25. Peng J, Guo F, Liu H, Gu Z, Zeng Z, Li M, et al. Correlation between hysteroscopy findings in patients with hydrosalpinx and chronic endometritis. Int J Gynaecol Obstet. 2022;157(2):471-5.
26. Shen Y, Liu B, Mao W, Gao R, Feng S, Qian Y, et al. PGE(2) downregulates LPS-induced inflammatory responses via the TLR4-NF-κB signaling pathway in bovine endometrial epithelial cells. Prostaglandins Leukot Essent Fatty Acids. 2018;129:25-31.
27. Shi J, Zhao Y, Wang Y. Inflammatory caspases are innate immune receptors for intracellular LPS. Nature. 2014;514(7521):187-92.
28. Zhou J, Shi P, Ma R, Xie X, Zhao L, Wang J. Notopterol Inhibits the NF-κB Pathway and Activates the PI3K/Akt/Nrf2 Pathway in Periodontal Tissue. J Immunol. 2023;211(10):1516-25.
29. Chen KT, Yeh CT, Yadav VK. Notopterol mitigates IL-1β-triggered pyroptosis by blocking NLRP3 inflammasome via the JAK2/NF-kB/hsa-miR-4282 route in osteoarthritis. Heliyon. 2024;10(6):e28094.
30. Xu K, Jiang S, Zhou Y, Zhang Y, Xia B, Xu X, et al. Discrimination of the seeds of Notopterygium incisum and Notopterygium franchetii by validated HPLC-DAD-ESI-MS method and principal component analysis. Pharm Biomed Anal. 2011;56(5):1089-1093.
31. Haque MS, Kim B, You MJ. Glycosides from the methanol extract of Notopterygium incisum. Planta Med. 2011;77(17):1939-43.
2. Li J, Zhao Y, Gao Y, Zhu Y, Holyoak GR, Zeng S. Treatments for Endometritis in Mares Caused by Streptococcus equi Subspecies zooepidemicus: A Structured Literature Review. J Equine Vet Sci. 2021;102(12):103430.
3. Yin B, Umar T, Ma X, Chen Y, Umar Z, Umer S, Deng G. Andrograpanin mitigates lipopolysaccharides induced endometritis via TLR4/NF-κB pathway. Reprod Biol. 2022;22(1):100606.
4. Kitaya K, Takeuchi T, Mizuta S, Matsubayashi H, Ishikawa T. Endometritis: new time, new concepts. Fertil Ster. 2018;110(3):344-50.
5. Mackeen AD, Packard RE, Ota E, Speer L. Antibiotic regimens for postpartum endometritis. Cochrane Database Syst Rev. 2015;2015(2):Cd001067.
6. Cicinelli E, Cicinelli R, Vitagliano A. Antibiotic therapy for chronic endometritis and its reproductive implications: a step forward, with some uncertainties. Fertility and sterility. 2021;115(6):1445-6.
7. Wang L, Yang JW, Lin LT, et al. Acupuncture Attenuates Inflammation in Microglia of Vascular Dementia Rats by Inhibiting miR-93-Mediated TLR4/MyD88/NF-κB Signaling Pathway. Oxidative medicine and cellular longevity. 2020;2020:8253904.
8. Wu L, Du L, Ju Q. Silencing TLR4/MyD88/NF-κB Signaling Pathway Alleviated Inflammation of Corneal Epithelial Cells Infected by ISE. Inflammation. 2021;44(2):633-644.
9. Hu N, Wang C, Dai X. Phillygenin inhibits LPS-induced activation and inflammation of LX2 cells by TLR4/MyD88/NF-κB signaling pathway. J Ethnopharmacol. 2020;248:112361.
10. Schöniger S, Gräfe H, Schoon HA. Expression of Toll-like receptors 2, 4 and 6 in different cell populations of the equine endometrium. Veterinary Immunol Immunopathol. 2017;185:7-13.
11. Aboussahoud W, Aflatoonian R, Bruce C, et al. Expression and function of Toll-like receptors in human endometrial epithelial cell lines. J Reproductive Immunol. 2010;84(1):41-51.
12. Huang Y, Xu W, Zhou R. NLRP3 inflammasome activation and cell death. Cell Mol Immunol. 2021;18(9):2114-2127.
13. Azietaku JT, Ma H, Yu XA. A review of the ethnopharmacology, phytochemistry and pharmacology of Notopterygium incisum. J ethnopharmacol. 2017;202:241-55.
14. Ruan Y, Jin X, Ji H. Water extract of Notopterygium incisum alleviates cold allodynia in neuropathic pain by regulation of TRPA1. J Ethnopharmacol. 2023;305:116065.
15. Jiang XW, Liu WW, Wu YT. Notopterygium incisum extract (NRE) rescues cognitive deficits in APP/PS1 Alzhneimer's disease mice by attenuating amyloid-beta, tau, and neuroinflammation pathology. J Ethnopharmacol. 2020;249:112433.
16. Bi JP, Li P, Xu XX, Wang T, Li F. Anti-rheumatoid arthritic effect of volatile components in notopterygium incisum in rats via anti-inflammatory and anti-angiogenic activities. Chinese J Nat Med. 2018;16(12):926-935.
17. Kim D, Dasgupta S, Ma X, Park JM, Wei HT, Li X, et al. Rapid discrimination of Notopterygium incisum and Notopterygium franchetii based on characteristic compound profiles detected by UHPLC-QTOF-MS/MS coupled with multivariate analysis. Sci Adv. 2020;31(3):355-65.
18. Zhou W, Van Sinderen M, Rainczuk K, Menkhorst E, Sorby K, Osianlis T, et al. Dysregulated miR-124-3p in endometrial epithelial cells reduces endometrial receptivity by altering polarity and adhesion. Proc Natl Acad Sci U S A. 2024;121(41):e2401071121.
19. Qian X, Wang Y, Li X, Li Y, Li L. TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation. Immun Inflamm Dis. 2023;11(10):e970.
20. Zhao R, Wang J, Zhang X, Chen Y. MiR-643 inhibits lipopolysaccharide-induced endometritis progression by targeting TRAF6. Cell Biol Int. 20 20;44(4):1059-1067.
21. Choksuchat C, Zhao S, Deutch TD, Kimble TD, Archer DF. Effects of progesterone, levonorgestrel and medroxyprogesterone acetate on apoptosis in human endometrial endothelial cells. Contraception. 2009;79(2):139-45.
22. Wang L, Li S, Luo H, Lu Q, Yu S. PCSK9 promotes the progression and metastasis of colon cancer cells through regulation of EMT and PI3K/AKT signaling in tumor cells and phenotypic polarization of macrophages. J Exp Clin Cancer Res. 2022;41(1):303.
23. Li XQ, Kang R, Huo JC, Xie YH, Wang SW, Cao W. Wound-healing Activity of Zanthoxylum bungeanum Maxim Seed Oil on Experimentally Burned Rats. Pharmacognosy Magazine. 2017;13(51):363-71.
24. Singh N, Sethi A. Endometritis - Diagnosis,Treatment and its impact on fertility - A Scoping Review. JBRA Assist Reprod. 2022;26(3):538-46.
25. Peng J, Guo F, Liu H, Gu Z, Zeng Z, Li M, et al. Correlation between hysteroscopy findings in patients with hydrosalpinx and chronic endometritis. Int J Gynaecol Obstet. 2022;157(2):471-5.
26. Shen Y, Liu B, Mao W, Gao R, Feng S, Qian Y, et al. PGE(2) downregulates LPS-induced inflammatory responses via the TLR4-NF-κB signaling pathway in bovine endometrial epithelial cells. Prostaglandins Leukot Essent Fatty Acids. 2018;129:25-31.
27. Shi J, Zhao Y, Wang Y. Inflammatory caspases are innate immune receptors for intracellular LPS. Nature. 2014;514(7521):187-92.
28. Zhou J, Shi P, Ma R, Xie X, Zhao L, Wang J. Notopterol Inhibits the NF-κB Pathway and Activates the PI3K/Akt/Nrf2 Pathway in Periodontal Tissue. J Immunol. 2023;211(10):1516-25.
29. Chen KT, Yeh CT, Yadav VK. Notopterol mitigates IL-1β-triggered pyroptosis by blocking NLRP3 inflammasome via the JAK2/NF-kB/hsa-miR-4282 route in osteoarthritis. Heliyon. 2024;10(6):e28094.
30. Xu K, Jiang S, Zhou Y, Zhang Y, Xia B, Xu X, et al. Discrimination of the seeds of Notopterygium incisum and Notopterygium franchetii by validated HPLC-DAD-ESI-MS method and principal component analysis. Pharm Biomed Anal. 2011;56(5):1089-1093.
31. Haque MS, Kim B, You MJ. Glycosides from the methanol extract of Notopterygium incisum. Planta Med. 2011;77(17):1939-43.
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| Endometritis Inflammation Lipopolysaccharide Notopterol | ||
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How to Cite
1.
Xu Z, Zhang Y, Pang J, Chen X, Chen Y, Chen Y, Wang Y. The Mechanism of Notopterol Alleviating LPS-induced Endometritis by Inhibiting the TLR4/NF-κB Signaling Pathway. Iran J Allergy Asthma Immunol. 2025;:1-9.
