Abstract
Interstitial cystitis (IC) is a chronic bladder inflammation. Inhibition of prostaglandin G/H synthase 2 (PTGS2) is the most common method for controlling inflammation-related diseases. This study aimed to analyze the effects of hispidulin on the PTGS2 and NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammation in experimental IC models. A binding activity between hispidulin and PTGS2 was measured using molecular docking. Human urothelial cells (SV-HUC-1) were stimulated by 2 ng/mL of interleukin (IL)-1β for 24 h and cultured in a medium with different concentrations of hispidulin (2.5, 5, 10, 20 µM) for 24 h to observe the expressions of PTGS2 and NLRP3 protein. Cells overexpressing PTGS2 were established by PTGS2 cDNA transfection. In the IL-1β-treated cells, the NLRP3 inflammasome was measured after 20 µM hispidulin treatment. In rats, animals were performed with three injections of 40 mg/kg cyclophosphamide (CYP) and orally treated with 50 mg/kg/day hispidulin or ibuprofen for 3 days. The bladder pain was measured using Von Frey filaments, and the bladder pathology was observed using hematoxylin and eosin (H&E) staining. The expressions of PTGS2 and NLRP3 inflammasome were also observed in the bladder tissues. A good binding activity was found between hispidulin and PTGS2 (score = − 8.9 kcal/mol). The levels of PTGS2 and NLRP3 inflammasome were decreased with the hispidulin dose increase in the IL-1β-treated cells (p < 0.05). Cells overexpressing PTGS2 weakened the protective effects of hispidulin in the IL-1β-treated cells (p < 0.01). In the CYP-treated rats, hispidulin treatment improved the bladder pain through decreasing the nociceptive score (p < 0.01) and suppressed the bladder inflammation through suppressing the expressions of PTGS2 and NLRP3 inflammasome in bladder tissues (p < 0.01). Additionally, the results of ibuprofen treatment were similar to the effects of hispidulin in the CYP-treated rats. This study demonstrates that hispidulin may be a new alternative drug for the IC treatment that binds PTGS2 to perform its functions.
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The authors declare that all data were generated in-house and that no paper mill was used.
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The datasets generated during the current study are available from the corresponding author.
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S.L.L. and S.H.L. participated in the study design and the data collection and analysis. Y.P.D. and K.Q. coordinated experimental work and data collection. Y.Z. partook the manuscript preparation. S.L.L. wrote the manuscript. J.Y.Y. participated in the study design and revised the manuscript. The final manuscript was read and agreed with all authors. The authors declare that all data were generated in-house and that no paper mill was used.
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All experiment processes were agreed with the Ethics Committee of Yantai Affiliated Hospital of Binzhou Medical University (Approval No. 20230608054). All experimental procedures were performed in accordance to the ARRIVE guidelines for the reporting of animal experiments.
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Liu, S., Li, S., Dong, Y. et al. Hispidulin targets PTGS2 to improve cyclophosphamide-induced cystitis by suppressing NLRP3 inflammasome. Naunyn-Schmiedeberg's Arch Pharmacol 397, 5819–5830 (2024). https://doi.org/10.1007/s00210-024-02987-y
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DOI: https://doi.org/10.1007/s00210-024-02987-y