Abstract
The aim of this study was to investigate the alleviating effect of wogonin on intracerebral hemorrhage (ICH) and its mechanism. The hemin-treated PC-12 cells were constructed to mimic ICH in vitro. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis was used for cell viability measurement and flow cytometry was for pyroptosis detection. Enzyme-linked immunosorbent assay (ELISA) assay and western blot were used to detect the protein levels of pyroptosis-related proteins. The modification level of N6-methyladenosine (m6A) methylation was detected by quantitative real-time polymerase chain reaction (qRT-PCR) combined with m6A dot blot assays. Molecular docking experiments analyzed the binding of wogonin and METTL14 protein. The correlation between METTL14 and NLRP3 was confirmed by bioinformatics analysis and dual luciferase reporter gene detection. ICH was induced in mice injected with collagenase into the basal ganglia, and the neurobehavioral damage was evaluated. Triphenyltetrazolium chloride monohydrate (TTC) staining and neurological scores were used to assess brain damage in mice. The results demonstrated that wogonin alleviated neuronal cell pyroptosis, and was molecularly docked with METTL14. Overexpression of METTL14 partly reversed the protecting effects of wogonin on brain in vitro and in vivo. Furthermore, NLRP3 was methylated by METTL14. Taken together, wogonin inhibits neuronal pyroptosis and thus treats IHC by inhibiting METTL14 and its methylated NLRP3.
Similar content being viewed by others
References
Kn, S. (2022). Spontaneous intracerebral hemorrhage. The New England Journal of Medicine, 387(17), 1589–1596. https://doi.org/10.1056/NEJMra2201449
Greenberg, S. M., Ziai, W. C., Cordonnier, C., Dowlatshahi, D., Francis, B., Goldstein, J. N., Hemphill, R. J. C., Johnson, R., Keigher, K. M., Mack, W. J., Mocco, J., Newton, E. J., Ruff, I. M., Sansing, L. H., Schulman, S., Selim, M. H., Sheth, K. N., Sprigg, N., Sunnerhagen, K. S., et al. (2022). 2022 guideline for the management of patients with spontaneous intracerebral hemorrhage: A guideline from the American Heart Association/American Stroke Association. Stroke, 53(7), 101161S–101407S. https://doi.org/10.1161/STR.0000000000000407
Zhou, J. F., Xiong, Y., Kang, X., Pan, Z., Zhu, Q., Goldbrunner, R., Stavrinou, L., Lin, S., Hu, W., Zheng, F., & Stavrinou, P. (2022). Application of stem cells and exosomes in the treatment of intracerebral hemorrhage: An update. Stem Cell Research & Therapy, 13(1), 281. https://doi.org/10.1186/s13287-022-02965-2
Yu, Z., Zhang, L., Zhang, G., Xia, K., Yang, Q., Huang, T., & Fan, D. (2022). Lipids, apolipoproteins, statins, and intracerebral hemorrhage: A mendelian randomization study. Annals of Neurology, 92(3), 390–399. https://doi.org/10.1002/ana.26426
Duan, T., Li, L., Yu, Y., Li, T., Han, R., Sun, X., Cui, Y., Liu, T., Wang, X., Wang, Y., Fan, X., Liu, Y., & Zhang, H. (2022). Traditional Chinese medicine use in the pathophysiological processes of intracerebral hemorrhage and comparison with conventional therapy. Pharmacological Research, 179, 106200. https://doi.org/10.1016/j.phrs.2022.106200
Baradaran, R. V., Askari, V. R., & Hosseinzadeh, H. (2021). Promising influences of scutellaria baicalensis and its two active constituents, baicalin, and baicalein, against metabolic syndrome: A review. Phytotherapy Research, 35(7), 3558–3574. https://doi.org/10.1002/ptr.7046
Banik, K., Khatoon, E., Harsha, C., Rana, V., Parama, D., Thakur, K. K., Bishayee, A., & Kunnumakkara, A. B. (2022). Wogonin and its analogs for the prevention and treatment of cancer: A systematic review. Phytotherapy Research, 36(5), 1854–1883. https://doi.org/10.1002/ptr.7386
Khan, N. M., Haseeb, A., Ansari, M. Y., Devarapalli, P., Haynie, S., & Haqqi, T. M. (2017). Wogonin, a plant derived small molecule, exerts potent anti-inflammatory and chondroprotective effects through the activation of ros/erk/nrf2 signaling pathways in human osteoarthritis chondrocytes. Free Radical Biology & Medicine, 106, 288–301. https://doi.org/10.1016/j.freeradbiomed.2017.02.041
Feng, Y., Ju, Y., Yan, Z., Ji, M., Yang, M., Wu, Q., Wang, L., & Sun, G. (2022). Protective role of wogonin following traumatic brain injury by reducing oxidative stress and apoptosis via the pi3k/nrf2/ho-1 pathway. International Journal of Molecular Medicine, 49(4). https://doi.org/10.3892/ijmm.2022.5109
Wang, L., Li, C., Sreeharsha, N., Mishra, A., Shrotriya, V., & Sharma, A. (2020). Neuroprotective effect of wogonin on rat’s brain exposed to gamma irradiation. Journal of Photochemistry and Photobiology. B, Biology, 204, 111775. https://doi.org/10.1016/j.jphotobiol.2020.111775
Zhuang, J., Peng, Y., Gu, C., Chen, H., Lin, Z., Zhou, H., Wu, X., Li, J., Yu, X., Cao, Y., Zeng, H., Fu, X., Xu, C., Huang, P., Cao, S., Wang, C., Yan, F., & Chen, G. (2021). Wogonin accelerates hematoma clearance and improves neurological outcome via the ppar-gamma pathway after intracerebral hemorrhage. Translational Stroke Research, 12(4), 660–675. https://doi.org/10.1007/s12975-020-00842-9
Yu, P., Zhang, X., Liu, N., Tang, L., Peng, C., & Chen, X. (2021). Pyroptosis: Mechanisms and diseases. Signal Transduction and Targeted Therapy, 6(1), 128. https://doi.org/10.1038/s41392-021-00507-5
Gu, L., Sun, M., Li, R., Zhang, X., Tao, Y., Yuan, Y., Luo, X., & Xie, Z. (2022). Didymin suppresses microglia pyroptosis and neuroinflammation through the asc/caspase-1/gsdmd pathway following experimental intracerebral hemorrhage. Frontiers in Immunology, 13, 810582. https://doi.org/10.3389/fimmu.2022.810582
Song, D., Yeh, C. T., Wang, J., & Guo, F. (2022). Perspectives on the mechanism of pyroptosis after intracerebral hemorrhage. Frontiers in Immunology, 13, 989503. https://doi.org/10.3389/fimmu.2022.989503
Gu, L., Sun, M., Li, R., Tao, Y., Luo, X., Zhang, X., Yuan, Y., & Xie, Z. (2022). Microglial pyroptosis: Therapeutic target in secondary brain injury following intracerebral hemorrhage. Frontiers in Cellular Neuroscience, 16, 971469. https://doi.org/10.3389/fncel.2022.971469
Ding, Z., Zhong, Z., Wang, J., Zhang, R., Shao, J., Li, Y., Wu, G., Tu, H., Yuan, W., Sun, H., & Wang, Q. (2022). Inhibition of dectin-1 alleviates neuroinflammatory injury by attenuating nlrp3 inflammasome-mediated pyroptosis after intracerebral hemorrhage in mice: Preliminary study results. Journal of Inflammation Research, 15, 5917–5933. https://doi.org/10.2147/JIR.S384020
Tian, M., Mao, L., & Zhang, L. (2022). Crosstalk among n6-methyladenosine modification and rnas in central nervous system injuries. Frontiers in Cellular Neuroscience, 16, 1013450. https://doi.org/10.3389/fncel.2022.1013450
Zhang, H., Shi, X., Huang, T., Zhao, X., Chen, W., Gu, N., & Zhang, R. (2020). Dynamic landscape and evolution of m6a methylation in human. Nucleic Acids Research, 48(11), 6251–6264. https://doi.org/10.1093/nar/gkaa347
Zhang, C., & Liu, N. (2022). N6-methyladenosine (m6a) modification in gynecological malignancies. Journal of Cellular Physiology, 237(9), 3465–3479. https://doi.org/10.1002/jcp.30828
Duan, L., Zhang, Y., Yang, Y., Su, S., Zhou, L., Lo, P. C., Cai, J., Qiao, Y., Li, M., Huang, S., Wang, H., Mo, Y., & Wang, Q. (2021). Baicalin inhibits ferroptosis in intracerebral hemorrhage. Frontiers in Pharmacology, 12, 629379. https://doi.org/10.3389/fphar.2021.629379
Chen, C. C., Hung, T. H., Wang, Y. H., Lin, C. W., Wang, P. Y., Lee, C. Y., & Chen, S. F. (2012). Wogonin improves histological and functional outcomes, and reduces activation of tlr4/nf-κb signaling after experimental traumatic brain injury. PLoS One, 7(1), e30294. https://doi.org/10.1371/journal.pone.0030294
Gan, H., Zhang, L., Chen, H., Xiao, H., Wang, L., Zhai, X., Jiang, N., Liang, P., Zheng, S., & Zhao, J. (2021). The pivotal role of the nlrc4 inflammasome in neuroinflammation after intracerebral hemorrhage in rats. Experimental & Molecular Medicine, 53(11), 1807–1818. https://doi.org/10.1038/s12276-021-00702-y
Jin, P., Qi, D., Cui, Y., Lenahan, C., Zhang, J. H., Tao, X., Deng, S., & Tang, J. (2022). Aprepitant attenuates nlrc4-dependent neuronal pyroptosis via nk1r/pkcδ pathway in a mouse model of intracerebral hemorrhage. Journal of Neuroinflammation, 19(1), 1–198. https://doi.org/10.1186/s12974-022-02558-z
Pi, Z., Liu, J., Xiao, H., & Hu, Z. (2021). L-3-n-butylphthalide promotes restoration after an experimental animal model of intracerebral hemorrhage. International Journal of Medical Sciences, 18(12), 2607–2614. https://doi.org/10.7150/ijms.60342
Yan, J., Xu, W., Lenahan, C., Huang, L., Wen, J., Li, G., Hu, X., Zheng, W., Zhang, J. H., & Tang, J. (2021). Ccr5 activation promotes nlrp1-dependent neuronal pyroptosis via ccr5/pka/creb pathway after intracerebral hemorrhage. Stroke, 52(12), 4021–4032. https://doi.org/10.1161/STROKEAHA.120.033285
Yang, C., Wang, T., Chen, J., He, J., Li, Y., Chen, C., Lu, G., & Chen, W. (2021). Traditional Chinese medicine formulas alleviate acute pancreatitis: Pharmacological activities and mechanisms. Pancreas, 50(10), 1348–1356. https://doi.org/10.1097/MPA.0000000000001931
Al-Kawaz, M., Cho, S. M., Gottesman, R. F., Suarez, J. I., & Rivera-Lara, L. (2022). Impact of cerebral autoregulation monitoring in cerebrovascular disease: A systematic review. Neurocritical Care, 36(3), 1053–1070. https://doi.org/10.1007/s12028-022-01484-5
Lin, W., Hou, J., Han, T., Zheng, L., Liang, H., & Zhou, X. (2022). Efficacy and safety of traditional Chinese medicine for intracranial hemorrhage by promoting blood circulation and removing blood stasis: A systematic review and meta-analysis of randomized controlled trials. Frontiers in Pharmacology, 13, 942657. https://doi.org/10.3389/fphar.2022.942657
Shin, J. W., Kang, H. C., Shim, J., & Sohn, N. W. (2012). Scutellaria baicalensis attenuates blood-brain barrier disruption after intracerebral hemorrhage in rats. The American Journal of Chinese Medicine, 40(1), 85–96. https://doi.org/10.1142/S0192415X12500073
Zhou, Q., Jin, Y., Jia, Q., Zhang, Y., Li, L., Liu, P., & Liu, Y. (2014). Baicalin attenuates brain edema in a rat model of intracerebral hemorrhage. Inflammation, 37(1), 107–115. https://doi.org/10.1007/s10753-013-9717-9
Zhao, X., Qiao, D., Guan, D., Wang, K., & Cui, Y. (2022). Chrysophanol ameliorates hemin-induced oxidative stress and endoplasmic reticulum stress by regulating microrna-320-5p/wnt3a pathway in ht22 cells. Oxidative Medicine and Cellular Longevity, 2022, 9399658. https://doi.org/10.1155/2022/9399658
Jin, Z. L., Gao, W. Y., Liao, S. J., Yu, T., Shi, Q., Yu, S. Z., & Cai, Y. F. (2021). Paeonol inhibits the progression of intracerebral haemorrhage by mediating the hotair/upf1/acsl4 axis. ASN Neuro, 13, 523085737. https://doi.org/10.1177/17590914211010647
Zhang, Y., Lu, P., Qin, H., Zhang, Y., Sun, X., Song, X., Liu, J., Peng, H., Liu, Y., Nwafor, E. O., Li, J., & Liu, Z. (2021). Traditional Chinese medicine combined with pulmonary drug delivery system and idiopathic pulmonary fibrosis: Rationale and therapeutic potential. Biomedicine & Pharmacotherapy, 133, 111072. https://doi.org/10.1016/j.biopha.2020.111072
Jiang, H., Yao, Q., An, Y., Fan, L., Wang, J., & Li, H. (2022). Baicalin suppresses the progression of Type 2 diabetes-induced liver tumor through regulating METTL3/m6A/HKDC1 axis and downstream p-JAK2/STAT1/clevaged Capase3 pathway. Phytomedicine, 94, 153823. https://doi.org/10.1016/j.phymed.2021.153823
Zhang, L., Wang, X., Che, W., Yi, Y., Zhou, S., & Feng, Y. (2022). Methyltransferase-like 3 silenced inhibited the ferroptosis development via regulating the glutathione peroxidase 4 levels in the intracerebral hemorrhage progression. Bioengineered, 13(6), 14215–14226. https://doi.org/10.1080/21655979.2022.2084494
Coll, R. C., Schroder, K., & Pelegrín, P. (2022). Nlrp3 and pyroptosis blockers for treating inflammatory diseases. Trends in Pharmacological Sciences, 43(8), 653–668. https://doi.org/10.1016/j.tips.2022.04.003
Xiao, L., Dai, Z., Tang, W., Liu, C., Tang, B., Ajinkya, S., & Sase, A. (2021). Astragaloside iv alleviates cerebral ischemia-reperfusion injury through nlrp3 inflammasome-mediated pyroptosis inhibition via activating nrf2. Oxidative Medicine and Cellular Longevity, 2021, 9925514–9925561. https://doi.org/10.1155/2021/9925561
Wang, S., Yuan, Y., Chen, N., & Wang, H. (2019). The mechanisms of nlrp3 inflammasome/pyroptosis activation and their role in Parkinson’s disease. International Immunopharmacology, 67, 458–464. https://doi.org/10.1016/j.intimp.2018.12.019
Liu, B. H., Tu, Y., Ni, G. X., Yan, J., Yue, L., Li, Z. L., et al. (2021). Abelmoschus manihottotal flavones of ameliorates podocyte pyroptosis and injury in high glucose conditions by targeting mettl3-dependent ma modification-mediated nlrp3-inflammasome activation and pten/pi3k/akt signaling. Frontiers in Pharmacology, 12, 667644. https://doi.org/10.3389/fphar.2021.667644
Meng, L., Lin, H., Huang, X., Weng, J., Peng, F., & Wu, S. (2022). Mettl14 suppresses pyroptosis and diabetic cardiomyopathy by downregulating tincr lncrna. Cell Death & Disease, 13(1), 38. https://doi.org/10.1038/s41419-021-04484-z
Data availability statement
The datasets analyzed during the current study are available from the corresponding author on reasonable request.
Author information
Authors and Affiliations
Contributions
LL, JG, and WZ conceived the study; JG conducted the experiments; WZ analyzed the data; LL wrote the manuscript; all the authors read and approved the final version of the manuscript.
Corresponding author
Ethics declarations
Ethical Approval
No ethics approval was required for this study as it involved no human participants or animals.
Conflict of Interest
The authors declare no competing interests.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Li, L., Gong, J. & Zhang, W. Treatment of Intracerebral Hemorrhage with Traditional Chinese Medicine Monomer Wogonin by Modifying NLRP3 with METTL14 to Inhibit Neuronal Cell Pyroptosis. Appl Biochem Biotechnol (2024). https://doi.org/10.1007/s12010-023-04849-4
Accepted:
Published:
DOI: https://doi.org/10.1007/s12010-023-04849-4