Effects of nourishing qi, activating blood circulation, and inducing resuscitation on nerve cell pyroptosis after cerebral ischemia-reperfusion

Main Article Content

Lin-Quan Liu
Xiao-Ping Huang
Ya-Hong Cai
Yan She
Chang-Qing Deng

Keywords

BAP, cerebral ischemia-reperfusion, Nrf2/HO-1 signaling pathway, pyroptosis

Abstract

Cerebral ischemia-reperfusion (CIR) is a serious complication often associated with cerebral ischemia. The purpose of this study was to explore the therapeutic effect of nourishing qi, activating blood circulation, and inducing resuscitation (Borneol with astragaloside IV and Panax notoginseng total saponins, BAP) on CIR. Neurological function score system was used to determine the neurological function. The survival of nerve cells was detected by Nissl staining. The levels of IL-1β, IL-18, IL-4, and IL-10 were detected by ELISA. The expression of GSDMD, GSDMD-N, Nrf2, and HO-1 proteins in hippocampus tissues was measured by immunohistochemistry (IHC). Western blot, RT-qPCR, or immunofluorescence (IF) were used to detect the NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), pro-Caspase-1, Caspase-1, Nrf2, and HO-1 expression. Lactate dehydrogenase (LDH) level was analyzed by LDH release assay. Cell viability was determined by cell counting kit-8 (CCK8). Apoptosis was detected by flow cytometry. BAP significantly promoted the recovery of nerve function, the activity of nerve cells, and the expression of Nrf2, HO-1, IL-4, and IL-10 in rat hippocampus tissues after CIR. BAP has an obvious inhibitory effect on the expression of NLRP3, pro-Caspase-1, and Caspase-1 proteins, the release of IL-1β and IL-18 factors, and neuronal pyroptosis in hippocampal tissues. BAP also promoted IL-4 and IL-10 levels, and the activity of SH-SY5Y cells. The IL-1β, IL-18, NLRP3, pro-Caspase-1, Caspase-1, GSDMD, and GSDMD-N expressions were significantly inhibited by BAP in vitro, which was reversed by Nrf2 knockdown. This study confirmed that BAP alleviated rat CIR and inhibited the pyroptosis of SH-SY5Y cells by regulating the Nrf2/HO-1 signaling pathway. This study provided new directions and ideas for the treatment of CIR.

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References

Ahmed SMU, Luo L, Namani A, Wang XJ, Tang X. Nrf2 signaling pathway: Pivotal roles in inflammation. Biochim Biophys Acta Mol Basis Dis. 2017;1863(2):585−97. 10.1016/j.bbadis.2016.11.005

American Thoracic Society. Idiopathic pulmonary fibrosis: Diagnosis and treatment. International consensus statement. American Thoracic Society (ATS), and the European Respiratory Society (ERS). Am J Respir Crit Care Med. 2000;161(2 Pt 1):646−64. 10.1164/ajrccm.161.2.ats3-00

Broughton BRS, Reutens DC, Sobey CG. Apoptotic mechanisms after cerebral ischemia. Stroke. 2009;40(5):e331−9. 10.1161/STROKEAHA.108.531632

Cao X, Wang Y, Gao L. CHRFAM7A overexpression attenuates cerebral ischemia-reperfusion injury via inhibiting microglia pyroptosis mediated by the NLRP3/caspase-1 pathway. Inflammation. 2021;44(3):1023−34. 10.1007/s10753-020-01398-4

Chen J, Li Y, Wang L, Zhang Z, Lu D, Lu M, et al. Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke. 2001;32(4):1005−11. 10.1161/01.str.32.4.1005

Chen Z, Zhong H, Wei J, Lin S, Zong Z, Gong F, et al. Inhibition of Nrf2/HO-1 signaling leads to increased activation of the NLRP3 inflammasome in osteoarthritis. Arthritis Res Ther. 2019;21(1):300. 10.1186/s13075-019-2085-6

Dong Z, Pan K, Pan J, Peng Q, Wang Y. The possibility and molecular mechanisms of cell pyroptosis after cerebral ischemia. Neurosci Bull. 2018;34(6):1131−6. 10.1007/s12264-018-0294-7

Fan J, Liu D, He C, Li X, He F. Inhibiting adhesion events by Panax notoginseng saponins and Ginsenoside Rb1 protecting arteries via activation of Nrf2 and suppression of p38-VCAM-1 signal pathway. J Ethnopharmacol. 2016;192:423−30. 10.1016/j.jep.2016.09.022

Fan J, Lv H, Li J, Che Y, Xu B, Tao Z, et al. Roles of Nrf2/HO-1 and HIF-1α/VEGF in lung tissue injury and repair following cerebral ischemia/reperfusion injury. J Cell Physiol. 2019;234(6):7695−707. 10.1002/jcp.27767

Gao P, Du X, Liu L, Xu H, Liu M, Guan X, et al. Astragaloside IV alleviates tacrolimus-induced chronic nephrotoxicity via p62-Keap1-Nrf2 pathway. Front Pharmacol. 2020;11:610102. 10.3389/fphar.2020.610102

Gu D-M, Lu P-H, Zhang K, Wang X, Sun M, Chen G-Q, et al. EGFR mediates astragaloside IV-induced Nrf2 activation to protect cortical neurons against in vitro ischemia/reperfusion damages. Biochem Biophys Res Commun. 2015;457(3):391−7. 10.1016/j.bbrc.2015.01.002

Hou Y, Wang Y, He Q, Li L, Xie H, Zhao Y, et al. Nrf2 inhibits NLRP3 inflammasome activation through regulating Trx1/TXNIP complex in cerebral ischemia reperfusion injury. Behav Brain Res. 2018;336:32−9. 10.1016/j.bbr.2017.06.027

Huang Y, Yu J, Wan F, Zhang W, Yang H, Wang L, et al. Panaxatriol saponins attenuated oxygen-glucose deprivation injury in PC12 cells via activation of PI3K/Akt and Nrf2 signaling pathway. Oxid Med Cell Longev. 2014;2014:978034. 10.1155/2014/978034

Huang Z, Huang PL, Panahian N, Dalkara T, Fishman MC, Moskowitz MA. Effects of cerebral ischemia in mice deficient in neuronal nitric oxide synthase. Science. 1994;265(5180):1883−5. 10.1126/science.7522345

Iadecola C, Alexander M. Cerebral ischemia and inflammation. Curr Opin Neurol. 2001;14(1):89−94. 10.1097/00019052-200102000-00014

Ip FC-F, Zhao Y-M, Chan K-W, Cheng EY-L, Tong EP-S, Chandrashekar O, et al. Neuroprotective effect of a novel Chinese herbal decoction on cultured neurons and cerebral ischemic rats. BMC Complement Altern Med. 2016;16(1):437. 10.1186/s12906-016-1417-1

Jiang Q, Geng X, Warren J, Eugene Paul Cosky E, Kaura S, Stone C, et al. Hypoxia inducible factor-1α (HIF-1α) mediates NLRP3 inflammasome-dependent-pyroptotic and apoptotic cell death following ischemic stroke. Neuroscience. 2020;448:126−39. 10.1016/j.neuroscience.2020.09.036

Khandelwal P, Yavagal DR, Sacco RL. Acute ischemic stroke intervention. J Am Coll Cardiol. 2016;67(22):2631−44. 10.1016/j.jacc.2016.03.555

Li L, Hou X, Xu R, Liu C, Tu M. Research review on the pharmacological effects of astragaloside IV. Fundam Clin Pharmacol. 2017;31(1):17−36. 10.1111/fcp.12232

Li L, Shu M-Q, Chen J. CYLD deficiency exacerbates lipopolysaccharide (LPS)-induced pyroptosis in astrocytes of mice with sepsis. Biochem Biophys Res Commun. 2019;514(4):1066−73. 10.1016/j.bbrc.2019.05.033

Li X, An P, Han F, Yu M, Yu Z, Li Y. Silencing of YTHDF1 attenuates cerebral stroke by inducing PTEN degradation and activating the PTEN/AKT/mTOR pathway. Mol Biotechnol. 2022;??:??. 10.1007/s12033-022-00575-0

Li Y, Ren M, Wang J, Ma R, Chen H, Xie Q, et al. Progress in borneol intervention for ischemic stroke: A systematic review. Front Pharmacol. 2021;12:606682. 10.3389/fphar.2021.606682

Li YQ, Jin M, Qiu SL, Wang PL, Zhu TG, Wang CL, et al. Effect of Chinese drugs for supplementing Qi, nourishing Yin and activating blood circulation on myocardial perfusion in patients with acute myocardial infarction after revascularization. Chin J Integr Med. 2009;15(1):19−25. 10.1007/s11655-009-0019-7

Liu R, Zhang L, Lan X, Li L, Zhang TT, Sun JH, et al. Protection by borneol on cortical neurons against oxygen-glucose deprivation/reperfusion: Involvement of anti-oxidation and anti-inflammation through nuclear transcription factor κappaB signaling pathway. Neuroscience. 2011;176:408−19. 10.1016/j.neuroscience.2010.11.029

Liu HY, Wang W, Shi DZ, Ge JB, Zhang L, Peng J, et al. Protective effect of Chinese herbs for supplementing qi, nourishing yin and activating blood circulation on heart function of patients with acute coronary syndrome after percutaneous coronary intervention. Chin J Integr Med. 2012;18(6):423−30. 10.1007/s11655-012-0966-5

Liu SH, Zhang YC. Effect of levocarnitine on cerebral ischemia-reperfusion rats via activating Nrf2/ARE signaling pathway. Eur Rev Med Pharmacol Sci. 2019;23(18):8168−74. 10.26355/eurrev_201909_19037

Lu Y, Chen X, Du S, Wu Q, Yao Z, Zhai Y. The in situ and in vivo study on enhancing effect of borneol in nasal absorption of Geniposide in rats. Arch Pharm Res. 2010;33(5):691−6. 10.1007/s12272-010-0507-8

Pang Y, Zhang P-C, Lu R-R, Li H-L, Li J-C, Fu H-X, et al. Andrade-oliveira salvianolic acid B modulates caspase-1-mediated pyroptosis in renal ischemia-reperfusion injury Nrf2 pathway. Front Pharmacol. 2020;11:541426. 10.3389/fphar.2020.541426

Ren J, Zou M, Gao P, Wang Y, Cheng G. Tissue distribution of borneol-modified ganciclovir-loaded solid lipid nanoparticles in mice after intravenous administration. Eur J Pharm Biopharm. 2013;83(2):141−8. 10.1016/j.ejpb.2012.10.018

Shi M, Chen J, Liu T, Dai W, Zhou Z, Chen L, et al. Protective effects of remimazolam on cerebral ischemia/reperfusion injury in rats by inhibiting of NLRP3 inflammasome-dependent pyroptosis. Drug Des Devel Ther. 2022;16:413−23. 10.2147/dddt.S344240

Shi X, Yu W, Yang T, Liu W, Zhao Y, Sun Y, et al. Panax notoginseng saponins provide neuroprotection by regulating NgR1/RhoA/ROCK2 pathway expression, in vitro and in vivo. J Ethnopharmacol. 2016;190:301−12. 10.1016/j.jep.2016.06.017

Sorrells SF, Paredes MF, Cebrian-Silla A, Sandoval K, Qi D, Kelley KW, et al. Human hippocampal neurogenesis drops sharply in children to undetectable levels in adults. Nature. 2018;555(7696):377−81. 10.1038/nature25975

Tang B, She X, Deng C-Q. Effect of the combination of astragaloside IV and saponins on pyroptosis and necroptosis in rat models of cerebral ischemia-reperfusion. Exp Ther Med. 2021;22(4):1123. 10.3892/etm.2021.10557

Tang H, Gamdzyk M, Huang L, Gao L, Lenahan C, Kang R, et al. Delayed recanalization after MCAO ameliorates ischemic stroke by inhibiting apoptosis via HGF/c-Met/STAT3/Bcl-2 pathway in rats. Exp Neurol. 2020;330:113359. 10.1016/j.expneurol.2020.113359

Turovsky EA, Golovicheva VV, Varlamova EG, Danilina TI, Goryunov KV, Shevtsova YA, et al. Mesenchymal stromal cell-derived extracellular vesicles afford neuroprotection by modulating PI3K/AKT pathway and calcium oscillations. Int J Biol Sci. 2022;18(14):5345−68. 10.7150/ijbs.73747

Wardyn JD, Ponsford AH, Sanderson CM. Dissecting molecular cross-talk between Nrf2 and NF-κB response pathways. Biochem Soc Trans. 2015;43(4):621−6. 10.1042/BST20150014

Wu L-M, Wu S-G, Chen F, Wu Q, Wu C-M, Kang C-M, et al. Atorvastatin inhibits pyroptosis through the lncRNA NEXN-AS1/NEXN pathway in human vascular endothelial cells. Atherosclerosis. 2020;293:26−34. 10.1016/j.atherosclerosis.2019.11.033

Xing C, Yan G, Liu Q. Inhibition of GPR4 attenuates SH-SY5Y cell injury in cerebral ischemia/reperfusion via anti-apoptotic pathways. Acta Biochim Pol. 2021;68(2):181–6. 10.18388/abp.2020_5497

Xu D, Huang P, Yu Z, Xing DH, Ouyang S, Xing G. Efficacy and safety of panax notoginseng saponin therapy for acute intracerebral hemorrhage, meta-analysis, and mini review of potential mechanisms of action. Front Neurol. 2014;5:274. 10.3389/fneur.2014.00274

Xu X, Zhang L, Ye X, Hao Q, Zhang T, Cui G, et al. Nrf2/ARE pathway inhibits ROS-induced NLRP3 inflammasome activation in BV2 cells after cerebral ischemia reperfusion. Inflamm Res. 2018;67(1):57−65. 10.1007/s00011-017-1095-6

Ya B-L, Liu Q, Li H-F, Cheng H-J, Yu T, Chen L, et al. Uric acid protects against focal cerebral ischemia/reperfusion-induced oxidative stress via activating Nrf2 and regulating neurotrophic factor expression. Oxid Med Cell Longev. 2018;2018:6069150. 10.1155/2018/6069150

Yan Z, Qi W, Zhan J, Lin Z, Lin J, Xue X, et al. Activating Nrf2 signalling alleviates osteoarthritis development by inhibiting inflammasome activation. J Cell Mol Med. 2020;24(22):13046−57. 10.1111/jcmm.15905

Yang C, Mo Y, Xu E, Wen H, Wei R, Li S, et al. Astragaloside IV ameliorates motor deficits and dopaminergic neuron degeneration via inhibiting neuroinflammation and oxidative stress in a Parkinson’s disease mouse model. Int Immunopharmacol. 2019;75:105651. 10.1016/j.intimp.2019.05.036

Yang J, Gao F, Zhang Y, Liu Y, Zhang D. Buyang Huanwu Decoction (BYHWD) enhances angiogenic effect of mesenchymal stem cell by upregulating vegf expression after focal cerebral ischemia. J Mol Neurosci. 2015;56(4):898−906. 10.1007/s12031-015-0539-0

Yang X, Xiong X, Wang H, Wang J. Protective effects of panax notoginseng saponins on cardiovascular diseases: A comprehensive overview of experimental studies. Evid Based Complement Alternat Med. 2014;2014:204840. 10.1155/2014/204840

Yu B, Ruan M, Cui X-b, Guo J-M, Xu L, Dong X-P. Effects of borneol on the pharmacokinetics of geniposide in cortex, hippocampus, hypothalamus and striatum of conscious rat by simultaneous brain microdialysis coupled with UPLC-MS. J Pharm Biomed Anal. 2013;77:128−32. 10.1016/j.jpba.2013.01.017

Zhang D, Qian J, Zhang P, Li H, Shen H, Li X, et al. Gasdermin D serves as a key executioner of pyroptosis in experimental cerebral ischemia and reperfusion model both in vivo and in vitro. J Neurosci Res. 2019;97(6):645−60. 10.1002/jnr.24385

Zhang J, Wu C, Gao L, Du G, Qin X. Astragaloside IV derived from Astragalus membranaceus: A research review on the pharmacological effects. Adv Pharmacol. 2020;87:89−112. 10.1016/bs.apha.2019.08.002

Zhang L, Liu H, Jia L, Lyu J, Sun Y, Yu H, et al. Exosomes mediate hippocampal and cortical neuronal injury induced by hepatic ischemia-reperfusion injury through activating pyroptosis in rats. Oxid Med Cell Longev. 2019;2019:3753485. 10.1155/2019/3753485

Zhang R, Liu Y, Zhang C, Ma M, Li S, Hong Y. Salt-inducible kinase 2 regulates energy metabolism in rats with cerebral ischemia-reperfusion. Zhejiang Da Xue Xue Bao Yi Xue Ban. 2021;50(3):352−60. 10.3724/zdxbyxb-2021-0164

Zhang W-W, Xu F, Wang D, Ye J, Cai S-Q. Buyang Huanwu Decoction ameliorates ischemic stroke by modulating multiple targets with multiple components: In vitro evidences. Chin J Nat Med. 2018;16(3):194−202. 10.1016/S1875-5364(18)30047-5

Zhang X, Wu J, Zhang B. Xuesaitong injection as one adjuvant treatment of acute cerebral infarction: A systematic review and meta-analysis. BMC Complement Altern Med. 2015;15:36. 10.1186/s12906-015-0560-4

Zhao M-W, Yang P, Zhao L-L. Chlorpyrifos activates cell pyroptosis and increases susceptibility on oxidative stress-induced toxicity by miR-181/SIRT1/PGC-1α/Nrf2 signaling pathway in human neuroblastoma SH-SY5Y cells: Implication for association between chlorpyrifos and Parkinson’s disease. Environ Toxicol. 2019;34(6):699−707. 10.1002/tox.22736

Zheng W, Matei N, Pang J, Luo X, Song Z, Tang J, et al. Delayed recanalization at 3 days after permanent MCAO attenuates neuronal apoptosis through FGF21/FGFR1/PI3K/Caspase-3 pathway in rats. Exp Neurol. 2019;320:113007. 10.1016/j.expneurol.2019.113007

Zheng X-W, Shan C-S, Xu Q-Q, Wang Y, Shi Y-H, Wang Y, et al. Buyang Huanwu Decoction Ttrgets SIRT1/VEGF pathway to promote angiogenesis after cerebral ischemia/reperfusion injury. Front Neurosci. 2018;12:911. 10.3389/fnins.2018.00911

Zhu Q-Y, Tang S, Yang X-Q, Ding H, Liu X-D, Zeng X-B, et al. Borneol enhances the protective effect against cerebral ischemia/reperfusion injury by promoting the access of astragaloside IV and the components of Panax notoginseng saponins into the brain. Phytomedicine. 2022;94:153822. 10.1016/j.phymed.2021.153822