Protective effects of andrographolide against cerebral ischemia‑reperfusion injury in mice

Li,Yan; Xiang,Li-Ling; Miao,Jin-Xin; Miao,Ming-San; Wang,Can

doi:10.3892/ijmm.2021.5019

Protective effects of andrographolide against cerebral ischemia‑reperfusion injury in mice

Authors:
- Yan Li
- Li-Ling Xiang
- Jin-Xin Miao
- Ming-San Miao
- Can Wang
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Affiliations: College of Traditional Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou, Henan 450046, P.R. China, School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, Henan 450046, P.R. China
Published online on: August 3, 2021 https://doi.org/10.3892/ijmm.2021.5019
Article Number: 186
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ischemic stroke is one of the most common causes of mortality worldwide and is a primary cause of disability and mortality in adults. There is an unmet need for drugs that can effectively treat ischemic stroke. Hence, the present study explored the neuroprotective effects of andrographolide (Andro) in a mouse model of bilateral common carotid artery occlusion, and systematically evaluated the potential mechanisms underlying its effects. The effects of Andro on mouse brain tissue following cerebral ischemia‑reperfusion injury (CIRI) were evaluated by histopathological (H&E and Nissl) and immunofluorescence [glial fibrillary acidic protein (GFAP) and neuronal nuclei (NeuN)] staining. A traditional Chinese medicine‑based network pharmacology method was performed to establish and analyze compound‑target‑disease and function‑pathway networks in order to elucidate the possible mechanisms responsible for the protective role of Andrographis paniculata in CIRI. In addition, western blot analysis and RT‑qPCR was performed to evaluate the expression and activation of signaling proteins predicted to be involved in this mechanism. The amelioration of histopathological alterations was observed in mice pre‑treated with Andro. Immunofluorescence staining revealed that Andro decreased the expression of GFAP and increased the expression of NeuN, and significantly decreased the levels of pro‑inflammatory cytokines (IL‑1β, IL‑6 and TNF‑α). Network pharmacology analysis revealed that neuroinflammatory response and apoptosis were associated with the effects of Andrographis paniculata on CIRI. Western blot analysis revealed that the mice pre‑treated with Andro exhibited an upregulated protein expression of tropomyosin receptor kinase B (TrkB), p‑PI3K and p‑Akt, as well as a decrease in the expression of GFAP and an increase in the expression of NeuN. In addition, the data of RT‑qPCR indicated that the mice pre‑treated with Andro exhibited a significantly decreased expression of encoding IL‑1β mRNA, IL‑6 mRNA and TNF‑α mRNA in the brain compared to the untreated mice following CIRI. On the whole, the findings of the present study suggest that pre‑treatment with Andro exerts a protective effect against CIRI, which may be partly related to its potential to reduce neuroinflammatory response and apoptosis in patients with stroke.

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1	Krishnamurthi RV, Feigin VL, Forouzanfar MH, Mensah GA, Connor M, Bennett DA, Moran AE, Sacco RL, Anderson LM, Truelsen T, et al: Global and regional burden of first-ever ischaemic and haemorrhagic stroke during 1990-2010: Findings from the global burden of disease study 2010. Lancet Glob Health. 1:e259–e281. 2013. View Article : Google Scholar
2	Miralbell J, López-Cancio E, López-Oloriz J, Arenillas JF, Barrios M, Soriano-Raya JJ, Galán A, Cáceres C, Alzamora M, Pera G, et al: Cognitive patterns in relation to biomarkers of cerebrovascular disease and vascular risk factors. Cerebrovasc Dis. 36:98–105. 2013. View Article : Google Scholar : PubMed/NCBI
3	Doyle KP, Simon RP and Stenzel-Poore MP: Mechanisms of ischemic brain damage. Neuropharmacology. 55:310–318. 2008. View Article : Google Scholar : PubMed/NCBI
4	Song W, Zhao J, Yan XS, Fang X, Huo DS, Wang H, Jia JX and Yang ZJ: Mechanisms associated with protective effects of ginkgo biloba leaf extracton in rat cerebral ischemia reperfusion injury. J Toxicol Environ Health A. 82:1045–1051. 2019. View Article : Google Scholar : PubMed/NCBI
5	Becker KJ and Buckwalter M: Stroke, inflammation and the immune response: Dawn of a new era. Neurotherapeutics. 13:659–660. 2016. View Article : Google Scholar : PubMed/NCBI
6	Liu T, Yang K, Li G, Zhou K, Tan J, Chen J, Li T, Yu Y and Ning W: Experimental evidence and network pharmacology identify the molecular targets of Tong Sheng tablets in cerebral ischemia reperfusion injury. Am J Transl Res. 11:3301–3316. 2019.PubMed/NCBI
7	Iadecola C and Anrather J: The immunology of stroke: From mechanisms to translation. Nat Med. 17:796–808. 2011. View Article : Google Scholar : PubMed/NCBI
8	Benjamin EJ, Blaha MJ, Chiuve SE, Cushman M, Das SR, Deo R, de Ferranti SD, Floyd J, Fornage M, Gillespie C, et al: Heart disease and stroke statistics-2017 update: A report from the American heart association. Circulation. 135:e146–e603. 2017. View Article : Google Scholar
9	Powers WJ and Rabinstein AA: Response by powers and rabinstein to letter regarding article, '2018 guidelines for the early management of patients with acute ischemic stroke: A guideline for healthcare professionals from the American heart association/American stroke association'. Stroke. 50:e277–e278. 2019. View Article : Google Scholar
10	Muresanu DF, Strilciuc S and Stan A: Current drug treatment of acute ischemic stroke: Challenges and opportunities. CNS Drugs. 33:841–847. 2019. View Article : Google Scholar : PubMed/NCBI
11	Chan SJ, Wong WS, Wong PT and Bian JS: Neuroprotective effects of andrographolide in a rat model of permanent cerebral ischaemia. Br J Pharmacol. 161:668–679. 2010. View Article : Google Scholar : PubMed/NCBI
12	Wang DP, Yin H, Lin Q, Fang SP, Shen JH, Wu YF, Su SH and Hai J: Andrographolide enhances hippocampal BDNF signaling and suppresses neuronal apoptosis, astroglial activation, neuroinflammation, and spatial memory deficits in a rat model of chronic cerebral hypoperfusion. Naunyn Schmiedebergs Arch Pharmacol. 392:1277–1284. 2019. View Article : Google Scholar : PubMed/NCBI
13	Yuan H, Ma Q, Cui H, Liu G, Zhao X, Li W and Piao G: How can synergism of traditional medicines benefit from network pharmacology? Molecules. 22:11352017. View Article : Google Scholar
14	Ning K, Zhao X, Poetsch A, Chen WH and Yang J: Computational molecular networks and network pharmacology. Biomed Res Int. 2017:75739042017. View Article : Google Scholar
15	Zhang JJ, Gao TT, Wang Y, Wang JL, Guan W, Wang YJ, Wang CN, Liu JF and Jiang B: Andrographolide exerts significant antidepressant-like effects involving the hippocampal BDNF system in mice. Int J Neuropsychopharmacol. 22:585–600. 2019. View Article : Google Scholar
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
17	Fan M, Song C, Wang T, Li L, Dong Y, Jin W and Lu P: Protective effects of lithium chloride treatment on repeated cerebral ischemia-reperfusion injury in mice. Neurol Sci. 36:315–321. 2015. View Article : Google Scholar
18	Huang DW, Sherman BT, Tan Q, Kir J, Liu D, Bryant D, Guo Y, Stephens R, Baseler MW, Lane HC and Lempicki RA: DAVID bioinformatics resources: Expanded annotation database and novel algorithms to better extract biology from large gene lists. Nucleic Acids Res. 35(Web Server Issue): W169–W175. 2007. View Article : Google Scholar : PubMed/NCBI
19	Schooling CM, Huang JV, Zhao JV, Kwok MK, Au Yeung SL and Lin SL: Disconnect between genes associated with ischemic heart disease and targets of ischemic heart disease treatments. EBioMedicine. 28:311–315. 2018. View Article : Google Scholar : PubMed/NCBI
20	van Hoof RHM, Schreuder FHBM, Nelemans P, Truijman MTB, van Orshoven NP, Schreuder TH, Mess WH, Heeneman S, van Oostenbrugge RJ, Wildberger JE and Kooi ME: Ischemic stroke patients demonstrate increased carotid plaque microvasculature compared to (Ocular) transient ischemic attack patients. Cerebrovasc Dis. 44:297–303. 2017. View Article : Google Scholar
21	Zhang Q, An R, Tian X, Yang M, Li M, Lou J, Xu L and Dong Z: β-Caryophyllene pretreatment alleviates focal cerebral ischemia-reperfusion injury by activating PI3K/Akt signaling pathway. Neurochem Res. 42:1459–1469. 2017. View Article : Google Scholar : PubMed/NCBI
22	Zhang X, Du Q, Yang Y, Wang J, Liu Y, Zhao Z, Zhu Y and Liu C: Salidroside alleviates ischemic brain injury in mice with ischemic stroke through regulating BDNK mediated PI3K/Akt pathway. Biochem Pharmacol. 156:99–108. 2018. View Article : Google Scholar
23	Yen TL, Chen RJ, Jayakumar T, Lu WJ, Hsieh CY, Hsu MJ, Yang CH, Chang CC, Lin YK, Lin KH and Sheu JR: Andrographolide stimulates p38 mitogen-activated protein kinase-nuclear factor erythroid-2-related factor 2-heme oxygenase 1 signaling in primary cerebral endothelial cells for definite protection against ischemic stroke in rats. Transl Res. 170:57–72. 2016. View Article : Google Scholar
24	Chern CM, Liou KT, Wang YH, Liao JF, Yen JC and Shen YC: Andrographolide inhibits PI3K/AKT-dependent NOX2 and iNOS expression protecting mice against hypoxia/ischemia-induced oxidative brain injury. Planta Med. 77:1669–1679. 2011. View Article : Google Scholar : PubMed/NCBI
25	Wada T and Penninger JM: Mitogen-activated protein kinases in apoptosis regulation. Oncogene. 23:2838–2849. 2004. View Article : Google Scholar
26	Yu ZH, Cai M, Xiang J, Zhang ZN, Zhang JS, Song XL, Zhang W, Bao J, Li WW and Cai DF: PI3K/Akt pathway contributes to neuroprotective effect of Tongxinluo against focal cerebral ischemia and reperfusion injury in rats. J Ethnopharmacol. 181:8–19. 2016. View Article : Google Scholar : PubMed/NCBI
27	Bao Y, Qin L, Kim E, Bhosle S, Guo H, Febbraio M, Haskew-Layton RE, Ratan R and Cho S: CD36 is involved in astrocyte activation and astroglial scar formation. J Cereb Blood Flow Metab. 32:1567–1577. 2012. View Article : Google Scholar : PubMed/NCBI
28	Freeman MR: Specification and morphogenesis of astrocytes. Science. 330:774–778. 2010. View Article : Google Scholar : PubMed/NCBI
29	Lanfranconi S, Locatelli F, Corti S, Candelise L, Comi GP, Baron PL, Strazzer S, Bresolin N and Bersano A: Growth factors in ischemic stroke. J Cell Mol Med. 15:1645–1687. 2011. View Article : Google Scholar
30	Erpolat S, Celik HT and Bozkurt B: Brain-derived neurotrophic factor is increased in serum levels of patients with symptomatic dermographism. Postepy Dermatol Alergol. 34:346–349. 2017. View Article : Google Scholar : PubMed/NCBI
31	Ya BL, Liu Q, Li HF, Cheng HJ, Yu T, Chen L, Wang Y, Yuan LL, Li WJ, Liu WY and Bai B: Uric acid protects against focal cerebral ischemia/reperfusion-induced oxidative stress via activating Nrf2 and regulating neurotrophic factor expression. Oxid Med Cell Longev. 2018:60691502018. View Article : Google Scholar :
32	Wang J, Ma W and Liu Y: Long non-coding RNA HULC promotes bladder cancer cells proliferation but inhibits apoptosis via regulation of ZIC2 and PI3K/AKT signaling pathway. Cancer Biomark. 20:425–434. 2017. View Article : Google Scholar : PubMed/NCBI
33	Franke TF, Kaplan DR and Cantley LC: PI3K: Downstream AKTion blocks apoptosis. Cell. 88:435–437. 1997. View Article : Google Scholar
34	Calleja V, Alcor D, Laguerre M, Park J, Vojnovic B, Hemmings BA, Downward J, Parker PJ and Larijani B: Intramolecular and intermolecular interactions of protein kinase B define its activation in vivo. PLoS Biol. 5:e952007. View Article : Google Scholar