Astragaloside IV attenuates cerebral ischemia‑reperfusion injury in rats through the inhibition of calcium‑sensing receptor‑mediated apoptosis

Du,Shu‑Jin; Zhang,Ying; Zhao,Yan‑Meng; Dong,Ya‑Jie; Tang,Jing‑Long; Zhou,Xiao‑Hong; Gao,Wei‑Juan

doi:10.3892/ijmm.2020.4777

Astragaloside IV attenuates cerebral ischemia‑reperfusion injury in rats through the inhibition of calcium‑sensing receptor‑mediated apoptosis

Authors:
- Shu‑Jin Du
- Ying Zhang
- Yan‑Meng Zhao
- Ya‑Jie Dong
- Jing‑Long Tang
- Xiao‑Hong Zhou
- Wei‑Juan Gao
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Affiliations: Hebei Key Laboratory of Chinese Medicine Research on Cardio‑Cerebrovascular Disease, Hebei University of Chinese Medicine, Shijiazhuang, Hebei 050200, P.R. China, Department of Pathophysiology, Chengde Medical College, Chengde, Hebei 067000, P.R. China, The Reproductive Genetics Laboratory, Jinan Adicon Clinical Labor, Binzhou, Shandong 256600, P.R. China
Published online on: November 2, 2020 https://doi.org/10.3892/ijmm.2020.4777
Pages: 302-314
Copyright: © Du et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cerebral ischemia‑reperfusion injury (CIRI), caused by the reperfusion of blocked vessels following ischemic stroke, can lead to secondary brain injury. Throughout CIRI, apoptosis serves an important role. Astragaloside IV is a potential neuroprotectant that alleviates CIRI by inhibiting apoptosis. The calcium‑sensing receptor (CaSR) is a G‑protein‑coupled receptor, the activation of which aggravates ischemia‑reperfusion injury. The aim of the present study was to investigate whether the protective effect of Astragaloside IV on CIRI may be associated with the regulation of CaSR. A rat middle cerebral artery occlusion/reperfusion (MCAO/R) model and an oxygen and glucose deprivation/reoxygenation (OGD/R) model of pheochromocytoma (PC12) cells were used to study the neuronal injury induced by CIRI. Neurological function scores (NFS), 2,3,5‑triphe‑nylterazolium chloride and hematoxylin and eosin staining were used to determine brain damage in rats. Cell viability was measured to evaluate the injury of OGD/R PC12 cells. Western blotting was used to examine the expression of proteins associated with apoptosis and CaSR. The CaSR antagonist NPS‑2143 and agonist GdCl3 were used to further confirm the effects of CaSR during the process of apoptosis. The results demonstrated that Astragaloside IV alleviated CIRI by decreasing the NFS of rats, reducing the infarction volume of the brain and promoting the viability of PC12 cells, as well as inhibiting the expression of cleaved caspase‑3 and CaSR, which was induced by CIRI. The results of the present study suggested that the activation of CaSR may be involved in CIRI‑induced brain damage in rats, and that Astragaloside IV may alleviate CIRI by inhibiting CaSR activation‑induced apoptosis.

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1	Hankey GJ: Stroke. Lancet. 389:641–654. 2017. View Article : Google Scholar
2	Writing Group Members; Mozaffarian D, Benjamin EJ, Go AS, Arnett DK, Blaha MJ, Cushman M, Das SR, de Ferranti S, Després JP, et al: Heart disease and stroke statistics-2016 update: A report from the American heart association. Circulation. 133:e38–e360. 2016.
3	Powers WJ, Rabinstein AA, Ackerson T, Adeoye OM, Bambakidis NC, Becker K, Biller J, Brown M, Demaerschalk BM, Hoh B, et al: Guidelines for the early management of patients with acute ischemic stroke: 2019 update to the 2018 guidelines for the early management of acute ischemic stroke: A guideline for healthcare professionals from the American heart association/American stroke association. Stroke. 50:e344–e418. 2019. View Article : Google Scholar
4	Qureshi AI, Singh B, Huang W, Du Z, Lobanova I, Liaqat J and Siddiq F: Mechanical thrombectomy in acute ischemic stroke patients performed within and outside clinical trials in the United States. Neurosurgery. 86:E2–E8. 2020.
5	Chen YW, Sung SF, Chen CH, Tang SC, Tsai LK, Lin HJ, Huang HY, Po HL, Sun Y, Chen PL, et al: Intravenous thrombolysis administration 3-4.5 h after acute ischemic stroke: A retrospective, multicenter study. Front Neurol. 10:10382019. View Article : Google Scholar : PubMed/NCBI
6	Chen CH and Hsieh CL: Effect of acupuncture on oxidative stress iduced by cerebral ischemia-reperfusion injury. Antioxidants (Basel). 9:2482020. View Article : Google Scholar
7	Wu L, Xiong X, Wu X, Ye Y, Jian Z, Zhi Z and Gu L: Targeting oxidative stress and inflammation to prevent ischemia-reperfusion injury. Front Mol Neurosci. 13:282020. View Article : Google Scholar : PubMed/NCBI
8	Khandelwal P, Yavagal DR and Sacco RL: Acute ischemic stroke intervention. J Am Coll Cardiol. 67:2631–2644. 2016. View Article : Google Scholar : PubMed/NCBI
9	Zhang Y, Zhang Y, Jin XF, Zhou XH, Dong XH, Yu WT and Gao WJ: The role of astragaloside IV against cerebral ischemia/reperfusion injury: Suppression of apoptosis via promotion of P62-LC3-autophagy. Molecules. 24:18382019. View Article : Google Scholar :
10	Liu D, Gu Y, Wang W and Chen W: Astragalin alleviates ischemia/reperfusion-induced brain injury via suppression of endoplasmic reticulum stress. Mol Med Rep. 22:4070–4078. 2020.PubMed/NCBI
11	Liu Y, Yang H, Jia G, Li L, Chen H, Bi J and Wang C: The synergistic neuroprotective effects of combined rosuvastatin and resveratrol pretreatment against cerebral ischemia/reperfusion injury. J Stroke Cerebrovasc Dis. 27:1697–1704. 2018. View Article : Google Scholar : PubMed/NCBI
12	Gu JH, Ge JB, Li M, Wu F, Zhang W and Qin ZH: Inhibition of NF-κB activation is associated with anti-inflammatory and anti-apoptotic effects of ginkgolide B in a mouse model of cerebral ischemia/reperfusion injury. Eur J Pharm Sci. 47:652–660. 2012. View Article : Google Scholar : PubMed/NCBI
13	Zhou H, Yang WS, Li Y, Ren T, Peng L, Guo H, Liu JF, Zhou Y, Zhao Y, Yang LC and Jin X: Oleoylethanolamide attenuates apoptosis by inhibiting the TLR4/NF-κB and ERK1/2 signaling pathways in mice with acute ischemic stroke. Naunyn Schmiedebergs Arch Pharmacol. 390:77–84. 2017. View Article : Google Scholar
14	Naito MG, Xu D, Amin P, Lee J, Wang H, Li W, Kelliher M, Pasparakis M and Yuan J: Sequential activation of necroptosis and apoptosis cooperates to mediate vascular and neural pathology in stroke. Proc Natl Acad Sci USA. 117:4959–4970. 2020. View Article : Google Scholar : PubMed/NCBI
15	Bai X, Tan TY, Li YX, Li Y, Chen YF, Ma R, Wang SY, Li Q and Liu ZQ: The protective effect of cordyceps sinensis extract on cerebral ischemic injury via modulating the mitochondrial respiratory chain and inhibiting the mitochondrial apoptotic pathway. Biomed Pharmacother. 124:1098342020. View Article : Google Scholar : PubMed/NCBI
16	Wang HL, Zhou QH, Xu MB, Zhou XL and Zheng GQ: Astragaloside IV for experimental focal cerebral ischemia: Preclinical evidence and possible mechanisms. Oxid Med Cell Longev. 2017:84243262017. View Article : Google Scholar : PubMed/NCBI
17	Lu FH, Tian Z, Zhang WH, Zhao YJ, Li HL, Ren H, Zheng HS, Liu C, Hu GX, Tian Y, et al: Calcium-sensing receptors regulate cardiomyocyte Ca²⁺ signaling via the sarcoplasmic reticulum-mitochondrion interface during hypoxia/reoxygenation. J Biomed Sci. 17:502010. View Article : Google Scholar
18	Yin F, Zhou H, Fang Y, Li C, He Y, Yu L, Wan H and Yang J: Astragaloside IV alleviates ischemia reperfusion-induced apoptosis by inhibiting the activation of key factors in death receptor pathway and mitochondrial pathway. J Ethnopharmacol. 248:1123192020. View Article : Google Scholar
19	Yin B, Hou XW and Lu ML: Astragaloside IV attenuates myocardial ischemia/reperfusion injury in rats via inhibition of calcium-sensing receptor-mediated apoptotic signaling path-ways. Acta Pharmacol Sin. 40:599–607. 2019. View Article : Google Scholar
20	Zhang Y, Qiao L, Xu W, Wang X, Li H, Xu W, Chu K and Lin Y: Paeoniflorin attenuates cerebral ischemia-induced injury by regulating Ca²⁺/CaMKII/CREB signaling pathway. Molecules. 22:3592017. View Article : Google Scholar
21	Yan L, Zhu T, Sun T, Wang L, Pan S, Tao Z, Yang Z and Cao K: Activation of calcium-sensing receptors is associated with apoptosis in a model of simulated cardiomyocytes ischemia/reperfusion. J Biomed Res. 24:301–307. 2010. View Article : Google Scholar : PubMed/NCBI
22	Kienitz MC, Niemeyer A, Konig GM, Kostenis E, Pott L and Rinne A: Biased signaling of Ca²⁺-sensing receptors in cardiac myocytes regulates GIRK channel activity. J Mol Cell Cardiol. 130:107–121. 2019. View Article : Google Scholar : PubMed/NCBI
23	Zhen Y, Ding C, Sun J, Wang Y, Li S and Dong L: Activation of the calcium-sensing receptor promotes apoptosis by modulating the JNK/p38 MAPK pathway in focal cerebral ischemia-reperfusion in mice. Am J Transl Res. 8:911–921. 2016.PubMed/NCBI
24	Zhang ZL, Li ZR, Li JS and Wang SR: Calcium-sensing receptor antagonist NPS-2143 suppresses proliferation and invasion of gastric cancer cells. Cancer Gene Ther. 27:548–557. 2019. View Article : Google Scholar : PubMed/NCBI
25	Pearce SH and Thakker RV: The calcium sensing receptor: Insights into extracellular calcium homeostasis in health and disease. J Endocrinol. 154:371–378. 1997. View Article : Google Scholar : PubMed/NCBI
26	Chattopadhyay N, Vassilev PM and Brown EW: Calcium-sensing receptor: Roles in and beyond systemic calcium homeostasis. Biol Chem. 378:759–768. 1997.PubMed/NCBI
27	Meng D, Chen XJ, Bian YY, Li P, Yang D and Zhang JN: Effect of astragalosides on intracellular calcium overload in cultured cardiac myocytes of neonatal rats. Am J Chin Med. 33:11–20. 2005. View Article : Google Scholar : PubMed/NCBI
28	Hu B, Tong F, Xu L, Shen Z, Yan L, Xu G and Shen R: Role of calcium sensing receptor in streptozotocin-induced diabetic rats exposed to renal ischemia reperfusion injury. Kidney Blood Press Res. 43:276–286. 2018. View Article : Google Scholar : PubMed/NCBI
29	Pan R, Cai J, Zhan L, Guo Y, Huang RY, Li X, Zhou M, Xu D, Zhan J and Chen H: Buyang Huanwu decoction facilitates neuro-rehabilitation through an improvement of synaptic plasticity in cerebral ischemic rats. BMC Complement Altern Med. 17:1732017. View Article : Google Scholar
30	Zheng XW, Shan CS, Xu QQ, Wang Y, Shi YH, Wang Y and Zheng GQ: Buyang Huanwu decoction targets SIRT1/VEGF pathway to promote angiogenesis after cerebral ischemia/reperfusion injury. Front Neurosci. 12:9112018. View Article : Google Scholar : PubMed/NCBI
31	Qi LW, Yu QT, Yi L, Ren MT, Wen XD, Wang YX and Li P: Simultaneous determination of 15 marker constituents in various radix Astragali preparations by solid-phase extraction and high-performance liquid chromatography. J Sep Sci. 31:97–106. 2008. View Article : Google Scholar
32	Sun WX, Zhang ZF, Xie J, He Y, Cheng Y, Ding LS, Luo P and Qing LS: Determination of an astragaloside IV derivative LS-102 in plasma by ultra-performance liquid chromatography-tandem mass spectrometry in dog plasma and its application in a pharmacokinetic study. Phytomedicine. 53:243–251. 2019. View Article : Google Scholar : PubMed/NCBI
33	Qu YZ, Li M, Zhao YL, Zhao ZW, Wei XY, Liu JP, Gao L and Gao GD: Astragaloside IV attenuates cerebral ischemia-reperfusion-induced increase in permeability of the blood-brain barrier in rats. Eur J Pharmacol. 606:137–141. 2009. View Article : Google Scholar : PubMed/NCBI
34	Yang J, Li J, Lu J, Zhang Y, Zhu Z and Wan H: Synergistic protective effect of astragaloside IV-tetramethylpyrazine against cerebral ischemic-reperfusion injury induced by transient focal ischemia. J Ethnopharmacol. 140:64–72. 2012. View Article : Google Scholar
35	Yin YY, Li WP, Gong HL, Zhu FF, Li WZ and Wu GC: Protective effect of astragaloside on focal cerebral ischemia/reperfusion injury in rats. Am J Chin Med. 38:517–527. 2010. View Article : Google Scholar : PubMed/NCBI
36	Li M, Ma RN, Li LH, Qu YZ and Gao GD: Astragaloside IV reduces cerebral edema post-ischemia/reperfusion correlating the suppression of MMP-9 and AQP4. Eur J Pharmacol. 715:189–195. 2013. View Article : Google Scholar : PubMed/NCBI
37	Li M, Qu YZ, Zhao ZW, Wu SX, Liu YY, Wei XY, Gao L and Gao GD: Astragaloside IV protects against focal cerebral ischemia/reperfusion injury correlating to suppression of neutrophils adhesion-related molecules. Neurochem Int. 60:458–465. 2012. View Article : Google Scholar : PubMed/NCBI
38	Liu P, Tang YY, Yang XS, Dai J, Yang M, Zhang H, Liu Y, Yan H and Song XY: Validation of a preclinical animal model to assess brain recovery after acute stroke. Eur J Pharmacol. 835:75–81. 2018. View Article : Google Scholar : PubMed/NCBI
39	Longa EZ, Weinstein PR, Carlson S and Cummins R: Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 20:84–91. 1989. View Article : Google Scholar : PubMed/NCBI
40	Zhou P, Lu S, Luo Y, Wang S, Yang K, Zhai Y, Sun G and Sun X: Attenuation of TNF-α-induced inflammatory injury in endothelial cells by ginsenoside Rb1 via inhibiting NF-κB, JNK and p38 signaling pathways. Front Pharmacol. 8:4642017. View Article : Google Scholar
41	Peng L, Yin J, Wang S, Ge M, Han Z, Wang Y, Zhang M, Xie L and Li Y: TGF-β2/Smad3 signaling pathway activation through enhancing VEGF and CD34 ameliorates cerebral ischemia/reperfusion injury after isoflurane post-conditioning in rats. Neurochem Res. 44:2606–2618. 2019. View Article : Google Scholar : PubMed/NCBI
42	Wang Y, Li B and Zhang X: Scutellaria barbata D: Don (SBD) protects oxygen glucose deprivation/reperfusion-induced injuries of PC12 cells by up-regulating Nrf2. Artif Cells Nanomed Biotechnol. 47:1797–1807. 2019. View Article : Google Scholar : PubMed/NCBI
43	Zhao DY, Yu DD, Ren L and Bi GR: Ligustilide protects PC12 cells from oxygen-glucose deprivation/reoxygenation-induced apoptosis via the LKB1-AMPK-mTOR signaling pathway. Neural Regen Res. 15:473–481. 2020. View Article : Google Scholar
44	Meng X, Xie W, Xu Q, Liang T, Xu X, Sun G and Sun X: Neuroprotective effects of radix scrophulariae on cerebral ischemia and reperfusion injury via MAPK pathways. Molecules. 23:24012018. View Article : Google Scholar :
45	Li ZR, Yang L, Zhen J, Zhao Y and Lu ZN: Nobiletin protects PC12 cells from ERS-induced apoptosis in OGD/R injury via activation of the PI3K/AKT pathway. Exp Ther Med. 16:1470–1476. 2018.PubMed/NCBI
46	Wang G, Wang T, Zhang Y, Li F, Yu B and Kou J: Schizandrin protects against OGD/R-induced neuronal injury by suppressing autophagy: Involvement of the AMPK/mTOR pathway. Molecules. 24:36242019. View Article : Google Scholar :
47	Donnan GA, Fisher M, Macleod M and Davis SM: Stroke. Lancet. 371:1612–1623. 2008. View Article : Google Scholar : PubMed/NCBI
48	Georgiopoulos G, Ntaios G, Stamatelopoulos K, Manios E, Korompoki E, Vemmou E, Milionis H, Masi S, Lip GYH and Vemmos K: Comparison of risk scores for the prediction of the overall cardiovascular risk in patients with ischemic stroke: The athens stroke registry. J Stroke Cerebrovasc Dis. 28:1044152019. View Article : Google Scholar : PubMed/NCBI
49	Li D and Ai Y: Hydrogen saline suppresses neuronal cell apoptosis and inhibits the p38 mitogenactivated protein kinase-caspase3 signaling pathway following cerebral ischemia-reperfusion injury. Mol Med Rep. 16:5321–5325. 2017. View Article : Google Scholar : PubMed/NCBI
50	Tan XF, Qin T, Li N, Yang YG, Zheng JH, Xie L and Chen MH: High-potassium preconditioning enhances tolerance to focal cerebral ischemia-reperfusion injury through anti-apoptotic effects in male rats. J Neurosci Res. 97:1253–1265. 2019. View Article : Google Scholar : PubMed/NCBI
51	Dirnagl U, Iadecola C and Moskowitz MA: Pathobiology of ischaemic stroke: An integrated view. Trends Neurosci. 22:391–397. 1999. View Article : Google Scholar : PubMed/NCBI
52	Ren Q, Hu Z, Jiang Y, Tan X, Botchway BOA, Amin N, Lin G, Geng Y and Fang M: SIRT1 protects against apoptosis by promoting autophagy in the oxygen glucose deprivation/reperfusion-induced injury. Front Neurol. 10:12892019. View Article : Google Scholar
53	Rogers KV, Dunn CK, Hebert SC and Brown EM: Localization of calcium receptor mRNA in adult rat central nervous system by in situ hybridization. Brain Res. 744:47–56. 1997. View Article : Google Scholar : PubMed/NCBI
54	Ruat M, Molliver ME, Snowman AM and Snyder SH: Calcium sensing receptor: Molecular cloning in rat and localization to nerve terminals. Proc Natl Acad Sci USA. 92:3161–3165. 1995. View Article : Google Scholar : PubMed/NCBI
55	Bandyopadhyay S, Tfelt-Hansen J and Chattopadhyay N: Diverse roles of extracellular calcium-sensing receptor in the central nervous system. J Neurosci Res. 88:2073–2082. 2010. View Article : Google Scholar : PubMed/NCBI
56	Ruat M and Traiffort E: Roles of the calcium sensing receptor in the central nervous system. Best Pract Res Clin Endocrinol Metab. 27:429–442. 2013. View Article : Google Scholar : PubMed/NCBI
57	Sun YH, Li YQ, Feng SL, Li BX, Pan ZW, Xu CQ, Li TT and Yang BF: Calcium-sensing receptor activation contributed to apoptosis stimulates TRPC6 channel in rat neonatal ventricular myocytes. Biochem Biophys Res Commun. 394:955–961. 2010. View Article : Google Scholar : PubMed/NCBI
58	Tfelt-Hansen J, Hansen JL, Smajilovic S, Terwilliger EF, Haunso S and Sheikh SP: Calcium receptor is functionally expressed in rat neonatal ventricular cardiomyocytes. Am J Physiol Heart Circ Physiol. 290:H1165–H1171. 2006. View Article : Google Scholar
59	Zhang G, Zhang T, Wu L, Zhou X, Gu J, Li C, Liu W, Long C, Yang X, Shan L, et al: Neuroprotective effect and mechanism of action of tetramethylpyrazine nitrone for ischemic stroke therapy. Neuromolecular Med. 20:97–111. 2018. View Article : Google Scholar : PubMed/NCBI
60	Lončarević-Vasiljković N, Milanović D, Pešić V, Tešić V, Brkić M, Lazić D, Avramović V and Kanazir S: Dietary restriction suppresses apoptotic cell death, promotes Bcl-2 and Bcl-xl mRNA expression and increases the Bcl-2/Bax protein ratio in the rat cortex after cortical injury. Neurochem Int. 96:69–76. 2016. View Article : Google Scholar
61	Zheng T, Jiang H, Jin R, Zhao Y, Bai Y, Xu H and Chen Y: Ginsenoside Rg1 attenuates protein aggregation and inflammatory response following cerebral ischemia and reperfusion injury. Eur J Pharmacol. 853:65–73. 2019. View Article : Google Scholar : PubMed/NCBI
62	Yu H, Liu P, Tang H, Jing J, Lv X, Chen L, Jiang L, Xu J and Li J: Oleuropein, a natural extract from plants, offers neuroprotection in focal cerebral ischemia/reperfusion injury in mice. Eur J Pharmacol. 775:113–119. 2016. View Article : Google Scholar : PubMed/NCBI
63	Wang Y, Luo J and Li SY: Nano-curcumin simultaneously protects the blood-brain barrier and reduces M1 microglial activation during cerebral ischemia-reperfusion injury. ACS Appl Mater Interfaces. 11:3763–3770. 2019. View Article : Google Scholar : PubMed/NCBI
64	Hou Y, Wang J and Feng J: The neuroprotective effects of curcumin are associated with the regulation of the reciprocal function between autophagy and HIF-1α in cerebral ischemia-reperfusion injury. Drug Des Devel Ther. 13:1135–1144. 2019. View Article : Google Scholar :