Paeoniflorin protects PC12 cells from oxygen-glucose deprivation/reoxygenation-induced injury via activating JAK2/STAT3 signaling

Zhang,Zhuo; Yang,Weimin

doi:10.3892/etm.2021.10004

Paeoniflorin protects PC12 cells from oxygen-glucose deprivation/reoxygenation-induced injury via activating JAK2/STAT3 signaling

Authors:
- Zhuo Zhang
- Weimin Yang
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Affiliations: Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450000, P.R. China
Published online on: March 29, 2021 https://doi.org/10.3892/etm.2021.10004
Article Number: 572
Copyright: © Zhang 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 the most common type of stroke, and it has become a major health issue as it is characterized by high mortality and morbidity rates. Paeoniflorin (PF) is a natural compound and the main active ingredient of Radix Paeoniae. The aim of the present study was to investigate the role of PF in oxygen‑glucose deprivation/reoxygenation (OGD/R)‑induced injury of PC12 cells and its association with the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway. An in vitro model of OGD/R injury was established in PC12 cells. Subsequently, Cell Counting Kit‑8 assay and ELISA were used to evaluate cell viability and the secretion of inflammatory factors, respectively, in PC12 cells subjected to OGD/R and treated with PF. The levels of oxidative stress indicators and inflammatory factors were measured using corresponding commercial kits. In addition, the apoptosis rate of PC12 cells subjected to OGD/R and treated with PF was determined by flow cytometry, and the expression of apoptosis‑related proteins was analyzed by western blotting. Additionally, the expression levels of JAK2/STAT3 pathway‑related proteins were also evaluated. The cell viability, levels of oxidative stress, inflammation and apoptosis were also measured in OGD/R‑induced PC12 cell injury models following co‑treatment of cells with PF and FLLL32, a specific inhibitor of JAK2/STAT3 signaling. Cell viability was reduced, while oxidative stress and inflammation were increased after OGD/R‑induced injury. However, the treatment of cells with PF significantly enhanced cell viability, and alleviated oxidative stress, inflammation and apoptosis of OGD/R‑treated PC12 cells. Furthermore, PF activated the JAK2/STAT3 signaling pathway. Following FLLL32 intervention, the effects of PF on oxidative stress, inflammation and apoptosis of OGD/R‑treated PC12 cells were reversed. In conclusion, the findings of the present study suggested that PF may protect PC12 cells from OGD/R‑induced injury via activating the JAK2/STAT3 signaling pathway, thus providing novel insight into the mechanism through which PF may alleviate ischemic stroke and indicating a potential strategy for ischemic stroke treatment.

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1	Martini ML, Neifert SN, Lara-Reyna JJ, Shuman WH, Ladner TR, Hardigan TH, Fifi JT, Mocco J and Yaeger KA: Trials in thrombectomy for acute ischemic stroke: Describing the state of clinical research in the field. Clin Neurol Neurosurg. 200(106360)2021.PubMed/NCBI View Article : Google Scholar
2	Chen H, Yoshioka H, Kim GS, Jung JE, Okami N, Sakata H, Maier CM, Narasimhan P, Goeders CE and Chan PH: Oxidative stress in ischemic brain damage: Mechanisms of cell death and potential molecular targets for neuroprotection. Antioxid Redox Signal. 14:1505–1517. 2011.PubMed/NCBI View Article : Google Scholar
3	Bavarsad K, Barreto GE, Hadjzadeh MA and Sahebkar A: Protective effects of curcumin against ischemia-reperfusion injury in the nervous system. Mol Neurobiol. 56:1391–1404. 2019.PubMed/NCBI View Article : Google Scholar
4	Chen A, Wang H, Zhang Y, Wang X, Yu L, Xu W, Xu W and Lin Y: Paeoniflorin exerts neuroprotective effects against glutamate induced PC12 cellular cytotoxicity by inhibiting apoptosis. Int J Mol Med. 40:825–833. 2017.PubMed/NCBI View Article : Google Scholar
5	Zhai A, Zhang Z and Kong X: Paeoniflorin alleviates H₂O₂-induced oxidative injury through down-regulation of microRNA-135a in HT-22 cells. Neurochem Res. 44:2821–2831. 2019.PubMed/NCBI View Article : Google Scholar
6	Wen Z, Hou W, Wu W, Zhao Y, Dong X, Bai X, Peng L and Song L: 6'-O-Galloylpaeoniflorin attenuates cerebral ischemia reperfusion-induced neuroinflammation and oxidative stress via PI3K/Akt/Nrf2 activation. Oxid Med Cell Longev. 2018(8678267)2018.PubMed/NCBI View Article : Google Scholar
7	Liu H, Yu C, Xu T, Zhang X and Dong M: Synergistic protective effect of paeoniflorin and β-ecdysterone against rotenone-induced neurotoxicity in PC12 cells. Apoptosis. 21:1354–1365. 2016.PubMed/NCBI View Article : Google Scholar
8	Shio MT, Christian JG, Jung JY, Chang KP and Olivier M: PKC/ROS-mediated NLRP3 inflammasome activation is attenuated by leishmania zinc-metalloprotease during Infection. PLoS Negl Trop Dis. 9(e0003868)2015.PubMed/NCBI View Article : Google Scholar
9	Li J, Ji X, Zhang J, Shi G, Zhu X and Wang K: Paeoniflorin attenuates Aβ25-35-induced neurotoxicity in PC12 cells by preventing mitochondrial dysfunction. Folia Neuropathol. 52:285–290. 2014.PubMed/NCBI
10	Wang D, Wong HK, Feng YB and Zhang ZJ: Paeoniflorin, a natural neuroprotective agent, modulates multiple anti-apoptotic and pro-apoptotic pathways in differentiated PC12 cells. Cell Mol Neurobiol. 33:521–529. 2013.PubMed/NCBI View Article : Google Scholar
11	Hou Y, Wang K, Wan W, Cheng Y, Pu X and Ye X: Resveratrol provides neuroprotection by regulating the JAK2/STAT3/PI3K/AKT/mTOR pathway after stroke in rats. Genes Dis. 5:245–255. 2018.PubMed/NCBI View Article : Google Scholar
12	de Couto G, Liu W, Tseliou E, Sun B, Makkar N, Kanazawa H, Arditi M and Marbán E: Macrophages mediate cardioprotective cellular postconditioning in acute myocardial infarction. J Clin Invest. 125:3147–3162. 2015.PubMed/NCBI View Article : Google Scholar
13	Li X, Wang Y, Wang K and Wu Y: Renal protective effect of Paeoniflorin by inhibition of JAK2/STAT3 signaling pathway in diabetic mice. Biosci Trends. 12:168–176. 2018.PubMed/NCBI View Article : Google Scholar
14	Yang B, Zang LE, Cui JW, Zhang MY, Ma X and Wei LL: Melatonin plays a protective role by regulating miR-26a-5p-NRSF and JAK2-STAT3 pathway to improve autophagy, inflammation and oxidative stress of cerebral ischemia-reperfusion injury. Drug Des Devel Ther. 14:3177–3188. 2020.PubMed/NCBI View Article : Google Scholar
15	Yuan J, Zeng L, Sun Y, Wang N, Sun Q, Cheng Z and Wang Y: SH2B1 protects against OGD/R induced apoptosis in PC12 cells via activation of the JAK2/STAT3 signaling pathway. Mol Med Rep. 18:2613–2620. 2018.PubMed/NCBI View Article : Google Scholar
16	Zhang LL, Wei W, Wang NP, Wang QT, Chen JY, Chen Y, Wu H and Hu XY: Paeoniflorin suppresses inflammatory mediator production and regulates G protein-coupled signaling in fibroblast-like synoviocytes of collagen induced arthritic rats. Inflamm Res. 57:388–395. 2008.PubMed/NCBI View Article : Google Scholar
17	Ling L, Tong J and Zeng L: Paeoniflorin improves acute lung injury in sepsis by activating Nrf2/Keap1 signaling pathway. Sichuan Da Xue Xue Bao Yi Xue Ban. 51:664–669. 2020.PubMed/NCBI View Article : Google Scholar : (In Chinese).
18	Zhou H, Bian D, Jiao X, Wei Z, Zhang H, Xia Y, He Y and Dai Y: Paeoniflorin protects against lipopolysaccharide-induced acute lung injury in mice by alleviating inflammatory cell infiltration and microvascular permeability. Inflamm Res. 60:981–990. 2011.PubMed/NCBI View Article : Google Scholar
19	Jiang WL, Chen XG, Zhu HB, Gao YB, Tian JW and Fu FH: Paeoniflorin inhibits systemic inflammation and improves survival in experimental sepsis. Basic Clin Pharmacol Toxicol. 105:64–71. 2009.PubMed/NCBI View Article : Google Scholar
20	Wei S, Ma X, Niu M, Wang R, Yang T, Wang D, Wen J, Li H and Zhao Y: Mechanism of paeoniflorin in the treatment of bile duct ligation-induced cholestatic liver injury using integrated metabolomics and network pharmacology. Front Pharmacol. 11(586806)2020.PubMed/NCBI View Article : Google Scholar
21	Poprac P, Jomova K, Simunkova M, Kollar V, Rhodes CJ and Valko M: Targeting free radicals in oxidative stress-related human diseases. Trends Pharmacol Sci. 38:592–607. 2017.PubMed/NCBI View Article : Google Scholar
22	Chen S, Chen H, Du Q and Shen J: Targeting myeloperoxidase (MPO) mediated oxidative stress and inflammation for reducing brain ischemia injury: Potential application of natural compounds. Front Physiol. 11(433)2020.PubMed/NCBI View Article : Google Scholar
23	Yu G, Liang Y, Huang Z, Jones DW, Pritchard KA Jr and Zhang H: Erratum to: Inhibition of myeloperoxidase oxidant production by N-acetyl lysyltyrosylcysteine amide reduces brain damage in a murine model of stroke. J Neuroinflammation. 13(166)2016.PubMed/NCBI View Article : Google Scholar
24	Briyal S, Gulati K and Gulati A: Repeated administration of exendin-4 reduces focal cerebral ischemia-induced infarction in rats. Brain Res. 1427:23–34. 2012.PubMed/NCBI View Article : Google Scholar
25	Zhao L, Xu J, Wang Q, Qian Z, Feng W, Yin X and Fang Y: Protective effect of rhGLP-1 (7-36) on brain ischemia/reperfusion damage in diabetic rats. Brain Res. 1602:153–159. 2015.PubMed/NCBI View Article : Google Scholar
26	Fang Y, Liu X, Zhao L, Wei Z, Jiang D, Shao H, Zang Y, Xu J, Wang Q, Liu Y, et al: RhGLP-1 (7-36) protects diabetic rats against cerebral ischemia-reperfusion injury via up-regulating expression of Nrf2/HO-1 and increasing the activities of SOD. Korean J Physiol Pharmacol. 21:475–485. 2017.PubMed/NCBI View Article : Google Scholar
27	Dai X, Wang LW, Jia XY, Chang Y, Wu HX, Wang C and Wei W: Paeoniflorin regulates the function of human peripheral blood mononuclear cells stimulated by rhIL-1β by up-regulating Treg expression. Immunopharmacol Immunotoxicol. 37:252–257. 2015.PubMed/NCBI View Article : Google Scholar
28	Jin L, Zhang LM, Xie KQ, Ye Y and Feng L: Paeoniflorin suppresses the expression of intercellular adhesion molecule-1 (ICAM-1) in endotoxin-treated human monocytic cells. Br J Pharmacol. 164:694–703. 2011.PubMed/NCBI View Article : Google Scholar
29	Zhai T, Sun Y, Li H, Zhang J, Huo R, Li H, Shen B and Li N: Unique immunomodulatory effect of paeoniflorin on type I and II macrophages activities. J Pharmacol Sci. 130:143–150. 2016.PubMed/NCBI View Article : Google Scholar
30	Shi D, Wang Q, Zheng H, Li D, Shen Y, Fu H, Li T, Mei H, Lu G, Qiu Y, et al: Paeoniflorin suppresses IL-6/Stat3 pathway via upregulation of Socs3 in dendritic cells in response to 1-chloro-2,4-dinitrobenze. Int Immunopharmacol. 38:45–53. 2016.PubMed/NCBI View Article : Google Scholar
31	Adams JM and Cory S: The Bcl-2 protein family: Arbiters of cell survival. Science. 281:1322–1326. 1998.PubMed/NCBI View Article : Google Scholar
32	Shyu WC, Lin SZ, Chiang MF, Chen DC, Su CY, Wang HJ, Liu RS, Tsai CH and Li H: Secretoneurin promotes neuroprotection and neuronal plasticity via the Jak2/Stat3 pathway in murine models of stroke. J Clin Invest. 118:133–148. 2008.PubMed/NCBI View Article : Google Scholar
33	Hossain E, Li Y and Anand-Srivastava MB: Role of JAK2/STAT3 pathway in angiotensin II-induced enhanced expression of Gialpha proteins and hyperproliferation of aortic vascular smooth muscle cells. Can J Physiol Pharmacol. 99:237–246. 2021.PubMed/NCBI View Article : Google Scholar
34	El-Ghafar OAMA, Helal GK and Abo-Youssef AM: Apixaban exhibits anti-arthritic effects by inhibiting activated factor X-mediated JAK2/STAT3 and MAPK phosphorylation pathways. Inflammopharmacology. 28:1253–1267. 2020.PubMed/NCBI View Article : Google Scholar
35	Pan S, Chen Y, Zhang X and Xie Y: The JAK2/STAT3 pathway is involved in dexmedetomidine-induced myocardial protection in rats undergoing cardiopulmonary bypass. Ann Transl Med. 8(483)2020.PubMed/NCBI View Article : Google Scholar
36	Wu QY, Ma MM, Fu L, Zhu YY, Liu Y, Cao J, Zhou P, Li ZY, Zeng LY, Li F, et al: Roles of germline JAK2 activation mutation JAK2 V625F in the pathology of myeloproliferative neoplasms. Int J Biol Macromol. 116:1064–1073. 2018.PubMed/NCBI View Article : Google Scholar
37	Leong PL, Andrews GA, Johnson DE, Dyer KF, Xi S, Mai JC, Robbins PD, Gadiparthi S, Burke NA, Watkins SF, et al: Targeted inhibition of Stat3 with a decoy oligonucleotide abrogates head and neck cancer cell growth. Proc Natl Acad Sci USA. 100:4138–4143. 2003.PubMed/NCBI View Article : Google Scholar
38	Li L, Li H and Li M: Curcumin protects against cerebral ischemia-reperfusion injury by activating JAK2/STAT3 signaling pathway in rats. Int J Clin Exp Med. 8:14985–14991. 2015.PubMed/NCBI
39	Li L, Sun L, Qiu Y, Zhu W, Hu K and Mao J: Protective effect of stachydrine against cerebral ischemia-reperfusion injury by reducing inflammation and apoptosis through P65 and JAK2/STAT3 signaling pathway. Front Pharmacol. 11(64)2020.PubMed/NCBI View Article : Google Scholar