Hydroxysafflor Yellow A mitigated myocardial ischemia/reperfusion injury by inhibiting the activation of the JAK2/STAT1 pathway

Zhou,Donglai; Ding,Tingting; Ni,Bin; Jing,Yunyan; Liu,Shanxin

doi:10.3892/ijmm.2019.4230

Hydroxysafflor Yellow A mitigated myocardial ischemia/reperfusion injury by inhibiting the activation of the JAK2/STAT1 pathway

Authors:
- Donglai Zhou
- Tingting Ding
- Bin Ni
- Yunyan Jing
- Shanxin Liu
View Affiliations

Affiliations: Department of Cardiology, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang 310015, P.R. China
Published online on: June 5, 2019 https://doi.org/10.3892/ijmm.2019.4230
Pages: 405-416
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hydroxysafflor Yellow A (HSYA) may reduce ischemia/reperfusion (I/R) injury. However, the underlying molecular mechanisms remain unclear. The present study explored the effect and the mechanisms of HSYA on myocardial injury in vivo and in vitro. Myocardial infarct size was assessed by Evans blue/2,3,5‑triphenyltetrazoliumchloride staining. Levels of cardiac troponin I (cTnI), interleukin‑6 (IL‑6), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured using commercial kits. Alteration of mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) generation was determined by fluorescent signals. Apoptosis was detected by terminal deoxynucleotidyl‑transferase‑mediated dUTP nick‑end labeling staining, flow cytometry assay and caspase‑3 activity. Expression levels of the apoptosis‑associated proteins were detected by reverse transcription quantitative polymerase chain reaction and western blot analysis. In vivo, animals treated with HSYA presented less severe myocardial injury and decreased janus kinase 2 (JAK2)/signal transducer and activator of transcription 1 (STAT1) activity, improved antioxidant capacity and decreased apoptosis. In vitro, compared with the hypoxia (H)/reoxygenation (R) + HSYA group, AG490 and S1491 treatment decreased the releases of cTnI, IL‑6 and LDH and enhanced the resistance to oxidative stress by maintaining MMP and decreasing ROS generation. In addition, AG490 and S1491 were also identified to alleviate the H/R‑induced apoptosis by inhibiting caspase 3 activity and modulating the expression levels of cleaved caspase‑3, tumor necrosis factor receptor superfamily member 6 (Fas), Fas ligand, B‑cell lymphoma 2 (Bcl‑2) and Bcl‑2‑associated X protein. These data suggested that inactivation of the JAK2/STAT1 pathway strengthened the HSYA‑induced protective effect in H/R‑induced myocardial injury. In conclusion, the treatment of HSYA was effective in decreasing IR‑induced myocardial injury, and this may be largely dependent on the JAK2/STAT1 pathway. Therefore, the present study provided a potential strategy to prevent myocardial I/R injury.

View Figures

View References

1	Bainey KR and Armstrong PW: Clinical perspectives on reperfusion injury in acute myocardial infarction. Am Heart J. 167:637–645. 2014. View Article : Google Scholar : PubMed/NCBI
2	Maxwell SR and Lip GY: Reperfusion injury: A review of the pathophysiology, clinical manifestations and therapeutic options. Int J Cardiol. 58:95–117. 1997. View Article : Google Scholar : PubMed/NCBI
3	Heusch G, Boengler K and Schulz R: Inhibition of mitochondrial permeability transition pore opening: The Holy Grail of cardio-protection. Basic Res Cardiol. 105:151–154. 2010. View Article : Google Scholar : PubMed/NCBI
4	GBD 2015 Disease and Injury Incidence and Prevalence Collaborators: Global, regional, and national incidence, prevalence, and years lived with disability for 310 diseases and injuries, 1990-2015: A systematic analysis for the Global Burden of Disease Study, 2015. Lancet. 388:1545–1602. 2016. View Article : Google Scholar
5	Yellon DM and Hausenloy DJ: Myocardial reperfusion injury. N Engl J Med. 357:1121–1135. 2007. View Article : Google Scholar : PubMed/NCBI
6	Xiong J, Xue FS, Yuan YJ, Wang Q, Liao X and Wang WL: Cholinergic anti-inflammatory pathway: A possible approach to protect against myocardial ischemia reperfusion injury. Chin Med J (Engl). 123:2720–2726. 2010.
7	Naidu BV, Farivar AS, Woolley SM, Grainger D, Verrier ED and Mulligan MS: Novel broad-spectrum chemokine inhibitor protects against lung ischemia-reperfusion injury. J Heart Lung Transplant. 23:128–134. 2004. View Article : Google Scholar : PubMed/NCBI
8	Olivetti G, Quaini F, Sala R, Lagrasta C, Corradi D, Bonacina E, Gambert SR, Cigola E and Anversa P: Acute myocardial infarction in humans is associated with activation of programmed myocyte cell death in the surviving portion of the heart. J Mol Cell Cardiol. 28:2005–2016. 1996. View Article : Google Scholar : PubMed/NCBI
9	Zhao ZQ, Nakamura M, Wang NP, Wilcox JN, Shearer S, Ronson RS, Guyton RA and Vinten-Johansen J: Reperfusion induces myocardial apoptotic cell death. Cardiovasc Res. 45:651–660. 2000. View Article : Google Scholar : PubMed/NCBI
10	Orogo AM and Gustafsson AB: Cell death in the myocardium: My heart won't go on. IUBMB Life. 65:651–656. 2013. View Article : Google Scholar : PubMed/NCBI
11	Stephanou A, Brar B, Liao Z, Scarabelli T, Knight RA and Latchman DS: Distinct initiator caspases are required for the induction of apoptosis in cardiac myocytes during ischaemia versus reperfusion injury. Cell Death Differ. 8:434–435. 2001. View Article : Google Scholar : PubMed/NCBI
12	Scarabelli TM, Stephanou A, Pasini E, Comini L, Raddino R, Knight RA and Latchman DS: Different signaling pathways induce apoptosis in endothelial cells and cardiac myocytes during ischemia/reperfusion injury. Circ Res. 90:745–748. 2002. View Article : Google Scholar : PubMed/NCBI
13	Scarabelli T, Stephanou A, Rayment N, Pasini E, Comini L, Curello S, Ferrari R, Knight R and Latchman D: Apoptosis of endothelial cells precedes myocyte cell apoptosis in ischemia/reperfusion injury. Circulation. 104:253–256. 2001. View Article : Google Scholar : PubMed/NCBI
14	Jeremias I, Kupatt C, Martin-Villalba A, Habazettl H, Schenkel J, Boekstegers P and Debatin KM: Involvement of CD95/Apo1/Fas in cell death after myocardial ischemia. Circulation. 102:915–920. 2000. View Article : Google Scholar : PubMed/NCBI
15	Jin M, Li JR and Wu W: Study on the antioxidative effect of safflor yellow. Zhongguo Zhong Yao Za Zhi. 29:447–449. 2004.In Chinese.
16	Wei X, Liu H, Sun X, Fu F, Zhang X, Wang J, An J and Ding H: Hydroxysafflor yellow A protects rat brains against ischemia-reperfusion injury by antioxidant action. Neurosci Lett. 386:58–62. 2005. View Article : Google Scholar : PubMed/NCBI
17	Darnell JE Jr: STATs and gene regulation. Science. 277:1630–1635. 1997. View Article : Google Scholar : PubMed/NCBI
18	Imada K and Leonard WJ: The Jak-STAT pathway. Mol Immunol. 37:1–11. 2000. View Article : Google Scholar : PubMed/NCBI
19	Negoro S, Kunisada K, Tone E, Funamoto M, Oh H, Kishimoto T and Yamauchi-Takihara K: Activation of JAK/STAT pathway transduces cytoprotective signal in rat acute myocardial infarction. Cardiovasc Res. 47:797–805. 2000. View Article : Google Scholar : PubMed/NCBI
20	Das A, Salloum FN, Durrant D, Ockaili R and Kukreja RC: Rapamycin protects against myocardial ischemia-reperfusion injury through JAK2-STAT3 signaling pathway. J Mol Cell Cardiol. 53:858–869. 2012. View Article : Google Scholar : PubMed/NCBI
21	Battle TE and Frank DA: The role of STATs in apoptosis. Curr Mol Med. 2:381–392. 2002. View Article : Google Scholar : PubMed/NCBI
22	Boengler K, Hilfiker-Kleiner D, Heusch G and Schulz R: Inhibition of permeability transition pore opening by mitochondrial STAT3 and its role in myocardial ischemia/reperfusion. Basic Res Cardiol. 105:771–785. 2010. View Article : Google Scholar : PubMed/NCBI
23	Heusch G, Musiolik J, Gedik N and Skyschally A: Mitochondrial STAT3 activation and cardioprotection by ischemic postconditioning in pigs with regional myocardial ischemia/reperfusion. Circ Res. 109:1302–1308. 2011. View Article : Google Scholar : PubMed/NCBI
24	Yang N, Luo M, Li R, Huang Y, Zhang R, Wu Q, Wang F, Li Y and Yu X: Blockage of JAK/STAT signalling attenuates renal ischaemia-reperfusion injury in rat. Nephrol Dial Transplant. 23:91–100. 2008. View Article : Google Scholar
25	Stephanou A, Brar BK, Knight RA and Latchman DS: Opposing actions of STAT-1 and STAT-3 on the Bcl-2 and Bcl-x promoters. Cell Death Differ. 7:329–330. 2000. View Article : Google Scholar : PubMed/NCBI
26	Stephanou A, Scarabelli TM, Townsend PA, Bell R, Yellon D, Knight RA and Latchman DS: The carboxyl-terminal activation domain of the STAT-1 transcription factor enhances ischemia/reperfusion-induced apoptosis in cardiac myocytes. FASEB J. 16:1841–1843. 2002. View Article : Google Scholar : PubMed/NCBI
27	National Research Council (US) Committee for the Update of the Guide for the Care and Use of Laboratory Animals: Guide for the Care and Use of Laboratory Animals. 8th edition. National Academies Press (US); Washington, DC: 2011
28	Zhu HB, Zhang L, Wang ZH, Tian JW, Fu FH, Liu K and Li CL: Therapeutic effects of hydroxysafflor yellow A on focal cerebral ischemic injury in rats and its primary mechanisms. J Asian Nat Prod Res. 7:607–613. 2005. View Article : Google Scholar : PubMed/NCBI
29	Liu SX, Zhang Y, Wang YF, Li XC, Xiang MX, Bian C and Chen P: Upregulation of heme oxygenase-1 expression by hydroxysafflor yellow A conferring protection from anoxia/reoxygenation-induced apoptosis in H9c2 cardiomyocytes. Int J Cardiol. 160:95–101. 2012. View Article : Google Scholar
30	Wu YX, Gao CZ, Fan KL, Yang LM and Mei XF: STAT1 inhibitor alleviates spinal cord injury by decreasing apoptosis. Genet Mol Res. 15:2016.
31	Gorina R, Petegnief V, Chamorro A and Planas AM: AG490 prevents cell death after exposure of rat astrocytes to hydrogen peroxide or proinflammatory cytokines: Involvement of the Jak2/STAT pathway. J Neurochem. 92:505–518. 2005. View Article : Google Scholar : PubMed/NCBI
32	Smith CC, Dixon RA, Wynne AM, Theodorou L, Ong SG, Subrayan S, Davidson SM, Hausenloy DJ and Yellon DM: Leptin-induced cardioprotection involves JAK/STAT signaling that may be linked to the mitochondrial permeability transition pore. Am J Physiol Heart Circ Physiol. 299:H1265–H1270. 2010. View Article : Google Scholar : PubMed/NCBI
33	Sachdeva J, Dai W, Gerczuk PZ and Kloner RA: Combined remote perconditioning and postconditioning failed to attenuate infarct size and contractile dysfunction in a rat model of coronary artery occlusion. J Cardiovasc Pharmacol Ther. 19:567–573. 2014. View Article : Google Scholar : PubMed/NCBI
34	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
35	Liu F, Wei Y, Yang XZ, Li FG, Hu J and Cheng RF: Hypotensive effects of safflower yellow in spontaneously hypertensive rats and influence on plasma renin activity and angiotensin II level. Yao Xue Xue Bao. 27:785–787. 1992.In Chinese.
36	Ji DB, Zhang LY, Li CL, Ye J and Zhu HB: Effect of Hydroxysafflor yellow A on human umbilical vein endothelial cells under hypoxia. Vascul Pharmacol. 50:137–145. 2009. View Article : Google Scholar
37	Panteghini M, Cuccia C, Bonetti G, Giubbini R, Pagani F and Bonini E: Single-point cardiac troponin T at coronary care unit discharge after myocardial infarction correlates with infarct size and ejection fraction. Clin Chem. 48:1432–1436. 2002.PubMed/NCBI
38	Kowalewski M, Urban M, Mroczko B and Szmitkowski M: Proinflammatory cytokines (IL-6, TNF-alpha) and cardiac troponin I (cTnI) in serum of young people with ventricular arrhythmias. Pol Arch Med Wewn. 108:647–651. 2002.In Polish. PubMed/NCBI
39	Das DK, Engelman RM, Rousou JA, Breyer RH, Otani H and Lemeshow S: Pathophysiology of superoxide radical as potential mediator of reperfusion injury in pig heart. Basic Res Cardiol. 81:155–166. 1986. View Article : Google Scholar : PubMed/NCBI
40	Wu J, Hecker JG and Chiamvimonvat N: Antioxidant enzyme gene transfer for ischemic diseases. Adv Drug Deliv Rev. 61:351–363. 2009. View Article : Google Scholar : PubMed/NCBI
41	Taiwo T and Goldstein S: Drug use and its association with deviant behaviour among rural adolescent students in South Africa. East Afr Med J. 83:500–506. 2006.
42	Scarabelli TM, Knight R, Stephanou A, Townsend P, Chen-Scarabelli C, Lawrence K, Gottlieb R, Latchman D and Narula J: Clinical implications of apoptosis in ischemic myocardium. Curr Probl Cardiol. 31:181–264. 2006. View Article : Google Scholar : PubMed/NCBI
43	Salvesen GS: Caspases and apoptosis. Essays Biochem. 38:9–19. 2002. View Article : Google Scholar : PubMed/NCBI
44	Barry SP, Townsend PA, Latchman DS and Stephanou A: Role of the JAK-STAT pathway in myocardial injury. Trends Mol Med. 13:82–89. 2007. View Article : Google Scholar
45	Shinmura K, Tang XL, Wang Y, Xuan YT, Liu SQ, Takano H, Bhatnagar A and Bolli R: Cyclooxygenase-2 mediates the cardio-protective effects of the late phase of ischemic preconditioning in conscious rabbits. Proc Natl Acad Sci USA. 97:10197–10202. 2000. View Article : Google Scholar
46	Fuglesteg BN, Suleman N, Tiron C, Kanhema T, Lacerda L, Andreasen TV, Sack MN, Jonassen AK, Mjøs OD, Opie LH and Lecour S: Signal transducer and activator of transcription 3 is involved in the cardioprotective signalling pathway activated by insulin therapy at reperfusion. Basic Res Cardiol. 103:444–453. 2008. View Article : Google Scholar : PubMed/NCBI
47	Mascareno E, El-Shafei M, Maulik N, Sato M, Guo Y, Das DK and Siddiqui MA: JAK/STAT signaling is associated with cardiac dysfunction during ischemia and reperfusion. Circulation. 104:325–329. 2001. View Article : Google Scholar : PubMed/NCBI
48	Simon AR, Rai U, Fanburg BL and Cochran BH: Activation of the JAK-STAT pathway by reactive oxygen species. Am J Physiol. 275:C1640–C1652. 1998. View Article : Google Scholar : PubMed/NCBI
49	Takagi Y, Harada J, Chiarugi A and Moskowitz MA: STAT1 is activated in neurons after ischemia and contributes to ischemic brain injury. J Cereb Blood Flow Metab. 22:1311–1318. 2002. View Article : Google Scholar : PubMed/NCBI
50	Sinha K, Das J, Pal PB and Sil PC: Oxidative stress: The mitochondria-dependent and mitochondria-independent pathways of apoptosis. Arch Toxicol. 87:1157–1180. 2013. View Article : Google Scholar : PubMed/NCBI
51	Wang J, Ouyang C, Chen X, Fu B, Lu Y and Hong Q: Effect of Jak2 kinase inhibition on Stat1 and Stat3 activation and apoptosis of tubular epithelial cells induced by ATP depletion/recovery. J Nephrol. 21:919–923. 2008.PubMed/NCBI
52	Dhingra S, Bagchi AK, Ludke AL, Sharma AK and Singal PK: Akt regulates IL-10 mediated suppression of TNFα-induced cardiomyocyte apoptosis by upregulating Stat3 phosphorylation. PLoS One. 6:e250092011. View Article : Google Scholar
53	Stephanou A, Brar BK, Scarabelli TM, Jonassen AK, Yellon DM, Marber MS, Knight RA and Latchman DS: Ischemia-induced STAT-1 expression and activation play a critical role in cardio-myocyte apoptosis. J Biol Chem. 275:10002–10008. 2000. View Article : Google Scholar : PubMed/NCBI
54	Bolli R, Dawn B and Xuan YT: Role of the JAK-STAT pathway in protection against myocardial ischemia/reperfusion injury. Trends Cardiovasc Med. 13:72–79. 2003. View Article : Google Scholar : PubMed/NCBI
55	Sheng M, Huang Z, Pan L, Yu M, Yi C, Teng L, He L, Gu C, Xu C and Li J: SOCS2 exacerbates myocardial injury induced by ischemia/reperfusion in diabetic mice and H9c2 cells through inhibiting the JAK-STAT-IGF-1 pathway. Life Sci. 188:101–109. 2017. View Article : Google Scholar : PubMed/NCBI
56	Yamaura G, Turoczi T, Yamamoto F, Siddqui MA, Maulik N and Das DK: STAT signaling in ischemic heart: A role of STAT5A in ischemic preconditioning. Am J Physiol Heart Circ Physiol. 285:H476–H482. 2003. View Article : Google Scholar : PubMed/NCBI
57	Jung YY, Lee JH, Nam D, Narula AS, Namjoshi OA, Blough BE, Um JY, Sethi G and Ahn KS: Anti-myeloma effects of icariin are mediated through the attenuation of JAK/STAT3-dependent signaling cascade. Front Pharmacol. 9:5312018. View Article : Google Scholar : PubMed/NCBI
58	Han D, Wei J, Zhang R, Ma W, Shen C, Feng Y, Xia N, Xu D, Cai D, Li Y and Fang W: Hydroxysafflor yellow A alleviates myocardial ischemia/reperfusion in hyperlipidemic animals through the suppression of TLR4 signaling. Sci Rep. 6:353192016. View Article : Google Scholar : PubMed/NCBI
59	Min J and Wei C: Hydroxysafflor yellow A cardioprotection in ischemia-reperfusion (I/R) injury mainly via Akt/hexokinase II independent of ERK/GSK-3β pathway. Biomed Pharmacother. 87:419–426. 2017. View Article : Google Scholar : PubMed/NCBI
60	Xu AB, Liu JG, Zhang J and LI JX: Hydroxysafflor yellow A for ameliorating injury of myocardial ischemia/reperfusion in SD rats. Chin J Evidence Based Cardiovascular Med. 6:733–736. 2014.In Chinese.