Chinese herbal medicine Xinji pill protects the heart from ischemia/reperfusion injury through the Akt/Nrf2 pathway

Yuan,Qiuzhen; Chen,Ruiming; Zheng,Xu; Meng,Maixia; Kao,Yuping; Liu,Junfeng; Gan,Xuefeng; Shi,Minjuan; Fu,Junming; Jiang,Shanshan; Yu,Huiyao

doi:10.3892/mmr.2017.6732

Chinese herbal medicine Xinji pill protects the heart from ischemia/reperfusion injury through the Akt/Nrf2 pathway

Authors:
- Qiuzhen Yuan
- Ruiming Chen
- Xu Zheng
- Maixia Meng
- Yuping Kao
- Junfeng Liu
- Xuefeng Gan
- Minjuan Shi
- Junming Fu
- Shanshan Jiang
- Huiyao Yu
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Affiliations: Department of Pharmacy, Shanxi Hospital of Traditional Chinese Medicine, Xi'an, Shaanxi 710032, P.R. China, Institute of TCM Research, Shaanxi Provincial Academy of Traditional Chinese Medicine, Xianyang, Shaanxi 712046, P.R. China, Department of Pharmacology, Shaanxi Institute for Food and Drug Control, Xi'an, Shaanxi 710000, P.R. China, Scientific Research Department, Shaanxi Hospital of Traditional Chinese Medicine, Xi'an, Shaanxi 710032, P.R. China
Published online on: June 8, 2017 https://doi.org/10.3892/mmr.2017.6732
Pages: 1551-1558

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Abstract

The cardioprotective drugs used for treatment against ischemia/reperfusion (MI/R) injury have been well evaluated and are considered inadequate. The Chinese herbal medicine formula, Xinji pill (XJP) has been used traditionally for the prevention and treatment of ischemic heart diseases for decades. In the present study, the cardioprotective effects of XJP against MI/R injury were assessed in vivo and its possible mechanism was examined. Male Sprague‑Dawley rats were selected for establishing an MI/R model, which was induced by ischemia for 30 min followed by 24 h reperfusion. Drugs and saline were administered intragastrically from day 14 prior to MI/R. Blood samples were collected for biochemical detection. The rats were then sacrificed and cardiac muscle tissues were harvested. The mRNA expression levels of antioxidant genes were measured by reverse transcription‑quantitative polymerase chain reaction and the protein levels were measured by western blotting. Pretreatment with XJP for 14 days protected the heart against I/R‑induced myocardial function disorder, protected against heart injury, as demonstrated by normalized serum levels of lactate dehydrogenase and creatine kinase, and suppressed oxidative stress. XJP markedly upregulated the expression of antioxidant genes, including superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase, and promoted the protein expression of heme oxygenase‑1 and NFE2‑related factor 2 (Nrf2) in the heart tissues. Furthermore, Akt kinase was confirmed to be upstream of Nrf2 in the XJP treatment. LY294002, a speciﬁc inhibitor of Akt, significantly eliminated the cardioprotective effects of XJP. In conclusion, these results demonstrated that XJP exhibited notable cardioprotective properties, in which the Akt/Nrf2 signaling pathway may be involved.

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1	Zhong X, Li X, Qian L, Xu Y, Lu Y, Zhang J, Li N, Zhu X, Ben J, Yang Q and Chen Q: Glycine attenuates myocardial ischemia-reperfusion injury by inhibiting myocardial apoptosis in rats. J Biomed Res. 26:346–354. 2012. View Article : Google Scholar : PubMed/NCBI
2	Piper HM, García-Dorado D and Ovize M: A fresh look at reperfusion injury. Cardiovasc Res. 38:291–300. 1998. View Article : Google Scholar : PubMed/NCBI
3	Jennings RB, Sommers HM, Smyth GA, Flack HA and Linn H: Myocardial necrosis induced by temporary occlusion of a coronary artery in the dog. Arch Pathol. 70:68–78. 1960.PubMed/NCBI
4	Armstrong SC: Protein kinase activation and myocardial ischemia/reperfusion injury. Cardiovasc Res. 61:427–436. 2004. View Article : Google Scholar : PubMed/NCBI
5	Go AS, Mozaffarian D, Roger VL, Benjamin EJ, Berry JD, Borden WB, Bravata DM, Dai S, Ford ES, Fox CS, et al: Heart disease and stroke statistics-2013 update: A report from the American Heart Association. Circulation. 127:e6–e245. 2013. View Article : Google Scholar : PubMed/NCBI
6	Murphy E and Steenbergen C: Mechanisms underlying acute protection from cardiac ischemia-reperfusion injury. Physiol Rev. 88:581–609. 2008. View Article : Google Scholar : PubMed/NCBI
7	Becker LB: New concepts in reactive oxygen species and cardiovascular reperfusion physiology. Cardiovasc Res. 61:461–470. 2004. View Article : Google Scholar : PubMed/NCBI
8	Frangogiannis NG, Smith CW and Entman ML: The inflammatory response in myocardial infarction. Cardiovasc Res. 53:31–47. 2002. View Article : Google Scholar : PubMed/NCBI
9	Laskowski A, Woodman OL, Cao AH, Drummond GR, Marshall T, Kaye DM and Ritchie RH: Antioxidant actions contribute to the antihypertrophic effects of atrial natriuretic peptide in neonatal rat cardiomyocytes. Cardiovas Resear. 72:112–123. 2006. View Article : Google Scholar
10	Moens AL, Claeys MJ, Timmermans JP and Vrints CJ: Myocardial ischemia/reperfusion-injury, a clinical view on a complex pathophysiological process. Int J Cardiol. 100:179–190. 2005. View Article : Google Scholar : PubMed/NCBI
11	Rajak S, Banerjee SK, Sood S, Dinda AK, Gupta YK, Gupta SK and Maulik SK: Emblica officinalis causes myocardial adaptation and protects against oxidative stress in ischemic-reperfusion injury in rats. Phytother Res. 18:54–60. 2004. View Article : Google Scholar : PubMed/NCBI
12	Li J, Ichikawa T, Villacorta L, Janicki JS, Brower GL, Yamamoto M and Cui T: Nrf2 protects against maladaptive cardiac responses to hemodynamic stress. Arterioscler Thromb Vasc Biol. 29:1843–1850. 2009. View Article : Google Scholar : PubMed/NCBI
13	McMahon M, Itoh K, Yamamoto M and Hayes JD: Keap1-dependent proteasomal degradation of transcription factor Nrf2 contributes to the negative regulation of antioxidant response element-driven gene expression. J Biol Chem. 278:21592–21600. 2003. View Article : Google Scholar : PubMed/NCBI
14	Boran AD and Iyengar R: Systems approaches to polypharmacology and drug discovery. Curr Opin Drug Discov Devel. 13:297–309. 2010.PubMed/NCBI
15	Zhang A and Hui A: Inhibition of coxsackie virus and effect on the treatment of viral myocarditis of Xinji Pill in mice. Shanxi Tradit Chin Med. 12:563–564. 1998.(In Chinese).
16	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. National Academies Press (US); Washington, DC: pp. 1072–1073. 2011
17	China's food and drug administration, . Good laboratory practice. Standards Press of China; Beijing: pp. 54–59. 2003
18	China's food and drug administration: Good manufacturing practice. Standards Press of China; Beijing: pp. 106–109. 2015
19	Tao L, Gao E, Bryan NS, Qu Y, Liu HR, Hu A, Christopher TA, Lopez BL, Yodoi J, Koch WJ, et al: Cardioprotective effects of thioredoxin in myocardial ischemia and reperfusion: Role of S-nitrosation (corrected). Proc Natl Acad Sci USA. 101:11471–11476. 2004. View Article : Google Scholar : PubMed/NCBI
20	Cao Z and Li Y: Protecting against peroxynitrite-mediated cytotoxicity in vascular smooth muscle cells via upregulating endogenous glutathione biosynthesis by 3H-1,2-dithiole-3-thione. Cardiovasc Toxicol. 4:339–353. 2004. View Article : Google Scholar : PubMed/NCBI
21	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 : PubMed/NCBI
22	Hu W, Zhang P, Gu J, Yu Q and Zhang D: NEDD4-1 protects against ischaemia/reperfusion-induced cardiomyocyte apoptosis via the PI3K/Akt pathway. Apoptosis. 22:437–448. 2017. View Article : Google Scholar : PubMed/NCBI
23	Cho MH, Niles A, Huang R, Inglese J, Austin CP, Riss T and Xia M: A bioluminescent cytotoxicity assay for assessment of membrane integrity using a proteolytic biomarker. Toxicol In Vitro. 22:1099–1106. 2008. View Article : Google Scholar : PubMed/NCBI
24	Corey MJ, Kinders RJ, Brown LG and Vessella RL: A very sensitive coupled luminescent assay for cytotoxicity and complement-mediated lysis. J Immunol Methods. 207:43–51. 1997. View Article : Google Scholar : PubMed/NCBI
25	Kim H, Yoon SC, Lee TY and Jeong D: Discriminative cytotoxicity assessment based on various cellular damages. Toxicol Lett. 184:13–17. 2009. View Article : Google Scholar : PubMed/NCBI
26	Wu Y and Yuan BX: Effects of Qiangxin capsules on myocardial reperfusion arrhythmias in rats. Northwest Pharmac J. 16:23–24. 2001.
27	Fu J, Huang H, Liu J, Pi R, Chen J and Liu P: Tanshinone IIA protects cardiac myocytes against oxidative stress-triggered damage and apoptosis. Eur J Pharmac. 568:213–221. 2007. View Article : Google Scholar
28	Halliwell B and Gutteridge JMC: Free radicals in biology and medicine. 3rd. Oxford: Clarendon Press; pp. 246–350. 1999
29	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
30	Duranteau J, Chandel NS, Kulisz A, Shao Z and Schumacker PT: Intracellular signaling by reactive oxygen species during hypoxia in cardiomyocytes. J Biol Chem. 273:11619–11624. 1998. View Article : Google Scholar : PubMed/NCBI
31	Hensley K, Robinson KA, Gabbita SP, Salsman S and Floyd RA: Reactive oxygen species, cell signaling, and cell injury. Free Radic Biol Med. 28:1456–1462. 2000. View Article : Google Scholar : PubMed/NCBI
32	Das DK and Maulik N: Antioxidant effectiveness in ischemia reperfusion tissue injury. Methods Enzymol. 233:601–610. 1994. View Article : Google Scholar : PubMed/NCBI
33	Lefer DJ and Granger DN: Oxidative stress and cardiac disease. Am J Med. 109:315–323. 2000. View Article : Google Scholar : PubMed/NCBI
34	Ferrari R, Ceconi C, Curello S, Cargnoni A, Alfieri O, Pardini A, Marzollo P and Visioli O: Oxygen free radicals and myocardial damage: Protective role of thiolcontaining agents. Am J Med. 91 Suppl:95–105. 1991. View Article : Google Scholar
35	Frei B: On the role of vitamin C and other antioxidants in atherogenesis and vascular dysfunction. Proc Soc Exp Biol Med. 222:196–204. 1999. View Article : Google Scholar : PubMed/NCBI
36	Valko M, Rhodes CJ, Moncol J, Izakovic M and Mazur M: Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact. 160:1–40. 2006. View Article : Google Scholar : PubMed/NCBI
37	Tengattini S, Reiter RJ, Tan DX, Terron MP, Rodella LF and Rezzani R: Cardiovascular diseases: Protective effects of melatonin. J Pineal Res. 44:16–25. 2008.PubMed/NCBI
38	Ma Q, Kinneer K, Bi Y, Chan JY and Kan YW: Induction of murine NAD(P)H:quinine oxidoreductase by 2,3,7,8-tetrachlorodibenzo-p-dioxin requires the CNC (cap ‘n’ collar) basic leucine zipper transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2): Cross-interaction between AhR (aryl hydrocarbon receptor) and Nrf2 signal transduction. Biochem J. 377:205–213. 2004. View Article : Google Scholar : PubMed/NCBI
39	He X, Chen MG, Lin GX and Ma Q: Arsenic induces NAD(P)H-quinone oxidoreductase I by disrupting the Nrf2 × Keap1 x·Cul3 complex and recruiting Nrf2 x·Maf to the antioxidant response element enhancer. J Biol Chem. 281:23620–23631. 2006. View Article : Google Scholar : PubMed/NCBI
40	Su JD, Yen JH, Li S, Weng CY, Lin MH, Ho CT and Wu MJ: 3′,4′-didemethylnobiletin induces phase II detoxification gene expression and modulates PI3K/Akt signaling in PC12 cells. Free Radic Biol Med. 52:126–141. 2012. View Article : Google Scholar : PubMed/NCBI