Icariside II protects cardiomyocytes from hypoxia‑induced injury by upregulating the miR‑7‑5p/BTG2 axis and activating the PI3K/Akt signaling pathway

Hu,Dongxia; Gu,Yong; Wu,Dan; Zhang,Juanjuan; Li,Qing; Luo,Jun; Li,Shaochuan; Yuan,Zhen; Zhu,Bo

doi:10.3892/ijmm.2020.4677

Icariside II protects cardiomyocytes from hypoxia‑induced injury by upregulating the miR‑7‑5p/BTG2 axis and activating the PI3K/Akt signaling pathway

Authors:
- Dongxia Hu
- Yong Gu
- Dan Wu
- Juanjuan Zhang
- Qing Li
- Jun Luo
- Shaochuan Li
- Zhen Yuan
- Bo Zhu
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Affiliations: Department of Rehabilitation, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330321, P.R. China, Department of Clinical Laboratory, Jiangxi Thoracic Hospital, Nanchang, Jiangxi 330321, P.R. China, Department of Neurology, Nanchang Hongdu Hospital of TCM, Nanchang, Jiangxi 330321, P.R. China, Department of Cardiology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330321, P.R. China
Published online on: July 16, 2020 https://doi.org/10.3892/ijmm.2020.4677
Pages: 1453-1465
Copyright: © Hu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Icariside II (ICS II) has been reported to have protective effects against oxidative stress. However, whether ICS II protects cardiomyocytes from myocardial infarction (MI), and the associated underlying mechanisms, remain to be elucidated. Therefore, the current study investigated the effects of ICS II on hypoxia‑injured H9c2 cells, as well as the associated molecular mechanisms. A hypoxic injury model was established to emulate the effects of MI. The effects of ICS II on the proliferation of rat cardiomyocyte H9c2 cells were assessed with cell counting kit‑8 assays. The apoptotic status of the cells was assessed by flow cytometry, and the expression of apoptosis‑related proteins was analyzed by western blotting. A microRNA (miRNA/miR) microarray was used to quantify the differential expression of miRNAs after ICS II treatment, and the levels of miR‑7‑5p were further quantified by reverse transcription‑quantitative PCR. Whether ICS II affected hypoxia‑injured cells via miR‑7‑5p was subsequently examined, and the target of miR‑7‑5p was also investigated by bioinformatics analysis and luciferase reporter assays. The effects of ICS II on the PI3K/Akt pathway were then evaluated by western blot analysis. Hypoxia treatment decreased viability and the migration and invasion abilities of H9c2 cells, and also induced apoptosis. ICS II significantly increased viability and reduced hypoxia‑associated apoptosis. Moreover, ICS II treatment led to the upregulation of miR‑7‑5p, and the protective effects of ICS II were found to rely on miR‑7‑5p. Moreover, BTG anti‑proliferation factor (BTG2) was identified as a direct target of miR‑7‑5p, and overexpression of BTG2 inhibited the protective effects of miR‑7‑5p. Finally, ICS II treatment resulted in the activation of the PI3K/Akt signaling pathway, which is essential for the survival of H9c2 cells under hypoxic conditions. In summary, ICS II reduces hypoxic injury in H9c2 cells via the miR‑7‑5p/BTG2 axis and activation of the PI3K/Akt signaling pathway.

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1	Townsend N, Wilson L, Bhatnagar P, Wickramasinghe K, Rayner M and Nichols M: Cardiovascular disease in Europe Epidemiological update 2016. Eur Heart J. 37:3232–3245. 2016. View Article : Google Scholar : PubMed/NCBI
2	Yellon DM and Hausenloy DJ: Myocardial reperfusion injury. N Engl J Med. 357:1121–1135. 2007. View Article : Google Scholar : PubMed/NCBI
3	Graham RM, Frazier DP, Thompson JW, Haliko S, Li H, Wasserlauf BJ, Spiga MG, Bishopric NH and Webster KA: A unique pathway of cardiac myocyte death caused by hypoxia-acidosis. J Exp Biol. 207:3189–3200. 2004. View Article : Google Scholar : PubMed/NCBI
4	Kemp CD and Conte JV: The pathophysiology of heart failure. Cardiovasc Pathol. 21:365–371. 2012. View Article : Google Scholar : PubMed/NCBI
5	Ambrosy AP, Fonarow GC, Butler J, Chioncel O, Greene SJ, Vaduganathan M, Nodari S, Lam CSP, Sato N, Shah AN and Gheorghiade M: The global health and economic burden of hospitalizations for heart failure: Lessons learned from hospitalized heart failure registries. J Am Coll Cardiol. 63:1123–1133. 2014. View Article : Google Scholar : PubMed/NCBI
6	Xie F, Wu CF, Lai WP, Yang XJ, Cheung PY, Yao XS, Leung PC and Wong MS: The osteoprotective effect of Herba epimedii (HEP) extract in vivo and in vitro. Evid Based Complement Alternat Med. 2:353–361. 2005. View Article : Google Scholar : PubMed/NCBI
7	Chen M, Wu J, Luo Q, Mo S, Lyu Y, Wei Y and Dong J: The anticancer properties of Herba epimedii and its main bioactive componentsicariin and icariside II. Nutrients. 8:E5632016. View Article : Google Scholar : PubMed/NCBI
8	Fu S, Li YL, Wu YT, Yue Y, Qian ZQ and Yang DL: Icariside II attenuates myocardial fibrosis by inhibiting nuclear factor-κB and the TGF-β1/Smad2 signalling pathway in spontaneously hypertensive rats. Biomed Pharmacother. 100:64–71. 2018. View Article : Google Scholar : PubMed/NCBI
9	Yang L, Peng C, Xia J, Zhang W, Tian L, Tian Y, Yang X and Cao Y: Effects of icariside II ameliorates diabetic cardiomyopathy in streptozotocin-induced diabetic rats by activating Akt/NOS/NF-κB signaling. Mol Med Rep. 17:4099–4105. 2018.
10	Alvarez-Garcia I and Miska EA: MicroRNA functions in animal development and human disease. Development. 132:4653–4662. 2005. View Article : Google Scholar : PubMed/NCBI
11	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
12	Boon RA and Dimmeler S: MicroRNAs in myocardial infarction. Nat Rev Cardiol. 12:135–142. 2015. View Article : Google Scholar
13	Gu Y, Liang Z, Wang H, Jin J, Zhang S, Xue S, Chen J, He H, Duan K, Wang J, et al: Tanshinone IIA protects H9c2 cells from oxidative stress-induced cell death via microRNA-133 upregulation and Akt activation. Exp Ther Med. 12:1147–1152. 2016. View Article : Google Scholar : PubMed/NCBI
14	Jiang G, Wu H, Hu Y, Li J and Li Q: Gastrodin inhibits glutamate-induced apoptosis of PC12 cells via inhibition of CaMKII/ASK-1/p38 MAPK/p53 signaling cascade. Cell Mol Neurobiol. 34:591–602. 2014. View Article : Google Scholar : PubMed/NCBI
15	Kalinowski FC, Brown RA, Ganda C, Giles KM, Epis MR, Horsham J and Leedman PJ: microRNA-7: A tumor suppressor miRNA with therapeutic potential. Int J Biochem Cell Biol. 54:312–317. 2014. View Article : Google Scholar : PubMed/NCBI
16	Li B, Li R, Zhang C, Bian HJ, Wang F, Xiao J, Liu SW, Yi W, Zhang MX, Wang SX, et al: MicroRNA-7a/b protects against cardiac myocyte injury in ischemia/reperfusion by targeting poly(ADP-ribose) polymerase. PLoS One. 9:e900962014. View Article : Google Scholar : PubMed/NCBI
17	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
18	Song J, Feng L, Zhong R, Xia Z, Zhang L, Cui L, Yan H, Jia X and Zhang Z: Icariside II inhibits the EMT of NSCLC cells in inflammatory microenvironment via down-regulation of Akt/NF-κB signaling pathway. Mol Carcinog. 56:36–48. 2017. View Article : Google Scholar
19	Khan M, Maryam A, Qazi JI and Ma T: Targeting apoptosis and multiple signaling pathways with icariside II in cancer cells. Int J Biol Sci. 11:1100–1112. 2015. View Article : Google Scholar : PubMed/NCBI
20	Gao J, Deng Y, Yin C, Liu Y, Zhang W, Shi J and Gong Q: Icariside II, a novel phosphodiesterase 5 inhibitor, protects against H₂ O₂ -induced PC12 cells death by inhibiting mitochondria-mediated autophagy. J Cell Mol Med. 21:375–386. 2017. View Article : Google Scholar
21	Li J, Qiu M, An Y, Huang J and Gong C: miR-7-5p acts as a tumor suppressor in bladder cancer by regulating the hedgehog pathway factor Gli3. Biochem Biophys Res Commun. 503:2101–2107. 2018. View Article : Google Scholar : PubMed/NCBI
22	Zhu W, Wang Y, Zhang D, Yu X and Leng X: MiR-7-5p functions as a tumor suppressor by targeting SOX18 in pancreatic ductal adenocarcinoma. Biochem Biophys Res Commun. 497:963–970. 2018. View Article : Google Scholar : PubMed/NCBI
23	Giles KM, Brown RA, Ganda C, Podgorny MJ, Candy PA, Wintle LC, Richardson KL, Kalinowski FC, Stuart LM, Epis MR, et al: microRNA-7-5p inhibits melanoma cell proliferation and metastasis by suppressing RelA/NF-κB. Oncotarget. 7:31663–31680. 2016. View Article : Google Scholar : PubMed/NCBI
24	Zou L, Ma X, Lin S, Wu B, Chen Y and Peng C: Long noncoding RNA-MEG3 contributes to myocardial ischemia-reperfusion injury through suppression of miR-7-5p expression. Biosci Rep. 39:BSR201902102019. View Article : Google Scholar : PubMed/NCBI
25	Buanne P, Corrente G, Micheli L, Palena A, Lavia P, Spadafora C, Lakshmana MK, Rinaldi A, Banfi S, Quarto M, et al: Cloning of PC3B, a novel member of the PC3/BTG/TOB family of growth inhibitory genes, highly expressed in the olfactory epithelium. Genomics. 68:253–263. 2000. View Article : Google Scholar : PubMed/NCBI
26	Yuniati L, Scheijen B, van der Meer LT and van Leeuwen FN: Tumor suppressors BTG1 and BTG2: Beyond growth control. J Cell Physiol. 234:5379–5389. 2019. View Article : Google Scholar :
27	Tong Z, Jiang B, Wu Y, Liu Y, Li Y, Gao M, Jiang Y, Lv Q and Xiao X: MiR-21 protected cardiomyocytes against doxorubicin-induced apoptosis by targeting BTG2. Int J Mol Sci. 16:14511–14525. 2015. View Article : Google Scholar : PubMed/NCBI
28	Leszczynska KB, Foskolou IP, Abraham AG, Anbalagan S, Tellier C, Haider S, Span PN, O'Neill EE, Buffa FM and Hammond EM: Hypoxia-induced p53 modulates both apoptosis and radiosensitivity via AKT. J Clin Invest. 125:2385–2398. 2015. View Article : Google Scholar : PubMed/NCBI
29	Bell RM and Yellon DM: Bradykinin limits infarction when administered as an adjunct to reperfusion in mouse heart: The role of PI3K, Akt and eNOS. J Mol Cell Cardiol. 35:185–193. 2003. View Article : Google Scholar : PubMed/NCBI
30	Keyes KT, Xu J, Long B, Zhang C, Hu Z and Ye Y: Pharmacological inhibition of PTEN limits myocardial infarct size and improves left ventricular function postinfarction. Am J Physiol Heart Circ Physiol. 298:H1198–H1208. 2010. View Article : Google Scholar : PubMed/NCBI
31	Liu W, Mao L, Ji F, Chen F, Wang S and Xie Y: Icariside II activates EGFR-Akt-Nrf2 signaling and protects osteoblasts from dexamethasone. Oncotarget. 8:2594–2603. 2017. View Article : Google Scholar :
32	Liu ML, Zhang Q, Yuan X, Jin L, Wang LL, Fang TT and Wang WB: Long noncoding RNA RP4 functions as a competing endogenous RNA through miR-7-5p sponge activity in colorectal cancer. World J Gastroenterol. 24:1004–1012. 2018. View Article : Google Scholar : PubMed/NCBI
33	Xu Z, Wang Y, Xiong J, Cui F, Wang L and Peng H: NUSAP1 knockdown inhibits cell growth and metastasis of non-small-cell lung cancer through regulating BTG2/PI3K/Akt signaling. J Cell Physiol. 235:3886–3893. 2019. View Article : Google Scholar : PubMed/NCBI