miR-181b-5p suppresses starvation-induced cardiomyocyte autophagy by targeting Hspa5

Chang,Liuhui; Chai,Xiaoming; Chen,Peiming; Cao,Jianfang; Xie,Hong; Zhu,Jiang

doi:10.3892/ijmm.2018.3988

miR-181b-5p suppresses starvation-induced cardiomyocyte autophagy by targeting Hspa5

Authors:
- Liuhui Chang
- Xiaoming Chai
- Peiming Chen
- Jianfang Cao
- Hong Xie
- Jiang Zhu
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Affiliations: Department of Anesthesiology, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China
Published online on: November 12, 2018 https://doi.org/10.3892/ijmm.2018.3988
Pages: 143-154
Copyright: © Chang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

This study aimed to investigate the role of microRNA‑181b‑5p (miR‑181b‑5p) in starvation‑induced cardiomyocyte autophagy by targeting heat shock protein family A member 5 (Hspa5). For this purpose, H9c2 cardiomyocytes and neonatal rat ventricular myocytes (NRVMs) were glucose‑starved in Earle's Balanced Salt Solution (EBSS) for different periods of time (0, 2, 4, 6 and 8 h). RT‑qPCR analysis was performed to examine the expression of miR‑181b‑5p in the different groups. Immunofluorescence was performed to detect the expression of LC3. In addition, the H9c2 cardiomyocytes and NRVMs were transfected with miR‑181b‑5p mimic, miR‑181b‑5p inhibitor, siHspa5 or their respective controls. An MTT assay was performed to measure cell proliferation in the different groups. Western blot analysis was performed to determine the expression of Beclin‑1, Hspa5, phosphorylated phosphoinositide 3‑kinase PI3K (p‑PI3K), phosphorylated Akt (p‑Akt), phosphorylated mammalian target of rapamycin (p‑mTOR), Bcl‑2, Bax and cleaved caspase‑3. Flow cytometry was performed to assess cell apoptosis. A luciferase reporter assay was performed to determine whether Hspa5 is a direct target of miR‑181b‑5p. The results revealed that the downregulation of miR‑181b‑5p promoted cell autophagy in the cardiomyocytes. Moreover, miR‑181b‑5p negatively regulated Beclin‑1 and Hspa5. Beclin‑1 is a well‑known autophagy‑ and apoptosis‑related protein. In addition, cell apoptosis was attenuated by the decreased expression of miR‑181b‑5p in the cardiomyocytes. Bcl‑2 prevented apoptosis and autophagy by binding to Bax and Bcl‑2, respectively. The upregulation of miR‑181b‑5p inhibited autophagy and promoted apoptosis via Hspa5. miR‑181b‑5p inhibition promoted p‑mTOR, p‑Akt and p‑PI3K expression via Hspa5. The results of luciferase reporter assay also confirmed that Hspa5 is a direct target of miR‑181b‑5p. On the whole, the findings of this study suggest that miR‑181b‑5p contributes to starvation‑induced autophagy and apoptosis in cardiomyocytes by directly targeting Hspa5 via the PI3K/Akt/mTOR signaling pathway.

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1	Benjamin EJ, Virani SS, Callaway CW, Chamberlain AM, Chang AR, Cheng S, Chiuve SE, Cushman M, Delling FN, Deo R, et al: Heart disease and stroke statistics-2018 update: A report from the american heart Association. Circulation. 137:e67–e492. 2018. View Article : Google Scholar : PubMed/NCBI
2	Shimomura H, Terasaki F, Hayashi T, Kitaura Y, Isomura T and Suma H: Autophagic degeneration as a possible mechanism of myocardial cell death in dilated cardiomyopathy. Jpn Circ J. 65:965–968. 2001. View Article : Google Scholar : PubMed/NCBI
3	Ma S, Wang Y, Chen Y and Cao F: The role of the autophagy in myocardial ischemia/reperfusion injury. Biochim Biophys Acta. 1852.271–276. 2015.
4	Wang K, Liu CY, Zhou LY, Wang JX, Wang M, Zhao B, Zhao WK, Xu SJ, Fan LH, Zhang XJ, et al: APF lncRNA regulates autophagy and myocardial infarction by targeting miR-188-3p. Nat Commun. 6:67792015. View Article : Google Scholar : PubMed/NCBI
5	Wu H, Wang Y, Wang X, Li R and Yin D: MicroRNA-365 accelerates cardiac hypertrophy by inhibiting autophagy via the modulation of Skp2 expression. Biochem Biophys Res Commun. 484:304–310. 2017. View Article : Google Scholar : PubMed/NCBI
6	Takemura G, Kanamori H, Okada H, Miyazaki N, Watanabe T, Tsujimoto A, Goto K, Maruyama R, Fujiwara T and Fujiwara H: Anti-apoptosis in nonmyocytes and pro-autophagy in cardiomyocytes: Two strategies against postinfarction heart failure through regulation of cell death/degeneration. Heart Fail Rev. 23:759–772. 2018. View Article : Google Scholar : PubMed/NCBI
7	Palikaras K, Lionaki E and Tavernarakis N: Mitophagy: In sickness and in health. Mol Cell Oncol. 3:e10563322015. View Article : Google Scholar
8	Ortiz C and Cardemil L: Heat-shock responses in two leguminous plants: A comparative study. J Exp Bot. 52:1711–1719. 2001.PubMed/NCBI
9	Petrovski G, Das S, Juhasz B, Kertesz A, Tosaki A and Das DK: Cardioprotection by endoplasmic reticulum stress-induced autophagy. Antioxid Redox Signal. 14:2191–2200. 2011. View Article : Google Scholar
10	Eizirik DL, Cardozo AK and Cnop M: The role for endoplasmic reticulum stress in diabetes mellitus. Endocr Rev. 29:42–61. 2008. View Article : Google Scholar
11	Yorimitsu T, Nair U, Yang Z and Klionsky DJ: Endoplasmic reticulum stress triggers autophagy. J Biol Chem. 281:30299–30304. 2006. View Article : Google Scholar : PubMed/NCBI
12	Wang L, Hong Q, Lv Y, Feng Z, Zhang X, Wu L, Cui S, Hou K, Su H, Huang Z, et al: Autophagy can repair endoplasmic reticulum stress damage of the passive Heymann nephritis model as revealed by proteomics analysis. J Proteomics. 75:3866–3876. 2012. View Article : Google Scholar : PubMed/NCBI
13	Zhang PL, Lun M, Teng J, Huang J, Blasick TM, Yin L, Herrera GA and Cheung JY: Preinduced molecular chaperones in the endoplasmic reticulum protect cardiomyocytes from lethal injury. Ann Clin Lab Sci. 34:449–457. 2004.
14	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
15	Zhang Z, Li H, Chen S, Li Y, Cui Z and Ma J: Knockdown of MicroRNA-122 protects H9c2 cardiomyocytes from hypoxia-induced apoptosis and promotes autophagy. Med Sci Monit. 23:4284–4290. 2017. View Article : Google Scholar : PubMed/NCBI
16	Zhi F, Wang Q, Deng D, Shao N, Wang R, Xue L, Wang S, Xia X and Yang Y: MiR-181b-5p downregulates NOVA1 to suppress proliferation, migration and invasion and promote apoptosis in astrocytoma. PLoS One. 9:e1091242014. View Article : Google Scholar : PubMed/NCBI
17	Huo X, Zhang K, Yi L, Mo Y, Liang Y, Zhao J, Zhang Z, Xu Y and Zhen G: Decreased epithelial and plasma miR-181b-5p expression associates with airway eosinophilic inflammation in asthma. Clin Exp Allergy. 46:1281–1290. 2016. View Article : Google Scholar : PubMed/NCBI
18	Alacam H, Akgun S, Akca H, Ozturk O, Kabukcu BB and Herken H: miR-181b-5p miR-195-5p and miR-301a-3p are related with treatment resistance in schizophrenia. Psychiatry Res. 245:200–206. 2016. View Article : Google Scholar : PubMed/NCBI
19	Lin R, Su Z, Tan X, Su Y, Chen Y, Shu X, Liang S, Wang J and Xie S: Effect of endoplasmic reticulum stress and autophagy in the regulation of post-infarct cardiac repair. Arch Med Res. 2018. View Article : Google Scholar
20	Yang Z and Klionsky DJ: An overview of the molecular mechanism of autophagy. Curr Top Microbiol Immunol. 335:1–32. 2009.PubMed/NCBI
21	He C and Klionsky DJ: Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet. 43:67–93. 2009. View Article : Google Scholar : PubMed/NCBI
22	Tomihara H, Yamada D, Eguchi H, Iwagami Y, Noda T, Asaoka T, Wada H, Kawamoto K, Gotoh K, Takeda Y, et al: MicroRNA-181b-5p ETS1, and the c-Met pathway exacerbate the prognosis of pancreatic ductal adenocarcinoma after radiation therapy. Cancer Sci. 108:398–407. 2017. View Article : Google Scholar : PubMed/NCBI
23	Tian F, Shen Y, Chen Z, Li R, Lu J and Ge Q: Aberrant miR-181b-5p and miR-486-5p expression in serum and tissue of non-small cell lung cancer. Gene. 591:338–343. 2016. View Article : Google Scholar : PubMed/NCBI
24	Copier CU, Leon L, Fernandez M, Contador D and Calligaris SD: Circulating miR-19b and miR-181b are potential biomarkers for diabetic cardiomyopathy. Sci Rep. 7:135142017. View Article : Google Scholar : PubMed/NCBI
25	Marques FZ, Vizi D, Khammy O, Mariani JA and Kaye DM: The transcardiac gradient of cardio-microRNAs in the failing heart. Eur J Heart Fail. 18:1000–1008. 2016. View Article : Google Scholar : PubMed/NCBI
26	Zhong Y, Wang QJ, Li X, Yan Y, Backer JM, Chait BT, Heintz N and Yue Z: Distinct regulation of autophagic activity by Atg14L and Rubicon associated with Beclin 1- phosphatidylinositol-3-kin ase complex. Nat Cell Biol. 11:468–476. 2009. View Article : Google Scholar : PubMed/NCBI
27	Hamacher-Brady A, Brady NR and Gottlieb RA: Enhancing macroautophagy protects against ischemia/reperfusion injury in cardiac myocytes. J Biol Chem. 281:29776–29787. 2006. View Article : Google Scholar : PubMed/NCBI
28	Lin M, Tang S, Zhang C, Chen H, Huang W, Liu Y and Zhang J: Euphorbia factor L2 induces apoptosis in A549 cells through the mitochondrial pathway. Acta Pharm Sin B. 7:59–64. 2017. View Article : Google Scholar : PubMed/NCBI
29	Tao YW, Lin YC, She ZG, Lin MT, Chen PX and Zhang JY: Anticancer activity and mechanism investigation of beauvericin isolated from secondary metabolites of the mangrove endophytic fungi. Anticancer Agents Med Chem. 15:258–266. 2015. View Article : Google Scholar : PubMed/NCBI
30	Zhang JY, Lin MT, Tung HY, Tang SL, Yi T, Zhang YZ, Tang YN, Zhao ZZ and Chen HB: Bruceine D induces apoptosis in human chronic myeloid leukemia K562 cells via mitochondrial pathway. Am J Cancer Res. 6:819–826. 2016.PubMed/NCBI
31	Chen L, Wang FY, Zeng ZY, Cui L, Shen J, Song XW, Li P, Zhao XX and Qin YW: MicroRNA-199a acts as a potential suppressor of cardiomyocyte autophagy through targeting Hspa5. Oncotarget. 8:63825–63834. 2017.PubMed/NCBI
32	Hou X, Hu Z, Xu H, Xu J, Zhang S, Zhong Y, He X and Wang N: Advanced glycation endproducts trigger autophagy in cadiomyocyte via RAGE/PI3K/AKT/mTOR pathway. Cardiovasc Diabetol. 13:782014. View Article : Google Scholar : PubMed/NCBI
33	Yu W, Sun H, Zha W, Cui W, Xu L, Min Q and Wu J: Apigenin attenuates adriamycin-Induced cardiomyocyte apoptosis via the PI3K/AKT/mTOR pathway. Evid Based Complement Alternat Med. 2017.2590676:2017.