Open Access

Tetramethylpyrazine protects retinal ganglion cells against H2O2‑induced damage via the microRNA‑182/mitochondrial pathway

  • Authors:
    • Xinmin Li
    • Qiuli Wang
    • Yanfan Ren
    • Xiaomin Wang
    • Huaxu Cheng
    • Hua Yang
    • Baojun Wang
  • View Affiliations

  • Published online on: May 29, 2019     https://doi.org/10.3892/ijmm.2019.4214
  • Pages: 503-512
  • Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Glaucoma is the leading cause of irreversible blindness worldwide; the apoptosis of the retinal ganglion cells (RGCs) is a hallmark of glaucoma. Tetramethylpyrazine (TMP) is the main active component of Ligusticum wallichii Franchat, and has been demonstrated to improve a variety of injuries through its antioxidative and antiapoptotic properties. However, these effects of TMP on glaucoma have not been studied. The present study aimed to investigate the potential role of TMP in glaucoma and to elucidate its possible mechanisms responsible for these effects. An in vitro model was generated, in which primary RGCs (PRGCs) were treated with H2O2. Our study revealed that TMP protected against H2O2‑induced injury to PRGCs, as evidenced by enhanced cell viability, reduced caspase 3 activity and decreased cell apoptosis. We also reported that TMP treatment inhibited reactive oxygen species (ROS) production and malondialdehyde levels, but upregulated the antioxidative enzyme superoxide dismutase. In particular, TMP significantly increased the expression of microRNA‑182‑5p (miR‑182) in H2O2‑treated PRGCs, which was selected as the target miRNA for further research. In addition, our findings suggested that the protective effects of TMP on H2O2‑induced injury were attenuated by knockdown of miR‑182. The results of a luciferase reporter assay demonstrated that Bcl‑2 interacting protein 3 (BNIP3), an effector of mitochondria‑mediated apoptosis, was a direct target of miR‑182. In addition, TMP treatment significantly decreased the expression of BNIP3, Bax, cleaved‑caspase‑3 and cleaved‑poly(ADP‑ribose)polymerase, but increased that of Bcl‑2. Also, TMP treatment decreased the release of cytochrome c from mitochondria and improved mitochondrial membrane potential in H2O2‑treated RGCs. Of note, the inhibitory effects of TMP on the mitochondrial apoptotic pathway were suggested to be reversed by knockdown of miR‑182. Collectively, our findings provide novel evidence that TMP protects PRGCs against H2O2‑induced damage through suppressing apoptosis and oxidative stress via the miR‑182/mitochondrial apoptotic pathway.

References

1 

Koriyama Y, Ohno M, Kimura T and Kato S: Neuroprotective effects of 5-S-GAD against oxidative stress-induced apoptosis in RGC-5 cells. Brain Res. 1296:187–195. 2009. View Article : Google Scholar : PubMed/NCBI

2 

Tham YC, Li X, Wong TY, Quigley HA, Aung T and Cheng CY: Global prevalence of glaucoma and projections of glaucoma burden through 2040: A systematic review and meta-analysis. Ophthalmology. 121:2081–2090. 2014. View Article : Google Scholar : PubMed/NCBI

3 

Lee D, Shim MS, Kim KY, Noh YH, Kim H, Kim SY, Weinreb RN and Ju WK: Coenzyme Q10 inhibits glutamate excitotoxicity and oxidative stress-mediated mitochondrial alteration in a mouse model of glaucoma. Invest Ophthalmol Vis Sci. 55:993–1005. 2014. View Article : Google Scholar : PubMed/NCBI

4 

Sancho P, Fernández C, Yuste VJ, Amrán D, Ramos AM, de Blas E, Susin SA and Aller P: Regulation of apoptosis/necrosis execution in cadmium-treated human promonocytic cells under different forms of oxidative stress. Apoptosis. 11:673–686. 2006. View Article : Google Scholar : PubMed/NCBI

5 

Almasieh M, Wilson AM, Morquette B, Cueva Vargas JL and Di Polo A: The molecular basis of retinal ganglion cell death in glaucoma. Prog Retin Eye Res. 31:152–181. 2012. View Article : Google Scholar

6 

Zhai L, Zhang P, Sun RY, Liu XY, Liu WG and Guo XL: Cytoprotective effects of CSTMP, a novel stilbene derivative, against H2O2-induced oxidative stress in human endothelial cells. Pharmacol Rep. 63:1469–1480. 2011. View Article : Google Scholar

7 

Wu J, Song R, Song W, Li Y, Zhang Q, Chen Y, Fu Y, Fang W, Wang J, Zhong Z, et al: Chlorpromazine protects against apoptosis induced by exogenous stimuli in the developing rat brain. PLoS One. 6:e219662011. View Article : Google Scholar : PubMed/NCBI

8 

Tang Z, Wang Q, Xu H and Zhang W: Microdialysis sampling for investigations of tetramethylpyrazine following transdermal and intraperitoneal administration. Eur J Pharm Sci. 50:454–458. 2013. View Article : Google Scholar : PubMed/NCBI

9 

Gong X, Wang Q, Tang X, Wang Y, Fu D, Lu H, Wang G and Norgren S: Tetramethylpyrazine prevents contrast-induced nephropathy by inhibiting p38 MAPK and FoxO1 signaling pathways. Am J Nephrol. 37:199–207. 2013. View Article : Google Scholar : PubMed/NCBI

10 

Yang G, Qian C, Wang N, Lin C, Wang Y, Wang G and Piao X: Tetramethylpyrazine protects against oxygen-glucose deprivation-induced brain microvascular endothelial cells injury via Rho/Rho-kinase signaling pathway. Cell Mol Neurobiol. 37:619–633. 2017. View Article : Google Scholar

11 

Lu C, Zhang J, Shi X, Miao S, Bi L, Zhang S, Yang Q, Zhou X, Zhang M, Xie Y, et al: Neuroprotective effects of tetramethyl-pyrazine against dopaminergic neuron injury in a rat model of Parkinson's disease induced by MPTP. Int J Biol Sci. 10:350–357. 2014. View Article : Google Scholar :

12 

Zhong M, Ma W, Zhang X, Wang Y and Gao X: Tetramethyl pyrazine protects hippocampal neurons against anoxia/reoxygenation injury through inhibiting apoptosis mediated by JNK/MARK signal pathway. Med Sci Monit. 22:5082–5090. 2016. View Article : Google Scholar : PubMed/NCBI

13 

Luo X, Yu Y, Xiang Z, Wu H, Ramakrishna S, Wang Y, So KF, Zhang Z and Xu Y: Tetramethylpyrazine nitrone protects retinal ganglion cells against N-methyl-d-aspartate-induced excitotoxicity. J Neurochem. 141:373–386. 2017. View Article : Google Scholar : PubMed/NCBI

14 

Ambros V: The functions of animal microRNAs. Nature. 431:350–355. 2004. View Article : Google Scholar : PubMed/NCBI

15 

Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI

16 

Guo R, Shen W, Su C, Jiang S and Wang J: Relationship between the Pathogenesis of Glaucoma and miRNA. Ophthalmic Res. 57:194–199. 2017. View Article : Google Scholar : PubMed/NCBI

17 

Kong N, Lu X and Li B: Downregulation of microRNA-100 protects apoptosis and promotes neuronal growth in retinal ganglion cells. BMC Mol Biol. 15:252014. View Article : Google Scholar : PubMed/NCBI

18 

Li H, Zhu Z, Liu J, Wang J and Qu C: MicroRNA-137 regulates hypoxia-induced retinal ganglion cell apoptosis through Notch1. Int J Mol Med. 41:1774–1782. 2018.

19 

Zhang QL, Wang W, Alatantuya, Dongmei, Lu ZJ, Li LL and Zhang TZ: Down-regulated miR-187 promotes oxidative stress-induced retinal cell apoptosis through P2X7 receptor. Int J Biol Macromol. 120:801–810. 2018. View Article : Google Scholar : PubMed/NCBI

20 

Cheng LB, Li KR, Yi N, Li XM, Wang F, Xue B, Pan YS, Yao J, Jiang Q and Wu ZF: miRNA-141 attenuates UV-induced oxida-tive stress via activating Keap1-Nrf2 signaling in human retinal pigment epithelium cells and retinal ganglion cells. Oncotarget. 8:13186–13194. 2017.PubMed/NCBI

21 

Jiao J, Huang X, Feit-Leithman RA, Neve RL, Snider W, Dartt DA and Chen DF: Bcl-2 enhances Ca(2+) signaling to support the intrinsic regenerative capacity of CNS axons. EMBO J. 24:1068–1078. 2005. View Article : Google Scholar : PubMed/NCBI

22 

Rodriguez AR, de Sevilla Müller LP and Brecha NC: The RNA binding protein RBPMS is a selective marker of ganglion cells in the mammalian retina. J Comp Neurol. 522:1411–1443. 2014. View Article : Google Scholar :

23 

Zhang XM, Li Liu DT, Chiang SW, Choy KW, Pang CP, Lam DS and Yam GH: Immunopanning purification and long-term culture of human retinal ganglion cells. Mol Vis. 16:2867–2872. 2010.

24 

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

25 

Lee SY, Lee S, Choi E, Ham O, Lee CY, Lee J, Seo HH, Cha MJ, Mun B, Lee Y, et al: Small molecule-mediated up-regulation of microRNA targeting a key cell death modulator BNIP3 improves cardiac function following ischemic injury. Sci Rep. 6:234722016. View Article : Google Scholar : PubMed/NCBI

26 

Ju WK, Liu Q, Kim KY, Crowston JG, Lindsey JD, Agarwal N, Ellisman MH, Perkins GA and Weinreb RN: Elevated hydrostatic pressure triggers mitochondrial fission and decreases cellular ATP in differentiated RGC-5 cells. Invest Ophthalmol Vis Sci. 48:2145–2151. 2007. View Article : Google Scholar : PubMed/NCBI

27 

Liu Q, Ju WK, Crowston JG, Xie F, Perry G, Smith MA, Lindsey JD and Weinreb RN: Oxidative stress is an early event in hydrostatic pressure induced retinal ganglion cell damage. Invest Ophthalmol Vis Sci. 48:4580–4589. 2007. View Article : Google Scholar : PubMed/NCBI

28 

Lv B, Chen T, Xu Z, Huo F, Wei Y and Yang X: Crocin protects retinal ganglion cells against H2O2-induced damage through the mitochondrial pathway and activation of NF-κB. Int J Mol Med. 37:225–232. 2016. View Article : Google Scholar : PubMed/NCBI

29 

Zhang QL, Wang W, Jiang Y, A-Tuya, Dongmei, Li LL, Lu ZJ, Chang H and Zhang TZ: GRGM-13 comprising 13 plant and animal products, inhibited oxidative stress induced apoptosis in retinal ganglion cells by inhibiting P2RX7/p38 MAPK signaling pathway. Biomed Pharmacother. 101:494–500. 2018. View Article : Google Scholar : PubMed/NCBI

30 

Chen H, Chow PH, Cheng SK, Cheung AL, Cheng LY and O WS: Male genital tract antioxidant enzymes: Their source, function in the female, and ability to preserve sperm DNA integrity in the golden hamster. J Androl. 24:704–711. 2003. View Article : Google Scholar : PubMed/NCBI

31 

Lin J, Chuang CC and Zuo L: Potential roles of microRNAs and ROS in colorectal cancer: Diagnostic biomarkers and therapeutic targets. Oncotarget. 8:17328–17346. 2017.PubMed/NCBI

32 

Lan J, Huang Z, Han J, Shao J and Huang C: Redox regulation of microRNAs in cancer. Cancer Lett. 418:250–259. 2018. View Article : Google Scholar : PubMed/NCBI

33 

Liu Y, Qiang W, Xu X, Dong R, Karst AM, Liu Z, Kong B, Drapkin RI and Wei JJ: Role of miR-182 in response to oxidative stress in the cell fate of human fallopian tube epithelial cells. Oncotarget. 6:38983–38998. 2015.PubMed/NCBI

34 

Lv G, Shao S, Dong H, Bian X, Yang X and Dong S: MicroRNA-214 protects cardiac myocytes against H2O2-induced injury. J Cell Biochem. 115:93–101. 2014. View Article : Google Scholar

35 

Qin SB, Peng DY, Lu JM and Ke ZP: MiR-182-5p inhibited oxidative stress and apoptosis triggered by oxidized low-density lipoprotein via targeting toll-like receptor 4. J Cell Physiol. 233:6630–6637. 2018. View Article : Google Scholar

36 

Li J and Li J, Wei T and Li J: Down-regulation of MicroRNA-137 improves high glucose-induced oxidative stress injury in human umbilical vein endothelial cells by up-regulation of AMPKα1. Cell Physiol Biochem. 39:847–859. 2016. View Article : Google Scholar

37 

Shi YF, Liu N, Li YX, Song CL, Song XJ, Zhao Z and Liu B: Insulin protects H9c2 rat cardiomyoblast cells against hydrogen peroxide-induced injury through upregulation of microRNA-210. Free Radic Res. 49:1147–1155. 2015. View Article : Google Scholar : PubMed/NCBI

38 

Li X, Kong M, Jiang D, Qian J, Duan Q and Dong A: MicroRNA-150 aggravates H2O2-induced cardiac myocyte injury by down-regulating c-myb gene. Acta Biochim Biophys Sin (Shanghai). 45:734–741. 2013. View Article : Google Scholar

39 

Li QC, Xu H, Wang X, Wang T and Wu J: miR-34a increases cisplatin sensitivity of osteosarcoma cells in vitro through up-regulation of c-Myc and Bim signal. Cancer Biomark. 21:135–144. 2017. View Article : Google Scholar : PubMed/NCBI

40 

Lomonosova E and Chinnadurai G: BH3-only proteins in apop-tosis and beyond: An overview. Oncogene. 27(Suppl 1): S2–S19. 2008. View Article : Google Scholar

41 

Quinsay MN, Lee Y, Rikka S, Sayen MR, Molkentin JD, Gottlieb RA and Gustafsson AB: Bnip3 mediates permeabilization of mitochondria and release of cytochrome c via a novel mechanism. J Mol Cell Cardiol. 48:1146–1156. 2010. View Article : Google Scholar :

42 

Circu ML and Aw TY: Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic Biol Med. 48:749–762. 2010. View Article : Google Scholar : PubMed/NCBI

43 

Cai X, Chen Z, Pan X, Xia L, Chen P, Yang Y, Hu H, Zhang J, Li K, Ge J, et al: Inhibition of angiogenesis, fibrosis and thrombosis by tetramethylpyrazine: Mechanisms contributing to the SDF-1/CXCR4 axis. PLoS One. 9:e881762014. View Article : Google Scholar : PubMed/NCBI

44 

Yang Z, Zhang Q, Ge J and Tan Z: Protective effects of tetra-methylpyrazine on rat retinal cell cultures. Neurochem Int. 52:1176–1187. 2008. View Article : Google Scholar : PubMed/NCBI

45 

Fu YS, Lin YY, Chou SC, Tsai TH, Kao LS, Hsu SY, Cheng FC, Shih YH, Cheng H, Fu YY and Wang JY: Tetramethylpyrazine inhibits activities of glioma cells and glutamate neuro-excitotox-icity: Potential therapeutic application for treatment of gliomas. Neuro Oncol. 10:139–152. 2008. View Article : Google Scholar : PubMed/NCBI

46 

Wang K, Zhu X, Zhang K, Zhou F and Zhu L: Neuroprotective effect of tetramethylpyrazine against all-trans-retinal toxicity in the differentiated Y-79 cells via upregulation of IRBP expression. Exp Cell Res. 359:120–128. 2017. View Article : Google Scholar : PubMed/NCBI

47 

Yu N, Zhang Z, Chen P, Zhong Y, Cai X, Hu H, Yang Y, Zhang J, Li K, Ge J, et al: Tetramethylpyrazine (TMP), an active ingredient of chinese herb medicine chuanxiong, attenuates the degeneration of trabecular meshwork through SDF-1/CXCR4 axis. PLoS One. 10:e01330552015. View Article : Google Scholar : PubMed/NCBI

48 

Izzotti A, Sacca SC, Cartiglia C and De Flora S: Oxidative deoxyribonucleic acid damage in the eyes of glaucoma patients. Am J Med. 114:638–646. 2003. View Article : Google Scholar : PubMed/NCBI

49 

Zhou B, Li C, Qi W, Zhang Y, Zhang F, Wu JX, Hu YN, Wu DM, Liu Y, Yan TT, et al: Downregulation of miR-181a upregulates sirtuin-1 (SIRT1) and improves hepatic insulin sensitivity. Diabetologia. 55:2032–2043. 2012. View Article : Google Scholar : PubMed/NCBI

50 

Muratsu-Ikeda S, Nangaku M, Ikeda Y, Tanaka T, Wada T and Inagi R: Downregulation of miR-205 modulates cell susceptibility to oxidative and endoplasmic reticulum stresses in renal tubular cells. PLoS One. 7:e414622012. View Article : Google Scholar : PubMed/NCBI

51 

Lee S, Yun I, Ham O, Lee SY, Lee CY, Park JH, Lee J, Seo HH, Choi E and Hwang KC: Suppression of miR-181a attenuates H2O2-induced death of mesenchymal stem cells by maintaining hexokinase II expression. Biol Res. 48:452015. View Article : Google Scholar : PubMed/NCBI

52 

Cao MQ, You AB, Zhu XD, Zhang W, Zhang YY, Zhang SZ, Zhang KW, Cai H, Shi WK, Li XL, et al: miR-182-5p promotes hepatocellular carcinoma progression by repressing FOXO3a. J Hematol Oncol. 11:122018. View Article : Google Scholar : PubMed/NCBI

53 

Wang D, Lu G, Shao Y and Xu D: MiR-182 promotes prostate cancer progression through activating Wnt/β-catenin signal pathway. Biomed Pharmacother. 99:334–339. 2018. View Article : Google Scholar : PubMed/NCBI

54 

Crow MT: Hypoxia, BNip3 proteins, and the mitochondrial death pathway in cardiomyocytes. Circ Res. 91:183–185. 2002. View Article : Google Scholar : PubMed/NCBI

55 

Regula KM, Ens K and Kirshenbaum LA: Inducible expression of BNIP3 provokes mitochondrial defects and hypoxia-mediated cell death of ventricular myocytes. Circ Res. 91:226–231. 2002. View Article : Google Scholar : PubMed/NCBI

56 

Zhang J, Ye J, Altafaj A, Cardona M, Bahi N, Llovera M, Cañas X, Cook SA, Comella JX and Sanchis D: EndoG links Bnip3-induced mitochondrial damage and caspase-independent DNA fragmentation in ischemic cardiomyocytes. PLoS One. 6:e179982011. View Article : Google Scholar : PubMed/NCBI

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August 2019
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Copy and paste a formatted citation
APA
Li, X., Wang, Q., Ren, Y., Wang, X., Cheng, H., Yang, H., & Wang, B. (2019). Tetramethylpyrazine protects retinal ganglion cells against H2O2‑induced damage via the microRNA‑182/mitochondrial pathway. International Journal of Molecular Medicine, 44, 503-512. https://doi.org/10.3892/ijmm.2019.4214
MLA
Li, X., Wang, Q., Ren, Y., Wang, X., Cheng, H., Yang, H., Wang, B."Tetramethylpyrazine protects retinal ganglion cells against H2O2‑induced damage via the microRNA‑182/mitochondrial pathway". International Journal of Molecular Medicine 44.2 (2019): 503-512.
Chicago
Li, X., Wang, Q., Ren, Y., Wang, X., Cheng, H., Yang, H., Wang, B."Tetramethylpyrazine protects retinal ganglion cells against H2O2‑induced damage via the microRNA‑182/mitochondrial pathway". International Journal of Molecular Medicine 44, no. 2 (2019): 503-512. https://doi.org/10.3892/ijmm.2019.4214