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Genipin inhibits the scleral expression of miR-29 and MMP2 and promotes COL1A1 expression in myopic eyes of guinea pigs

  • Basic Science
  • Published:
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Abstract

Purpose

High myopia can lead to blindness. Genipin is a collagen cross-linking agent that may be used to treat myopia. However, the mechanism of action of genipin for the treatment of myopia is unclear. This study investigated the effect of genipin on the scleral expression of the miR-29 cluster, matrix metalloproteinase 2 (MMP2), and collagen alpha1 chain of type I (COL1A1) in a guinea pig model of myopia.

Methods

The model of myopia was established by treating guinea pigs with a − 8D lens on both eyes for 21 days, and eyes with a refractive error of − 6D or greater were included. Quantitative reverse transcription–polymerase chain reaction (RT-PCR) and western blot were used to examine the mRNA and protein expression, respectively. A dual-luciferase assay was used to determine the direct targeting of the miR-29 cluster on the 3′-untranslated region (UTR) of the COL1A1 gene.

Results

The scleral expression of miR-29a, miR-29b, and miR-29c as well as MMP2 was significantly increased, and the scleral expression of COL1A1 was significantly decreased in the myopia group. Genipin treatment reversed these effects in myopic eyes. The dual-luciferase assay showed that the luciferase activities were significantly decreased in human embryonic kidney (HEK) cells transfected with miR-29a and miR-29b, but not miR-29c, compared with those transfected with control miRNAs.

Conclusions

Genipin inhibits the scleral expression of the miR-29 cluster and MMP2 and promotes COL1A1 expression in a guinea pig model of myopia. Thus, genipin may promote COL1A1 expression by reducing the expression of the miR-29 cluster.

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Availability of data and materials

The datasets generated and analyzed during the present study are available from the corresponding author on reasonable request.

Abbreviations

COL1A1:

Collagen alpha1 chain of type I

RT-PCR:

Reverse transcription–polymerase chain reaction

UTR:

Untranslated region

HEK:

Human embryonic kidney

MMP2:

Matrix metalloproteinase 2

miRNAs:

MicroRNAs

References

  1. McBrien NA, Gentle A (2003) Role of the sclera in the development and pathological complications of myopia. Prog Retin Eye Res 22:307–338

    Article  CAS  Google Scholar 

  2. Li J, Zhang Q (2017) Insight into the molecular genetics of myopia. Mol Vis 23:1048–1080

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Leo SW, Young TL (2011) An evidence-based update on myopia and interventions to retard its progression. J AAPOS 15:181–189. https://doi.org/10.1016/j.jaapos.2010.09.020

    Article  PubMed  PubMed Central  Google Scholar 

  4. Norton TT, Rada JA (1995) Reduced extracellular matrix in mammalian sclera with induced myopia. Vis Res 35:1271–1281

    Article  CAS  Google Scholar 

  5. Harper AR, Summers JA (2015) The dynamic sclera: extracellular matrix remodeling in normal ocular growth and myopia development. Exp Eye Res 133:100–111. https://doi.org/10.1016/j.exer.2014.07.015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Liu TX, Wang Z (2017) Biomechanics of sclera crosslinked using genipin in rabbit. Int J Ophthalmol 10:355–360. https://doi.org/10.18240/ijo.2017.03.05

    Article  PubMed  PubMed Central  Google Scholar 

  7. Campbell IC, Hannon BG, Read AT, Sherwood JM, Schwaner SA, Ethier CR (2017) Quantification of the efficacy of collagen cross-linking agents to induce stiffening of rat sclera. J R Soc Interface 14. https://doi.org/10.1098/rsif.2017.0014

  8. Liu TX, Wang Z (2013) Collagen crosslinking of porcine sclera using genipin. Acta Ophthalmol 91:e253–e257. https://doi.org/10.1111/aos.12172

    Article  CAS  PubMed  Google Scholar 

  9. Zhu SQ, Zheng LY, Pan AP, Yu AY, Wang QM, Xue AQ (2016) The efficacy and safety of posterior scleral reinforcement using genipin cross-linked sclera for macular detachment and retinoschisis in highly myopic eyes. Br J Ophthalmol 100:1470–1475. https://doi.org/10.1136/bjophthalmol-2015-308087

    Article  PubMed  Google Scholar 

  10. Wang M, Corpuz CC (2015) Effects of scleral cross-linking using genipin on the process of form-deprivation myopia in the Guinea pig: a randomized controlled experimental study. BMC Ophthalmol 15:89. https://doi.org/10.1186/s12886-015-0086-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Wang Q, Zhao G, Xing S, Zhang L, Yang X (2011) Role of bone morphogenetic proteins in form-deprivation myopia sclera. Mol Vis 17:647–657

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Frost MR, Norton TT (2007) Differential protein expression in tree shrew sclera during development of lens-induced myopia and recovery. Mol Vis 13:1580–1588

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Gong B, Qu C, Huang XF, Ye ZM, Zhang DD, Shi Y, Chen R, Liu YP, Shuai P (2016) Genetic association of COL1A1 polymorphisms with high myopia in Asian population: a meta-analysis. Int J Ophthalmol 9:1187–1193. https://doi.org/10.18240/ijo.2016.08.16

    Article  PubMed  PubMed Central  Google Scholar 

  14. Zhang X, Zhou X, Qu X (2015) Association between COL1A1 polymorphisms and high myopia: a meta-analysis. Int J Clin Exp Med 8:5862–5868

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Yu L, Li ZK, Gao JR, Liu JR, Xu CT (2011) Epidemiology, genetics and treatments for myopia. Int J Ophthalmol 4:658–669. https://doi.org/10.3980/j.issn.2222-3959.2011.06.17

    Article  PubMed  PubMed Central  Google Scholar 

  16. Rada JA, Brenza HL (1995) Increased latent gelatinase activity in the sclera of visually deprived chicks. Invest Ophthalmol Vis Sci 36:1555–1565

    CAS  PubMed  Google Scholar 

  17. Rada JA, Perry CA, Slover ML, Achen VR (1999) Gelatinase a and TIMP-2 expression in the fibrous sclera of myopic and recovering chick eyes. Invest Ophthalmol Vis Sci 40:3091–3099

    CAS  PubMed  Google Scholar 

  18. Guggenheim JA, McBrien NA (1996) Form-deprivation myopia induces activation of scleral matrix metalloproteinase-2 in tree shrew. Invest Ophthalmol Vis Sci 37:1380–1395

    CAS  Google Scholar 

  19. Liu Z, Qiu F, Li J, Zhu Z, Yang W, Zhou X, An J, Huang F, Wang Q, Reinach PS, Li W, Chen W, Liu Z (2015) Adenomatous polyposis coli mutation leads to myopia development in mice. PLoS One 10:e0141144. https://doi.org/10.1371/journal.pone.0141144

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Zhou X, Ji F, An J, Zhao F, Shi F, Huang F, Li Y, Jiao S, Yan D, Chen X, Chen J, Qu J (2012) Experimental murine myopia induces collagen type Ialpha1 (COL1A1) DNA methylation and altered COL1A1 messenger RNA expression in sclera. Mol Vis 18:1312–1324

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Jiang B, Huo Y, Gu Y, Wang J (2017) The role of microRNAs in myopia. Graefes Arch Clin Exp Ophthalmol 255:7–13. https://doi.org/10.1007/s00417-016-3532-6

    Article  CAS  PubMed  Google Scholar 

  22. Luna C, Li G, Qiu J, Epstein DL, Gonzalez P (2009) Role of miR-29b on the regulation of the extracellular matrix in human trabecular meshwork cells under chronic oxidative stress. Mol Vis 15:2488–2497

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Xie M, Li Y, Wu J, Wu J (2016) Genetic variants in MiR-29a associated with high myopia. Ophthalmic Genet 37:456–458. https://doi.org/10.3109/13816810.2015.1101776

    Article  PubMed  Google Scholar 

  24. Wang M, Yang ZK, Li RQ, Fan H, Liu Y (2019) Comparison of the biomechanics and fixation index of crosslinking between lysyl oxidase and genipin on guinea pig sclera, an animal model of defocus-induced high myopia. Proc Est Acad Sci 2:214–221

    Article  CAS  Google Scholar 

  25. McBrien NA, Cornell LM, Gentle A (2001) Structural and ultrastructural changes to the sclera in a mammalian model of high myopia. Invest Ophthalmol Vis Sci 42:2179–2187

    CAS  PubMed  Google Scholar 

  26. Vannini I, Fanini F, Fabbri M (2018) Emerging roles of microRNAs in cancer. Curr Opin Genet Dev. https://doi.org/10.1016/j.gde.2018.01.001

  27. Maurer B, Stanczyk J, Jungel A, Akhmetshina A, Trenkmann M, Brock M, Kowal-Bielecka O, Gay RE, Michel BA, Distler JH, Gay S, Distler O (2010) MicroRNA-29, a key regulator of collagen expression in systemic sclerosis. Arthritis Rheum 62:1733–1743. https://doi.org/10.1002/art.27443

    Article  CAS  PubMed  Google Scholar 

  28. Kim JW, Lindsey JD, Wang N, Weinreb RN (2001) Increased human scleral permeability with prostaglandin exposure. Invest Ophthalmol Vis Sci 42:1514–1521

    CAS  PubMed  Google Scholar 

  29. Wojciechowski R, Bailey-Wilson JE, Stambolian D (2010) Association of matrix metalloproteinase gene polymorphisms with refractive error in Amish and Ashkenazi families. Invest Ophthalmol Vis Sci 51:4989–4995. https://doi.org/10.1167/iovs.10-5474

    Article  PubMed  PubMed Central  Google Scholar 

  30. Nakanishi H, Hayashi H, Yamada R, Yamashiro K, Nakata I, Shimada N, Ohno-Matsui K, Mochizuki M, Ozaki M, Yoshitake S, Kuriyama S, Saito M, Iida T, Matsuo K, Matsuda F, Yoshimura N (2010) Single-nucleotide polymorphisms in the promoter region of matrix metalloproteinase-1, −2, and −3 in Japanese with high myopia. Invest Ophthalmol Vis Sci 51:4432–4436. https://doi.org/10.1167/iovs.09-4871

    Article  PubMed  Google Scholar 

  31. Leung KH, Yiu WC, Yap MK, Ng PW, Fung WY, Sham PC, Yip SP (2011) Systematic investigation of the relationship between high myopia and polymorphisms of the MMP2, TIMP2, and TIMP3 genes by a DNA pooling approach. Invest Ophthalmol Vis Sci 52:3893–3900. https://doi.org/10.1167/iovs.11-7286

    Article  CAS  PubMed  Google Scholar 

  32. Gong B, Liu X, Zhang D, Wang P, Huang L, Lin Y, Lu F, Ma S, Cheng J, Chen R, Li X, Lin H, Zeng G, Zhu X, Hu J, Yang Z, Shi Y (2013) Evaluation of MMP2 as a candidate gene for high myopia. Mol Vis 19:121–127

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Siegwart JT Jr, Norton TT (2005) Selective regulation of MMP and TIMP mRNA levels in tree shrew sclera during minus lens compensation and recovery. Invest Ophthalmol Vis Sci 46:3484–3492. https://doi.org/10.1167/iovs.05-0194

    Article  PubMed  PubMed Central  Google Scholar 

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Funding

Supported by National Natural Science Foundation of China (No.81770958).

Author information

Authors and Affiliations

  1. Aier School of Ophthalmology, Central South University, Changsha, 410015, China

    Min Wang, Zhi-Kuan Yang, Hong Liu & Rui-qin Li

  2. Shanghai Aier Eye Hospital, Shanghai, 200336, China

    Min Wang, Yu Liu & Wen-Jun Zhong

  3. Aier Institute of Optometry & Vision Science, Changsha, 410015, China

    Zhi-Kuan Yang

Authors
  1. Min Wang
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  2. Zhi-Kuan Yang
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  3. Hong Liu
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  4. Rui-qin Li
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  5. Yu Liu
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Contributions

Min Wang and Zhi-Kuan Yang designed most of the investigation, performed data analysis, and wrote the manuscript; Hong Liu provided pathological and experimental assistance; Rui-qin Li contributed to interpretation of the data and analyses. Yu Liu and Wen-Jun Zhong provided help with retinoscopy.

Corresponding author

Correspondence to Zhi-Kuan Yang.

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The authors declare that they have no conflicts of interest.

Research involving animals

All experimental protocols were approved by the Ethics Committee of the Shanghai Experimental Animal Society (Approval number: SZY201704007).

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Wang, M., Yang, ZK., Liu, H. et al. Genipin inhibits the scleral expression of miR-29 and MMP2 and promotes COL1A1 expression in myopic eyes of guinea pigs. Graefes Arch Clin Exp Ophthalmol 258, 1031–1038 (2020). https://doi.org/10.1007/s00417-020-04634-7

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  • DOI: https://doi.org/10.1007/s00417-020-04634-7

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