Skip to main content
SpringerLink
Log in

Effects of electroacupuncture on conjunctival cell apoptosis and the expressions of apoptosis-related proteins Caspase-3, Fas and Bcl-2 in rabbits with dry eye syndrome

电针对干眼症兔结膜细胞凋亡及相关蛋白 Caspase-3, Fas和Bcl-2表达的影响

  • Basic Study
  • Published:
Journal of Acupuncture and Tuina Science Aims and scope Submit manuscript

Abstract

Objective

By observing the effects of electroacupuncture (EA) on the apoptosis of conjunctival cells of rabbits with dry eye syndrome (DES) and the expressions of apoptosis-related proteins Caspase-3, Fas and Bcl-2, to discuss the mechanism of EA in the treatment of DES from the perspective of cell apoptosis.

Methods

Male New Zealand rabbits were randomly divided into a normal group (NG), a model group (MG), an EA group (EAG) and a sham EA group (SEAG). DES rabbit model was developed by eye drop of 0.1% benzalkonium chloride. The rabbit tear secretion and tear film break-up time (BUT) were measured; terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect the apoptosis of conjunctival cells; the expressions of Caspase-3, Fas and Bcl-2 proteins in conjunctival cells were detected by immunohistochemistry.

Results

Compared with the NG, the rabbit tear secretion decreased and the BUT was shortened in the MG (both P<0.01); compared with the MG and the SEAG, the rabbit tear secretion increased and the BUT was prolonged in the EAG (all P<0.05). Compared with the NG, the apoptosis of rabbit conjunctival cells increased (P<0.01), the expressions of Caspase-3 and Fas proteins increased (both P<0.05), and the expression of Bcl-2 protein decreased (P<0.01) in the MG; compared with the MG and the SEAG, the apoptosis of rabbit conjunctival cells decreased (both P<0.01), the expressions of Caspase-3 and Fas proteins decreased (all P<0.05), and the expression of Bcl-2 protein increased (both P<0.01) in the EAG.

Conclusion

EA can inhibit the apoptosis of rabbit conjunctival cells, down-regulate the expressions of apoptosis-related proteins Caspase-3 and Fas, and up-regulate the expression of Bcl-2 protein, which may be one of the mechanisms of EA in treatment of DES.

摘要

目的

通过观察电针对干眼症兔结膜细胞凋亡及相关蛋白Caspase-3, Fas和Bcl-2表达的影响, 从细胞凋亡角度探讨电针治疗干眼症的作用机制.

方法

雄性新西兰兔随机分为正常组,模型组,电针组和假电针组. 采用0.1%苯扎氯铵滴眼制作兔干眼症模型. 检测各组兔泪液分泌量,泪膜破裂时间; 采用末端脱氧核苷酸转移酶介导的dUTP 缺口末端标记测定(TUNEL)法检测结膜细胞凋亡情况; 应用免疫组化法检测结膜细胞Caspase-3, Fas和 Bcl-2蛋白表达.

结果

与正常组比较, 模型组兔泪液分泌量降低, 泪膜破裂时间缩短(均P<0.01); 与模型组, 假电针组比较, 电针组兔泪液分泌量增多, 泪膜破裂时间延长(均P<0.05).与正常组比较, 模型组兔结膜细胞凋亡增加(P<0.01), Caspase-3和Fas蛋白表达增加(均P<0.05), Bcl-2蛋白表达降低(P<0.01); 与模型组,假电针组比较, 电针组兔结膜细胞凋亡减少(均P<0.01), Caspase-3和Fas蛋白表达减少(均P<0.05), Bcl-2蛋白表达增加(均P<0.01).

结论

电针能抑制干眼症兔结膜细胞凋亡, 下调凋亡相关蛋白Caspase-3和Fas表达, 上调Bcl-2蛋白表达, 该作用可能是电针治疗干眼症的作用机制之一.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Craig JP, Nelson JD, Azar DT, Belmonte C, Bron AJ, Chauhan SK, de Paiva CS, Gomes JAP, Hammitt KM, Jones L, Nichols JJ, Nichols KK, Novack GD, Stapleton FJ, Willcox MDP, Wolffsohn JS, Sullivan DA. TFOS DEWS II report executive summary. Ocul Surf, 2017, 15(4): 802–812.

    Article  Google Scholar 

  2. Malik K, Matejtschuk P, Thelwell C, Burns CJ. Differential scanning fluorimetry: rapid screening of formulations that promote the stability of reference preparations. J Pharm Biomed Anal, 2013, 77: 163–166.

    Article  CAS  Google Scholar 

  3. Than J, Balal S, Wawrzynski J, Nesaratnam N, Saleh GM, Moore J, Patel A, Shah S, Sharma B, Kumar B, Smith J, Sharma A. Fingerprick autologous blood: a novel treatment for dry eye syndrome. Eye, 2017, 31(12): 1655–1663.

    Article  CAS  Google Scholar 

  4. Galor A, Batawi H, Felix ER, Margolis TP, Sarantopoulos KD, Martin ER, Levitt RC. Incomplete response to artificial tears is associated with features of neuropathic ocular pain. Br J Ophthalmol, 2016, 100(6): 745–749.

    Article  Google Scholar 

  5. Kim BH, Kim MH, Kang SH, Nam HJ. Optimizing acupuncture treatment for dry eye syndrome: a systematic review. BMC Complement Altern Med, 2018, 18(1): 145.

    Article  Google Scholar 

  6. Yang L, Yang Z, Yu H, Song H. Acupuncture therapy is more effective than artificial tears for dry eye syndrome: evidence based on a meta-analysis. Evid Based Complement Alternat Med, 2015, 2015: 143858.

    PubMed  PubMed Central  Google Scholar 

  7. Brinton M, Chung JL, Kossler A, Kook KH, Loudin J, Franke M, Palanker D. Electronic enhancement of tear secretion. J Neural Eng, 2016, 13(1): 016006.

    Article  Google Scholar 

  8. Hang W, Li W, Zhang C, Zhu C, Yi X, Zhou Y, Lü Y. Effects of vitamin Aon expressions of apoptosis genes Bax and Bcl-2 in epithelial cells of corneal tissues induced by benzalkonium chloride in mice with dry eye. Med Sci Monit, 2019, 25: 4583–4589.

    Article  Google Scholar 

  9. Park JH, Moon SH, Kang DH, Um HJ, Kang SS, Kim JY, Tchah H. Diquafosol sodium inhibits apoptosis and inflammation of corneal epithelial cells via activation of Erk1/2 and RSK: in vitro and in vivo dry eye model. Invest Ophthalmol Vis Sci, 2018, 59(12): 5108–5115.

    Article  CAS  Google Scholar 

  10. Chen W, Zhang X, Liu M, Zhang J, Ye Y, Lin Y, Luyckx J, Qu J. Trehalose protects against ocular surface disorders in experimental murine dry eye through suppression of apoptosis. Exp Eye Res, 2009, 89(3): 311–318.

    Article  CAS  Google Scholar 

  11. Gao J, Schwalb TA, Addeo JV, Ghosn CR, Stern ME. The role of apoptosis in the pathogenesis of canine keratoconjunctivitis sicca: the effect of topical cyclosporin atherapy. Cornea, 1998, 17(6): 654–663.

    Article  CAS  Google Scholar 

  12. Li C, Song Y, Luan S, Wan P, Li N, Tang J, Han Y, Xiong C, Wang Z. Research on the stability of a rabbit dry eye model induced by topical application of the preservative benzalkonium chloride. PloS one, 2012, 7(3): e33688.

    Article  CAS  Google Scholar 

  13. Xiong C, Chen D, Liu J, Liu B, Li N, Zhou Y, Liang X, Ma P, Ye C, Ge J, Wang Z. A rabbit dry eye model induced by topical medication of a preservative benzalkonium chloride. Invest Ophthalmol Vis Sci, 2008, 49(5): 1850–1856.

    Article  Google Scholar 

  14. Wang CL, Cheng ZL, Li GL. Study on standardization of methods commonly used experimental animal acupoint location. Tianjin Zhongyiyao, 2016, 33(2): 100–103.

    Google Scholar 

  15. Li ZR. Experimental Acupuncture Science. Beijing: China Press of Traditional Chinese Medicine, 2007: 327–329.

    Google Scholar 

  16. Honkanen RA, Huang L, Xie G, Rigas B. Phosphosulindac is efficacious in an improved concanavalin A-based rabbit model of chronic dry eye disease. Transl Res, 2018, 198: 58–72.

    Article  CAS  Google Scholar 

  17. Xue Y, Zhao YD, Yuan YJ, Wang XF. Research progress in nearly five years of acupuncture in the treatment of dry eyes. Yatai Chuantong Yiyao, 2017, 13(3): 59–61.

    Google Scholar 

  18. Guo MH, Cui EC, Li XY, Zong L. Observations on the clinical therapeutic effect of electroacupuncture on xerophthalmia. Shanghai Zhenjiu Zazhi, 2012, 31(4): 245–247.

    Google Scholar 

  19. Baudouin C. The pathology of dry eye. Surv Ophthalmol, 2001, 45(2): S211–S220.

    Article  Google Scholar 

  20. Fernandez KB, Epstein SP, Raynor GS, Sheyman AT, Massingale ML, Dentone PG, Landegger LD, Asbell PA. Modulation of HLA-DR in dry eye patients following 30 days of treatment with a lubricant eyedrop solution. Clin Ophthalmology, 2015, 9: 1137–1145.

    CAS  Google Scholar 

  21. Yeh S, Song XJ, Farley W, Li DQ, Stern ME, Pflugfelder SC. Apoptosis of ocular surface cells in experimentally induced dry eye. Inves Ophthalmol Vis Sci, 2003, 44(1): 124–129.

    Article  Google Scholar 

  22. Hagar H, Husain S, Fadda LM, Attia NM, Attia MMA, Ali HM. Inhibition of NF-κB and the oxidative stress-dependent caspase-3 apoptotic pathway by betaine supplementation attenuates hepatic injury mediated by cisplatin in rats. Pharmacol Rep, 2019, 71(6): 1025–1033.

    Article  Google Scholar 

  23. Liu J, Yao Y, Ding H, Chen R. Oxymatrine triggers apoptosis by regulating Bcl-2 family proteins and activating caspase-3/caspase-9 pathway in human leukemia HL-60 cells. Tumor Biol, 2014, 35(6): 5409–5415.

    Article  CAS  Google Scholar 

  24. Wang CQ, Li D, Liu HP, Li L. Effects of Yangyin Runmu pills on the expressions of caspase-3 and caspase-6 in conjunctival epithelial cells in a rat model of dry eye. Hunan Zhongyiyao Daxue Xuebao, 2019, 39(7): 848–850.

    Google Scholar 

  25. Sun JH, Wang JX. Inhibition of cyclosporine on apoptosis of conjunctival epithelium in experimental murine xerophthalamia. Yanke Yanjiu, 2006, 24(5): 506–509.

    CAS  Google Scholar 

  26. Xie J, Li B, Yao B, Zhang P, Wang L, Lu H, Song X. Transforming growth factor-β1-regulated Fas/FasL pathway activation suppresses nucleus pulposus cell apoptosis in an inflammatory environment. Biosci Rep, 2019: BSR20191726.

  27. Yao XL, Peng QH, Chen QL, Tang YH, Zhong Q. Effects of total flavonoid of chrysanthemum on Fas and FasL expression in lacrimal gland cells of male castrated rabbits with dry eye. Guoji Yanke Zazhi, 2014, 14(10): 1749–1754.

    CAS  Google Scholar 

  28. Birkinshaw RW, Czabotar PE. The BCL-2 family of proteins and mitochondrial outer membrane permeabilisation. Semin Cell Dev Biol, 2017, 72: 152–162.

    Article  CAS  Google Scholar 

  29. Cong Y, Wang WM. Relationship between Fas, FasL, Bax and Bcl-2 expressions in dry eye corneal epithelial cells induced by vitamin A deficiency and apoptosis. Zhongguo Shiyan Zhenduanxue, 2017, 21(10): 1820–1823.

    Google Scholar 

  30. Ouyang Y, Peng J, Peng QH, Tan HY, Li WJ, Wu QL. Effect of buddleia granule on Bax, Bcl-2, Fas, and FasL in lacrimal gland cells of castrated male rabbits. Zhongguo Zhongxiyi Jiehe Zazhi, 2017, 37(7): 858–862.

    Google Scholar 

  31. Strong B, Farley W, Stern ME, Pflugfelder SC. Topical cyclosporine inhibits conjunctival epithelial apoptosis in experimental murine keratoconjunctivitis sicca. Cornea, 2005, 24(1): 80–85.

    Article  Google Scholar 

Download references

Acknowledgments

This work was sponsored by the Youth Fund of National Natural Science Foundation of China (国家自然科学基金青年基金项目, No. 81904302); Scientific Research Project of Shanghai Municipal Health Commission (上海市卫生健康委员会项目, No. 2017BR047, No. 201840174, No. 201940130); Shanghai Sailing Program (上海市青年科技英才扬帆计划项目, No. 19YF1445000).

Author information

Authors and Affiliations

  1. Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 200030, China

    Yan-ting Yang  (杨延婷), Yue Zhao  (赵越), Dan Zhang  (张丹), Jie Liu  (刘婕), Cui-hong Zhang  (张翠红), Ling-xiang Wu  (吴凌翔), Jue Hong  (洪珏) & Xiao-peng Ma  (马晓芃)

  2. Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

    Bang-ji Wei  (魏邦基), Xiao-qing Dong  (董小庆), Lin-lin Zhang  (张琳琳) & Xiao-peng Ma  (马晓芃)

  3. Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, 100053, China

    Xiao-xu Liu  (刘晓旭)

Authors
  1. Yan-ting Yang  (杨延婷)
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bang-ji Wei  (魏邦基)
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yue Zhao  (赵越)
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dan Zhang  (张丹)
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jie Liu  (刘婕)
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Cui-hong Zhang  (张翠红)
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ling-xiang Wu  (吴凌翔)
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiao-qing Dong  (董小庆)
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Xiao-xu Liu  (刘晓旭)
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Lin-lin Zhang  (张琳琳)
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Jue Hong  (洪珏)
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Xiao-peng Ma  (马晓芃)
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jue Hong  (洪珏) or Xiao-peng Ma  (马晓芃).

Additional information

Conflict of Interest

The authors declare that there is no potential conflict of interest in this article.

Statement of Human and Animal Rights

The treatment of animals conformed to the ethical criteria.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, Yt., Wei, Bj., Zhao, Y. et al. Effects of electroacupuncture on conjunctival cell apoptosis and the expressions of apoptosis-related proteins Caspase-3, Fas and Bcl-2 in rabbits with dry eye syndrome. J. Acupunct. Tuina. Sci. 18, 16–23 (2020). https://doi.org/10.1007/s11726-020-1152-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11726-020-1152-5

Keywords

关键词

中图分类号

文献标志码

Search

Navigation