Inflammation Research

, Volume 66, Issue 2, pp 157–166 | Cite as

Activation of the TXNIP/NLRP3 inflammasome pathway contributes to inflammation in diabetic retinopathy: a novel inhibitory effect of minocycline

  • Wei Chen
  • Minjie Zhao
  • Shuzhi Zhao
  • Qianyi Lu
  • Lisha Ni
  • Chen Zou
  • Li Lu
  • Xun Xu
  • Huaijin Guan
  • Zhi ZhengEmail author
  • Qinghua QiuEmail author
Original Research Paper


Objective and design

Chronic low-grade inflammation occurs in diabetic retinopathy (DR), but the underlying mechanism(s) remains (remain) unclear. NLRP3 inflammasome activation is involved in several other inflammatory diseases. Thus, we investigated the role of the NLRP3 inflammasome signaling pathway in the pathogenesis of DR.


Diabetes was induced in rats by streptozotocin treatment for 8 weeks. They were treated with NLRP3 shRNA or minocycline during the last 4 weeks. High glucose-exposed human retinal microvascular endothelial cells (HRMECs) were co-incubated with antioxidants or subjected to TXNIP or NLRP3 shRNA interference.


In high glucose-exposed HRMECs and retinas of diabetic rats, mRNA and protein expression of NLRP3, ASC, and proinflammatory cytokines were induced significantly by hyperglycemia. Upregulated interleukin (IL)-1β maturation, IL-18 secretion, and caspase-1 cleavage were also observed with increased cell apoptosis and retinal vascular permeability, compared with the control group. NLRP3 silencing blocked these effects in the rat model and HRMECs, confirming that inflammasome activation contributed to inflammation in DR. TXNIP expression was increased by reactive oxygen species (ROS) overproduction in animal and cell models, whereas antioxidant addition or TXNIP silencing blocked IL-1β and IL-18 secretion in high glucose-exposed HRMECs, indicating that the ROS–TXNIP pathway mediates NLRP3 inflammasome activation. Minocycline significantly downregulated ROS generation and reduced TXNIP expression, subsequently inhibited NLRP3 activation, and further decreased inflammatory factors, which were associated with a decrease in retinal vascular permeability and cell apoptosis.


Together, our data suggest that the TXNIP/NLRP3 pathway is a potential therapeutic target for the treatment of DR, and the use of minocycline specifically for such therapy may be a new avenue of investigation in inflammatory disease.


NLRP3 inflammasome Diabetic retinopathy Minocycline 



This work was supported by grants from the National Nature Science Foundation of China (81271032) and the Shanghai Nature Science Foundation (No. 14ZR1433600).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.


  1. 1.
    Ding J, Wong TY. Current epidemiology of diabetic retinopathy and diabetic macular edema. Curr Diab Rep. 2012;12:346–54.CrossRefPubMedGoogle Scholar
  2. 2.
    El-Asrar AM. Role of inflammation in the pathogenesis of diabetic retinopathy. Middle East Afr J Ophthalmol. 2012;19:70–4.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kowluru RA, Odenbach S. Role of interleukin-1beta in the pathogenesis of diabetic retinopathy. Br J Ophthalmol. 2004;88:1343–7.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Martinon F, Burns K, Tschopp J. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol Cell. 2002;10:417–26.CrossRefPubMedGoogle Scholar
  5. 5.
    Cassel SL, Joly S, Sutterwala FS. The NLRP3 inflammasome: a sensor of immune danger signals. Semin Immunol. 2009;21:194–8.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Leavy O. Inflammasome: turning on and off NLRP3. Nat Rev Immunol. 2013;13:1.CrossRefPubMedGoogle Scholar
  7. 7.
    Michaudel C, Couturier-Maillard A, Chenuet P, et al. Inflammasome, IL-1 and inflammation in ozone-induced lung injury. Am J Clin Exp Immunol. 2016;5:33–40.PubMedPubMedCentralGoogle Scholar
  8. 8.
    Perrone L, Devi TS, Hosoya K, Terasaki T, Singh LP. Thioredoxin interacting protein (TXNIP) induces inflammation through chromatin modification in retinal capillary endothelial cells under diabetic conditions. J Cell Physiol. 2009;221:262–72.CrossRefPubMedGoogle Scholar
  9. 9.
    Zhou R, Tardivel A, Thorens B, Choi I, Tschopp J. Thioredoxin-interacting protein links oxidative stress to inflammasome activation. Nat Immunol. 2010;11:136–40.CrossRefPubMedGoogle Scholar
  10. 10.
    Perrone L, Devi TS, Hosoya KI, Terasaki T, Singh LP. Inhibition of TXNIP expression in vivo blocks early pathologies of diabetic retinopathy. Cell Death Dis. 2010;1:e65.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Li C, Yuan K, Schluesener H. Impact of minocycline on neurodegenerative diseases in rodents: a meta-analysis. Rev Neurosci. 2013;24:553–62.CrossRefPubMedGoogle Scholar
  12. 12.
    Krady JK, Basu A, Allen CM, Xu Y, LaNoue KF, Gardner TW, et al. Minocycline reduces proinflammatory cytokine expression, microglial activation, and caspase-3 activation in a rodent model of diabetic retinopathy. Diabetes. 2005;54:1559–65.CrossRefPubMedGoogle Scholar
  13. 13.
    Zheng Z, Chen H, Li J, Li T, Zheng B, Zheng Y, et al. Sirtuin 1-mediated cellular metabolic memory of high glucose via the LKB1/AMPK/ROS pathway and therapeutic effects of metformin. Diabetes. 2012;61:217–28.CrossRefPubMedGoogle Scholar
  14. 14.
    Leal EC, Martins J, Voabil P, Liberal J, Chiavaroli C, Bauer J, et al. Calcium dobesilate inhibits the alterations in tight junction proteins and leukocyte adhesion to retinal endothelial cells induced by diabetes. Diabetes. 2010;59:2637–45.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Kingsbury SR, Conaghan PG, McDermott MF. The role of the NLRP3 inflammasome in gout. J Inflamm Res. 2011;4:39–49.PubMedPubMedCentralGoogle Scholar
  16. 16.
    Lee HM, Kim JJ, Kim HJ, Shong M, Ku BJ, Jo EK. Upregulated NLRP3 inflammasome activation in patients with type 2 diabetes. Diabetes. 2013;62:194–204.CrossRefPubMedGoogle Scholar
  17. 17.
    Al-Shabrawey M, Zhang W, McDonald D. Diabetic retinopathy: mechanism, diagnosis, prevention, and treatment. BioMed Res Int. 2015;2015:854593.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Sagulenko V, Thygesen SJ, Sester DP, et al. AIM2 and NLRP3 inflammasomes activate both apoptotic and pyroptotic death pathways via ASC. Cell Death Differ. 2013;20:1149–60.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Shimada K, Crother TR, Karlin J, et al. Oxidized mitochondrial DNA activates the NLRP3 inflammasome during apoptosis. Immunity. 2012;36:401–14.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Heid ME, Keyel PA, Kamga C, Shiva S, Watkins SC, Salter RD. Mitochondrial reactive oxygen species induces NLRP3-dependent lysosomal damage and inflammasome activation. J Immunol. 2013;191:5230–8.CrossRefPubMedGoogle Scholar
  21. 21.
    Tschopp J, Schroder K. NLRP3 inflammasome activation: the convergence of multiple signalling pathways on ROS production? Nat Rev Immunol. 2010;10:210–5.CrossRefPubMedGoogle Scholar
  22. 22.
    Zhou R, Tardivel A, Thorens B, Choi I, Tschopp J. Thioredoxin-interacting protein links oxidative stress to inflammasome activation. Nat Immunol. 2009;11:136–40.CrossRefPubMedGoogle Scholar
  23. 23.
    Devi TS, Hosoya K-I, Terasaki T, Singh LP. Critical role of TXNIP in oxidative stress. DNA damage and retinal pericyte apoptosis under high glucose: Implications for diabetic retinopathy. Experimental cell research; 2013.Google Scholar
  24. 24.
    Szeto GL, Pomerantz JL, Graham DR, Clements JE. Minocycline suppresses activation of nuclear factor of activated T cells 1 (NFAT1) in human CD4+ T cells. J Biol Chem. 2011;286:11275–82.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Giuliani F, Hader W, Yong VW. Minocycline attenuates T cell and microglia activity to impair cytokine production in T cell–microglia interaction. J Leukoc Biol. 2005;78:135–43.CrossRefPubMedGoogle Scholar
  26. 26.
    Kraus RL, Pasieczny R, Lariosa-Willingham K, Turner MS, Jiang A, Trauger JW. Antioxidant properties of minocycline: neuroprotection in an oxidative stress assay and direct radical-scavenging activity. J Neurochem. 2005;94:819–27.CrossRefPubMedGoogle Scholar
  27. 27.
    Plane JM, Shen Y, Pleasure DE, Deng W. Prospects for minocycline neuroprotection. Arch Neurol. 2010;67:1442–8.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Bhatt LK, Addepalli V. Attenuation of diabetic retinopathy by enhanced inhibition of MMP-2 and MMP-9 using aspirin and minocycline in streptozotocin-diabetic rats. Am J Transl Res. 2010;2:181–9.PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing 2016

Authors and Affiliations

  • Wei Chen
    • 1
    • 2
  • Minjie Zhao
    • 1
  • Shuzhi Zhao
    • 1
  • Qianyi Lu
    • 1
  • Lisha Ni
    • 3
  • Chen Zou
    • 1
  • Li Lu
    • 4
  • Xun Xu
    • 1
  • Huaijin Guan
    • 2
  • Zhi Zheng
    • 1
    Email author
  • Qinghua Qiu
    • 1
    Email author
  1. 1.Department of Ophthalmology, Shanghai First People’s Hospital, School of MedicineShanghai Jiaotong UniversityShanghaiChina
  2. 2.Department of OphthalmologyAffiliated Hospital of Nantong UniversityNantongChina
  3. 3.Department of Ophthalmology, Lishui People’s HospitalLishuiChina
  4. 4.Department of OphthalmologyAnhui Provincial HospitalHefeiChina

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