Polydatin protects against lipopolysaccharide-induced endothelial barrier disruption via SIRT3 activation

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Abstract

In a previous study, we demonstrated the role of polydatin (PD) in protecting against multiple organ dysfunction in sepsis. The aim of this study is to investigate whether PD protects against lipopolysaccharide (LPS)-induced endothelial barrier disruption through SIRT3 activation and to disclose the underlying mechanisms. Wild-type mice were injected with LPS and Evans Blue assay was performed to evaluate vascular permeability. Primary human umbilical vein endothelial cells (HUVECs) were stimulated with LPS. Endothelial permeability was evaluated by transendothelial electrical resistance (TER) and FITC-dextran leakage. SIRT3 activity was determined by a Deacetylase Fluorometric kit, and protein expression level of SIRT3 was detected by western blotting. Mitochondrial function was evaluated by determination of ROS level, mitochondrial membrane potential and mPTP opening. In endotoxemic mice, PD pretreatment attenuated vascular leakage in multiple organs while SIRT3 inhibition with 3-TYP reversed the effects of PD. PD treatment in late sepsis also exhibited barrier protective effects. In HUVECs, PD alleviated LPS-induced F-actin rearrangement, cadherin–catenin complex dissociation and endothelial hyperpermeability, whereas 3-TYP or SIRT3 siRNA attenuated the protective effects of PD. PD enhanced SIRT3 deacetylase activity, and attenuated LPS-induced decrease in SIRT3 expression as well. Furthermore, gain-of-function and loss-of-function strategies also confirmed the role of SIRT3 in enhancing endothelial barrier integrity. It was further ascertained that PD enhanced SIRT3-mediated deacetylation of SOD2 and cyclophilin D (CypD), thus suppressing mitochondrial dysfunction and subsequent endothelial barrier dysfunction. In addition, it was revealed that RAGE was involved in LPS-regulated SIRT3 signaling. Our results suggest that polydatin protects against LPS-induced endothelial barrier disruption dependent on SIRT3 and can be applied as a potential therapy for sepsis.

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References

  1. 1.

    Singer M, Deutschman CS, Seymour CW, Shankar-Hari M, Annane D, Bauer M, et al. The third international consensus definitions for sepsis and septic shock (Sepsis-3). JAMA. 2016;315:801–10.

  2. 2.

    De Backer D, Orbegozo Cortes D, Donadello K, Vincent JL. Pathophysiology of microcirculatory dysfunction and the pathogenesis of septic shock. Virulence. 2014;5:73–9.

  3. 3.

    Geven C, Kox M, Pickkers P. Adrenomedullin and adrenomedullin-targeted therapy as treatment strategies relevant for sepsis. Front Immunol. 2018;9:292.

  4. 4.

    Dejana E, Orsenigo F, Lampugnani MG. The role of adherens junctions and VE-cadherin in the control of vascular permeability. J Cell Sci. 2008;121(Pt 13):2115–22.

  5. 5.

    Mishra R, Singh SK. HIV-1 Tat C phosphorylates VE-cadherin complex and increases human brain microvascular endothelial cell permeability. BMC Neurosci. 2014;15:80.

  6. 6.

    Lee WL, Slutsky AS. Sepsis and endothelial permeability. N Engl J Med. 2010;363:689–91.

  7. 7.

    Zeng Z, Chen Z, Li T, Zhang J, Gao Y, Xu S, et al. Polydatin: a new therapeutic agent against multiorgan dysfunction. J Surg Res. 2015;198:192–9.

  8. 8.

    Xu S, Gao Y, Zhang Q, Wei S, Chen Z, Dai X, et al. SIRT1/3 activation by resveratrol attenuates acute kidney injury in a septic rat model. Oxid Med Cell Longev. 2016;2016:7296092.

  9. 9.

    Zeng Z, Yang Y, Dai X, Xu S, Li T, Zhang Q, et al. Polydatin ameliorates injury to the small intestine induced by hemorrhagic shock via SIRT3 activation-mediated mitochondrial protection. Exp Opin Ther Targets. 2016;20:645–52.

  10. 10.

    Ahn BH, Kim HS, Song S, Lee IH, Liu J, Vassilopoulos A, et al. A role for the mitochondrial deacetylase Sirt3 in regulating energy homeostasis. Proc Natl Acad Sci USA. 2008;105:14447–52.

  11. 11.

    Jing E, O'Neill BT, Rardin MJ, Kleinridders A, Ilkeyeva OR, Ussar S, et al. Sirt3 regulates metabolic flexibility of skeletal muscle through reversible enzymatic deacetylation. Diabetes. 2013;62:3404–17.

  12. 12.

    Samant SA, Zhang HJ, Hong Z, Pillai VB, Sundaresan NR, Wolfgeher D, et al. SIRT3 deacetylates and activates OPA1 to regulate mitochondrial dynamics during stress. Mol Cell Biol. 2014;34:807–19.

  13. 13.

    Sun F, Si Y, Bao H, Xu Y, Pan X, Zeng L, et al. Regulation of sirtuin 3-mediated deacetylation of cyclophilin D attenuated cognitive dysfunction induced by sepsis-associated encephalopathy in mice. Cell Mol Neurobiol. 2017;37:1457–64.

  14. 14.

    Zhao WY, Zhang L, Sui MX, Zhu YH, Zeng L. Protective effects of sirtuin 3 in a murine model of sepsis-induced acute kidney injury. Sci Rep. 2016;6:33201.

  15. 15.

    Zeng H, He X, Tuo QH, Liao DF, Zhang GQ, Chen JX. LPS causes pericyte loss and microvascular dysfunction via disruption of Sirt3/angiopoietins/Tie-2 and HIF-2alpha/Notch3 pathways. Sci Rep. 2016;6:20931.

  16. 16.

    Hafner AV, Dai J, Gomes AP, Xiao CY, Palmeira CM, Rosenzweig A, et al. Regulation of the mPTP by SIRT3-mediated deacetylation of CypD at lysine 166 suppresses age-related cardiac hypertrophy. Aging. 2010;2:914–23.

  17. 17.

    Li P, Meng X, Bian H, Burns N, Zhao KS, Song R. Activation of sirtuin 1/3 improves vascular hyporeactivity in severe hemorrhagic shock by alleviation of mitochondrial damage. Oncotarget. 2015;6:36998–7011.

  18. 18.

    Pi H, Xu S, Reiter RJ, Guo P, Zhang L, Li Y, et al. SIRT3-SOD2-mROS-dependent autophagy in cadmium-induced hepatotoxicity and salvage by melatonin. Autophagy. 2015;11:1037–51.

  19. 19.

    Qiu X, Brown K, Hirschey MD, Verdin E, Chen D. Calorie restriction reduces oxidative stress by SIRT3-mediated SOD2 activation. Cell Metab. 2010;12:662–7.

  20. 20.

    Wang L, Wu J, Guo X, Huang X, Huang Q. RAGE plays a role in LPS-induced NF-kappaB activation and endothelial hyperpermeability. Sensors. 2017;17. PMID: 28358333; https://doi.org/10.3390/s17040722.

  21. 21.

    Tinsley JH, Wu MH, Ma W, Taulman AC, Yuan SY. Activated neutrophils induce hyperpermeability and phosphorylation of adherens junction proteins in coronary venular endothelial cells. J Biol Chem. 1999;274:24930–4.

  22. 22.

    Gao Y, Zeng Z, Li T, Xu S, Wang X, Chen Z, et al. Polydatin inhibits mitochondrial dysfunction in the renal tubular epithelial cells of a rat model of sepsis-induced acute kidney injury. Anesth Analg. 2015;121:1251–60.

  23. 23.

    Chen T, Dai SH, Li X, Luo P, Zhu J, Wang YH, et al. Sirt1-Sirt3 axis regulates human blood–brain barrier permeability in response to ischemia. Redox Biol. 2018;14:229–36.

  24. 24.

    Chen ML, Zhu XH, Ran L, Lang HD, Yi L, Mi MT. Trimethylamine-N-oxide induces vascular inflammation by activating the NLRP3 inflammasome through the SIRT3-SOD2-mtROS signaling pathway. J Am Heart Assoc. 2017;6. PMID: 29118033; https://doi.org/10.1161/JAHA.117.002238.

  25. 25.

    Cheng A, Yang Y, Zhou Y, Maharana C, Lu D, Peng W, et al. Mitochondrial SIRT3 mediates adaptive responses of neurons to exercise and metabolic and excitatory challenges. Cell Metab. 2016;23:128–42.

  26. 26.

    Zeng X, Yang J, Hu O, Huang J, Ran L, Chen M, et al. Dihydromyricetin ameliorates nonalcoholic fatty liver disease by improving mitochondrial respiratory capacity and redox homeostasis through modulation of SIRT3 signaling. Antioxid Redox Signal. 2019;30:163–83.

  27. 27.

    Song C, Zhao J, Fu B, Li D, Mao T, Peng W, et al. Melatonin-mediated upregulation of Sirt3 attenuates sodium fluoride-induced hepatotoxicity by activating the MT1-PI3K/AKT-PGC-1alpha signaling pathway. Free Radic Biol Med. 2017;112:616–30.

  28. 28.

    Parodi-Rullan RM, Chapa-Dubocq X, Rullan PJ, Jang S, Javadov S. High sensitivity of SIRT3 deficient hearts to ischemia-reperfusion is associated with mitochondrial abnormalities. Front Pharmacol. 2017;8:275.

  29. 29.

    Chang M, Zhang B, Tian Y, Hu M, Zhang G, Di Z, et al. AGEs decreased SIRT3 expression and SIRT3 activation protected AGEs-induced EPCs' dysfunction and strengthened anti-oxidant capacity. Inflammation. 2017;40:473–85.

  30. 30.

    Liu TF, Vachharajani V, Millet P, Bharadwaj MS, Molina AJ, McCall CE. Sequential actions of SIRT1-RELB-SIRT3 coordinate nuclear-mitochondrial communication during immunometabolic adaptation to acute inflammation and sepsis. J Biol Chem. 2015;290:396–408.

  31. 31.

    Zheng Z, Ma H, Zhang X, Tu F, Wang X, Ha T, et al. Enhanced glycolytic metabolism contributes to cardiac dysfunction in polymicrobial sepsis. J Infect Dis. 2017;215:1396–406.

  32. 32.

    Pillai VB, Kanwal A, Fang YH, Sharp WW, Samant S, Arbiser J, et al. Honokiol, an activator of Sirtuin-3 (SIRT3) preserves mitochondria and protects the heart from doxorubicin-induced cardiomyopathy in mice. Oncotarget. 2017;8:34082–98.

  33. 33.

    Zhai M, Li B, Duan W, Jing L, Zhang B, Zhang M, et al. Melatonin ameliorates myocardial ischemia reperfusion injury through SIRT3-dependent regulation of oxidative stress and apoptosis. J Pineal Res. 2017;63. PMID: 28500761; https://doi.org/10.1111/jpi.12419

  34. 34.

    Zeng Z, Chen Z, Xu S, Song R, Yang H, Zhao KS. Polydatin alleviates small intestine injury during hemorrhagic shock as a SIRT1 activator. Oxid Med Cell Longev. 2015;2015:965961.

  35. 35.

    Vestweber D, Winderlich M, Cagna G, Nottebaum AF. Cell adhesion dynamics at endothelial junctions: VE-cadherin as a major player. Trends Cell Biol. 2009;19:8–15.

  36. 36.

    Fang D, Hawke D, Zheng Y, Xia Y, Meisenhelder J, Nika H, et al. Phosphorylation of beta-catenin by AKT promotes beta-catenin transcriptional activity. J Biol Chem. 2007;282:11221–9.

  37. 37.

    Dong WW, Liu YJ, Lv Z, Mao YF, Wang YW, Zhu XY, et al. Lung endothelial barrier protection by resveratrol involves inhibition of HMGB1 release and HMGB1-induced mitochondrial oxidative damage via an Nrf2-dependent mechanism. Free Radic Biol Med. 2015;88(Pt B):404–16.

  38. 38.

    Tharakan B, Holder-Haynes JG, Hunter FA, Smythe WR, Childs EW. Cyclosporine A prevents vascular hyperpermeability after hemorrhagic shock by inhibiting apoptotic signaling. J Trauma. 2009;66:1033–9.

  39. 39.

    Fonai F, Priber JK, Jakus PB, Kalman N, Antus C, Pollak E, et al. Lack of cyclophilin D protects against the development of acute lung injury in endotoxemia. Biochim Biophys Acta. 2015;1852:2563–73.

  40. 40.

    Kurundkar D, Kurundkar AR, Bone NB, Becker EJ, Jr., Liu W, Chacko B, et al. SIRT3 diminishes inflammation and mitigates endotoxin-induced acute lung injury. JCI Insight. 2019;4.

  41. 41.

    Schwer B, Bunkenborg J, Verdin RO, Andersen JS, Verdin E. Reversible lysine acetylation controls the activity of the mitochondrial enzyme acetyl-CoA synthetase 2. Proc Natl Acad Sci USA. 2006;103:10224–9.

  42. 42.

    Sundaresan NR, Gupta M, Kim G, Rajamohan SB, Isbatan A, Gupta MP. Sirt3 blocks the cardiac hypertrophic response by augmenting Foxo3a-dependent antioxidant defense mechanisms in mice. J Clin Investig. 2009;119:2758–71.

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Funding

This work was supported by the National Natural Science Foundation of China (81870210, 81170297, 81871604, 81701955, and 81703935), the Guangdong Natural Science Foundation Team Project (S2013030013217), the Natural Science Foundation of Guangdong Province (2016A030313561, 2016A030310389, and 2017A030313590), the Medical Science and Technology Research Fund of Guangdong Province (A2019178), and the Outstanding Youths Development Scheme of Nanfang Hospital, Southern Medical University (2016J011).

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Correspondence to Qiaobing Huang or Zhongqing Chen.

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Wu, J., Deng, Z., Sun, M. et al. Polydatin protects against lipopolysaccharide-induced endothelial barrier disruption via SIRT3 activation. Lab Invest (2019). https://doi.org/10.1038/s41374-019-0332-8

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