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MALAT1 Up-Regulator Polydatin Protects Brain Microvascular Integrity and Ameliorates Stroke Through C/EBPβ/MALAT1/CREB/PGC-1α/PPARγ Pathway

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Abstract

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is a long non-coding RNA contributing to protect the blood–brain barrier (BBB) after stroke. We searched for small molecules that may up-regulate MALAT1 and focused on polydatin (PD), a natural product, as a possible candidate. PD enhanced MALAT1 gene expression in rat brain microvascular endothelial cells, reducing cell toxicity and apoptosis after oxygen and glucose deprivation (OGD). These effects correlated with reduction of inflammatory factors and enhancement of expression of BBB markers. We found opposite changes after MALAT1 silencing. We determined that C/EBPβ is a key transcription factor for PD-mediated MALAT1 expression. PPARγ activity is involved in MALAT1 protective effects through its coactivator PGC-1α and the transcription factor CREB. This suggests that PD activates the MALAT1/CREB/PGC-1α/PPARγ signaling pathway to protect endothelial cells against ischemia. PD administration to rats subjected to brain ischemia by transient middle cerebral artery occlusion (tMCAO) reduced cerebral infarct volume and brain inflammation, protected cerebrovascular endothelial cells and BBB integrity. These effects correlated with increased expression of MALAT1, C/EBPβ, and PGC-1α. Our results strongly suggest that the beneficial effects of PD involve the C/EBPβ/MALAT1/CREB/PGC-1α/PPARγ pathway, which may provide a novel therapeutic strategy for brain ischemic stroke.

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Abbreviations

ABCG1:

ATP-binding cassette sub-family G member 1

BBB:

Blood–brain barrier

CD36:

Cluster of differentiation 36

C/EBPβ:

CCAAT/enhancer-binding proteinβ

COX-2:

Cyclooxygenase-2

CREB:

cAMP response element binding

DMEM:

Dulbecco’s modified eagle’s medium

DMSO:

Dimethyl sulfoxide

FBS:

Fetal bovine serum

HUVEC:

Human umbilical vein endothelial cells

IL-6:

Interleukin-6

LDH:

Lactate dehydrogenase

LncRNA:

Long non-coding RNA

MALAT1:

Metastasis-associated lung adenocarcinoma transcript 1

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

OGD:

Oxygen and glucose deprivation

PBS:

Phosphate-buffered saline

PGC-1α:

Peroxisome proliferator-activated receptor gamma co-activator 1α

PPARγ:

Peroxisome proliferative activated receptorγ

rBMVEC:

Rat brain microvascular endothelial cell

SD rats:

Sprague–Dawley rats

tMCAO:

Transient middle cerebral artery occlusion

TNF-α:

Tumor necrosis factor-α

TTC:

2,3,5-Triphenyltetrazolium chloride

ZO-1:

Zonula occludens-1

References

  • Akinrinmade O, Omoruyi S, Dietrich D, Ekpo O (2017) Long-term consumption of fermented rooibos herbal tea offers neuroprotection against ischemic brain injury in rats. Acta Neurobiol Exp 77(1):94–105

    Google Scholar 

  • Belayev L, Hong SH, Menghani H, Marcell SJ, Obenaus A, Freitas RS, Khoutorova L, Balaszczuk V, Jun B, Oriá RB, Bazan NG (2018) Docosanoids promote neurogenesis and angiogenesis, blood-brain barrier integrity, penumbra protection, and neurobehavioral recovery after experimental ischemic stroke. Mol Neurobiol 55(8):7090-7106

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chang S, Ruan WC, Xu YZ, Wang YJ, Pang J, Zhang LY et al (2017) The natural product 4,10-aromadendranediol induces neuritogenesis in neuronal cells in vitro through activation of the ERK pathway. Acta Pharmacol Sin 38(1):29–40

    Article  CAS  PubMed  Google Scholar 

  • Cheng Y, Zhang HT, Sun L, Guo S, Ouyang S, Zhang Y et al (2006) Involvement of cell adhesion molecules in polydatin protection of brain tissues from ischemia-reperfusion injury. Brain Res 1110(1):193–200

    Article  CAS  PubMed  Google Scholar 

  • Chuang YF, Yang HY, Ko TL, Hsu YF, Sheu JR, Ou G et al (2014) Valproic acid suppresses lipopolysaccharide-induced cyclooxygenase-2 expression via MKP-1 in murine brain microvascular endothelial cells. Biochem Pharmacol 88(3):372–383

    Article  CAS  PubMed  Google Scholar 

  • del Zoppo GJ (2006) Stroke and neurovascular protection. N Engl J Med 354(6):553–555

    Article  PubMed  Google Scholar 

  • del Zoppo GJ (2009) Inflammation and the neurovascular unit in the setting of focal cerebral ischemia. Neuroscience 158(3):972–982

    Article  PubMed  CAS  Google Scholar 

  • Doyle KP, Simon RP, Stenzelpoore MP (2008) Mechanisms of ischemic brain damage. Neuropharmacology 55(3):310–318

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fisher M (2008) Injuries to the vascular endothelium: vascular wall and endothelial dysfunction. Rev Neurol Dis 5(Suppl 1):S4–S11

    PubMed  Google Scholar 

  • Gao Y, Chen T, Lei X, Li Y, Dai X, Cao Y et al (2016) Neuroprotective effects of polydatin against mitochondrial-dependent apoptosis in the rat cerebral cortex following ischemia/reperfusion injury. Mol Med Rep 14(6):5481–5488

  • Guo W, Liu W, Chen Z, Gu Y, Peng S, Shen L et al (2017) Tyrosine phosphatase SHP2 negatively regulates NLRP3 inflammasome activation via ANT1-dependent mitochondrial homeostasis. Nat Commun 8(1):2168

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Hawkins BT, Davis TP (2005) The blood–brain barrier/neurovascular unit in health and disease. Pharmacol Rev 57(2):173–185

    Article  CAS  PubMed  Google Scholar 

  • Heward JA, Lindsay MA (2014) Long non-coding RNAs in the regulation of the immune response. Trends Immunol 35(9):408–419

    Article  CAS  PubMed  Google Scholar 

  • Huang J, Upadhyay UM, Tamargo RJ (2006) Inflammation in stroke and focal cerebral ischemia. Surg Neurol 66(3):232–245

    Article  PubMed  Google Scholar 

  • Huang K, Chen C, Hao J, Huang J, Wang S, Liu P et al (2015) Polydatin promotes Nrf2-ARE anti-oxidative pathway through activating Sirt1 to resist AGEs-induced upregulation of fibronetin and transforming growth factor-β1 in rat glomerular messangial cells. Mol Cell Endocrinol 399:178–189

    Article  CAS  PubMed  Google Scholar 

  • Jinmo K, Miyeon K, Kanghyun L, Sangkwan M, Jamakattel-Pandit N, Hoyoung C et al (2010) Key compound groups for the neuroprotective effect of roots of Polygonum cuspidatum on transient middle cerebral artery occlusion in Sprague-Dawley rats. Nat Prod Res 24(13):1214–1226

    Article  CAS  Google Scholar 

  • Kenneth J, Thomas D (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–∆∆CT method. Methods A 25(4):402–408

    Article  CAS  Google Scholar 

  • Kernan WN, Viscoli CM, Furie KL, Young LH, Inzucchi SE, Gorman M et al (2016) Pioglitazone after ischemic stroke or transient ischemic attack. N Engl J Med 64(1):260

    Google Scholar 

  • Lee G, Elwood F, Mcnally J, Weiszmann J, Lindstrom M, Amaral K et al (2002) T0070907, a selective ligand for peroxisome proliferator-activated receptor γ, functions as an antagonist of biochemical and cellular activities. J Biol Chem 277(22):19649–19657

    Article  CAS  PubMed  Google Scholar 

  • Lisa ML, Derek JP, Randy KB, Jeff EC, Jon LC, Thomas GC et al (2002) Functional consequences of cysteine modification in the ligand binding sites of peroxisome proliferator activated receptors by GW9662. Biochemistry 41(21):6640–6650

    Article  CAS  Google Scholar 

  • Liu J, Wang LN (2017) Peroxisome proliferator-activated receptor gamma agonists for preventing recurrent stroke and other vascular events in people with stroke or transient ischaemic attack. Cochrane Database Syst Rev 12:CD010693

    PubMed  Google Scholar 

  • Liu JY, Yao J, Li XM, Song YC, Wang XQ, Li YJ et al (2014) Pathogenic role of lncRNA-MALAT1 in endothelial cell dysfunction in diabetes mellitus. Cell Death Dis 5:e1506

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Longa EZ, Weinstein PR, Carlson S, Cummins R (1989) Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20(1):84–91

    Article  CAS  PubMed  Google Scholar 

  • Lou T, Jiang W, Xu D, Chen T, Fu Y (2015) Inhibitory effects of polydatin on lipopolysaccharide-stimulated RAW 264.7 cells. Inflammation 38(3):1213–1220

    Article  CAS  PubMed  Google Scholar 

  • Manea SA, Todirita A, Manea A (2013) High glucose-induced increased expression of endothelin-1 in human endothelial cells is mediated by activated CCAAT/enhancer-binding proteins. PLoS ONE 8(12):e84170

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Medhi B, Aggarwal R, Chakrabarti A (2010) Neuroprotective effect of pioglitazone on acute phase changes induced by partial global cerebral ischemia in mice. Indian J Exp Biol 48(8):793–799

    CAS  PubMed  Google Scholar 

  • Michalik KM, You X, Manavski Y, Doddaballapur A, Zörnig M, Braun T et al (2014) Long noncoding RNA MALAT1 regulates endothelial cell function and vessel growth. Circ Res 114(9):1389–1397

    Article  CAS  PubMed  Google Scholar 

  • Puthanveetil P, Chen S, Feng B, Gautam A, Chakrabarti S (2015) Long non-coding RNA malat1 regulates hyperglycaemia induced inflammatory process in the endothelial cells. J Cell Mol Med 19(6):1418–1425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Qureshi IA, Mehler MF (2012) Emerging roles of non-coding RNAs in brain evolution, development, plasticity and disease. Nat Rev Neurosci 13(8):528–541

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ruan W, Zhao F, Zhao S, Zhang L, Shi L, Pang T (2018) Knockdown of long noncoding RNA MEG3 impairs VEGF-stimulated endothelial sprouting angiogenesis via, modulating VEGFR2 expression in human umbilical vein endothelial cells. Gene 649:32–39

    Article  CAS  PubMed  Google Scholar 

  • Ruetzler CA, Furuya K, Takeda H, Hallenbeck JM (2001) Brain vessels normally undergo cyclic activation and inactivation: evidence from tumor necrosis factor-alpha, heme oxygenase-1, and manganese superoxide dismutase immunostaining of vessels and perivascular brain cells. J Cereb Blood Flow Metab 21(3):244–252

    Article  CAS  PubMed  Google Scholar 

  • Sandoval KE, Witt KA (2008) Blood–brain barrier tight junction permeability and ischemic stroke. Neurobiol Dis 32(2):200–219

    Article  CAS  PubMed  Google Scholar 

  • Schellinger PD, Kaste M, Hacke W (2004) An update on thrombolytic therapy for acute stroke. Curr Opin Neurol 17(1):69–77

    Article  CAS  PubMed  Google Scholar 

  • Sharabi K, Hua L, Tavares CDJ, Dominy JE, Camporez JP, Perry RJ et al (2017) Selective chemical inhibition of PGC-1α gluconeogenic activity ameliorates type 2 diabetes. Cell 169(1):148–160

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shi Y, Jiang X, Zhang L, Pu H, Hu X, Zhang W et al (2017) Endothelium-targeted overexpression of heat shock protein 27 ameliorates blood–brain barrier disruption after ischemic brain injury. Proc Natl Acad Sci USA 114(7):E1243–E1252

    Article  CAS  PubMed  Google Scholar 

  • Singh S, Simpson RL, Bennett RG (2015) Relaxin activates peroxisome proliferator-activated receptor γ (PPARγ) through a pathway involving PPARγ coactivator 1α (PGC1α). J Biol Chem 290(2):950–959

    Article  CAS  PubMed  Google Scholar 

  • Su WC, Kang BY, Kim SH, Pak YK, Cho D, Trinchieri G et al (2000) Oxidized low density lipoprotein inhibits interleukin-12 production in lipopolysaccharide-activated mouse macrophages via direct interactions between peroxisome proliferator-activated receptor-γ and nuclear factor-κB. J Biol Chem 275(42):32681–32687

    Article  Google Scholar 

  • Valluri S, Rupam G, Srividya S, George C (2017) Long non-coding rnas regulating immunity in insects. Noncoding RNA 3(1):14

    Google Scholar 

  • Wang Q, Tang XN, Yenari MA (2007) The inflammatory response in stroke. J Neuroimmunol 184(1–2):53–68

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Li C, Chen T, Fang Y, Shi X, Pang T et al (2016) Nafamostat mesilate protects against acute cerebral ischemia via blood–brain barrier protection. Neuropharmacology 105:398–410

    Article  CAS  PubMed  Google Scholar 

  • Wang W, Jiang B, Sun H, Ru X, Sun D, Wang L et al (2017) Prevalence, incidence and mortality of stroke in China: results from a nationwide population-based survey of 480,687 adults. Circulation 135(8):759–771

    Article  PubMed  Google Scholar 

  • Wang Y, Ruan W, Mi J, Xu J, Wang H, Cao Z et al (2018a) Balasubramide derivative 3C modulates microglia activation via CaMKKβ-dependent AMPK/PGC-1α pathway in neuroinflammatory conditions. Brain Behav Immun 67:101–117

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Huang Y, Xu Y, Ruan W, Wang H, Zhang Y et al (2018b) A Dual AMPK/Nrf2 activator reduces brain inflammation after stroke by enhancing microglia M2 polarization. Antioxid Redox Signal 28(2):141–163

    Article  CAS  PubMed  Google Scholar 

  • Wu JS, Kao MH, Tsai HD, Cheung WM, Chen JJ, Ong WY et al (2018) Clinacanthus nutans mitigates neuronal apoptosis and ischemic brain damage through augmenting the C/EBPβ-driven PPAR-γ transcription. Mol Neurobiol 55(7):5425–5438

    Article  CAS  PubMed  Google Scholar 

  • Xie F, Li BX, Kassenbrock A, Xue C, Wang X, Qian DZ et al (2015) Identification of a potent inhibitor of CREB-mediated gene transcription with efficacious in vivo anticancer activity. J Med Chem 58(12):5075–5087

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu Y, Xu Y, Wang Y, Wang Y, He L, Jiang Z et al (2015) Telmisartan prevention of LPS-induced microglia activation involves M2 microglia polarization via CaMKKβ-dependent AMPK activation. Brain Behav Immun 50:298–313

    Article  CAS  PubMed  Google Scholar 

  • Xu M, Yang X, Zeng Q, He H, Lu P, Huang G (2017) Birc5 is a novel target of peroxisome proliferator-activated receptor γ in brain microvascular endothelium cells during cerebral ischemia. Mol Med Rep 16(6):8882–8890

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xue H, Qiao Y, Ni P, Wang J, Chen C, Huang G (2011) A CRE that binds CREB and contributes to PKA-dependent regulation of the proximal promoter of human RAB25 gene. Int J Biochem Cell Biol 43(3):348–357

    Article  CAS  PubMed  Google Scholar 

  • Yao J, Wang XQ, Li YJ, Shan K, Yang H, Wang YN et al (2016) Long non-coding RNA MALAT1 regulates retinal neurodegeneration through CREB signaling. EMBO Mol Med 8(4):346–362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ye J, Piao H, Jiang J, Jin G, Zheng M, Yang J et al (2017) Polydatin inhibits mast cell-mediated allergic inflammation by targeting PI3K/Akt, MAPK, NF-κB and Nrf2/HO-1 pathways. Sci Rep 7(1):11895

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Yin KJ, Hamblin M, Chen YE (2014) Non-coding RNAs in cerebral endothelial pathophysiology: emerging roles in stroke. Neurochem Int 77:9–16

    Article  CAS  PubMed  Google Scholar 

  • Zhang JH, Badaut J, Tang J, Obenaus A, Hartman R, Pearce WJ (2012) The vascular neural network-a new paradigm in stroke pathophysiology. Nat Rev Neurol 8(12):711–716

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang J, Yuan L, Zhang X, Hamblin MH, Zhu T, Meng F et al (2016) Altered long non-coding RNA transcriptomic profiles in brain microvascular endothelium after cerebral ischemia. Exp Neurol 277:162–170

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Tang X, Liu K, Hamblin MH, Yin KJ (2017) Long non-coding RNA Malat1 regulates cerebrovascular pathologies in ischemic stroke. J Neurosci 37(7):1797–1806

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao G, Jiang K, Wu H, Qiu C, Deng G, Peng X (2017) Polydatin reduces Staphylococcus aureus lipoteichoic acid-induced injury by attenuating reactive oxygen species generation and TLR2-NFkB signalling. J Cell Mol Med 21(11):2796–2808

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zilleßen P, Celner J, Kretschmann A, Pfeifer A, Racké K, Mayer P et al (2016) Metabolic role of dipeptidyl peptidase 4 (DPP4) in primary human (pre)adipocytes. Sci Rep 6:23074

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (81570236, 81870360, 81402385, 81571134, 81773995, 81320108029), the Natural Science Foundation of Jiangsu Province (BK20160032), “Double First-Class” University Project (CPU2018GY06, CPU2018GY20), the Six Talent Peaks Project of Jiangsu Province (T.P.), Shanghai Key Laboratory of Psychotic Disorders (13dz2260500), the Postgraduate Research & Practice Innovation Program of Jiangsu Province. We would like to acknowledge Dr. Xiujun Li of the Affiliated Drum Tower Hospital of Nanjing University Medical School, for providing pEGFP-C1-MALAT1 and control plasmids; Dr. Shanshan Guo of Ningxia Medical University for providing the C/EBPβ overexpression plasmid pcDNA3.1-C/EBPβ; Prof. Qin Jiang of Nanjing Medical University for generously providing us with the rat MALAT1 primer sequence. We also gratefully acknowledge the excellent technical assistance of Haojie Wang, Chenglong Gao, Tailin He and Xin Guan in the animal experiments.

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Author notes

  1. Wenchen Ruan and Jingwei Li have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Drug Screening, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, China Pharmaceutical University, #24 Tong Jia Xiang Street, Nanjing, 210009, People’s Republic of China

    Wenchen Ruan, Yazhou Xu, Yunjie Wang, Hulin Jiang, Luyong Zhang & Tao Pang

  2. College of Basic Medical Sciences, Dalian Medical University, No. 9 West Section Lvshun South Road, Dalian, 116044, People’s Republic of China

    Wenchen Ruan, Feng Zhao, Xu Yang & Lei Shi

  3. Department of Neurology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, 210008, People’s Republic of China

    Jingwei Li

  4. Center for Drug Screening and Pharmacodynamics Evaluation, School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou, 510006, People’s Republic of China

    Luyong Zhang

  5. Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing, 210009, People’s Republic of China

    Luyong Zhang & Tao Pang

  6. Department of Pharmacology and Physiology, Georgetown University Medical Center, Washington, DC, 20057, USA

    Juan M. Saavedra & Tao Pang

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  1. Wenchen Ruan
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  2. Jingwei Li
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  3. Yazhou Xu
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  4. Yunjie Wang
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  10. Lei Shi
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Contributions

All authors listed contributed immensely to this study. WR and JL performed the experiments and wrote the paper. YX, YW, FZ, and XY performed the animal experiments and analyzed the data. HJ, LZ, JMS, LS, TP, as experts in molecular pharmacology provided technical supports and designed the research.

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Correspondence to Lei Shi or Tao Pang.

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Ruan, W., Li, J., Xu, Y. et al. MALAT1 Up-Regulator Polydatin Protects Brain Microvascular Integrity and Ameliorates Stroke Through C/EBPβ/MALAT1/CREB/PGC-1α/PPARγ Pathway. Cell Mol Neurobiol 39, 265–286 (2019). https://doi.org/10.1007/s10571-018-00646-4

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