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Cellular and Molecular Neurobiology

, Volume 39, Issue 2, pp 265–286 | Cite as

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

  • Wenchen Ruan
  • Jingwei Li
  • Yazhou Xu
  • Yunjie Wang
  • Feng Zhao
  • Xu Yang
  • Hulin Jiang
  • Luyong Zhang
  • Juan M. Saavedra
  • Lei ShiEmail author
  • Tao PangEmail author
Original Research

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.

Keywords

MALAT1 Ischemic stroke Cerebrovascular endothelial cells Polydatin 

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

Notes

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.

Author 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.

Compliance with Ethical Standards

Conflict of interest

The authors declare that there are no competing interests.

Supplementary material

10571_2018_646_MOESM1_ESM.doc (20 mb)
Supplementary material 1 (DOC 20441 KB)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 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 BiopharmaceuticalsChina Pharmaceutical UniversityNanjingPeople’s Republic of China
  2. 2.College of Basic Medical SciencesDalian Medical UniversityDalianPeople’s Republic of China
  3. 3.Department of NeurologyThe Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjingPeople’s Republic of China
  4. 4.Center for Drug Screening and Pharmacodynamics Evaluation, School of PharmacyGuangdong Pharmaceutical UniversityGuangzhouPeople’s Republic of China
  5. 5.Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University)Ministry of EducationNanjingPeople’s Republic of China
  6. 6.Department of Pharmacology and PhysiologyGeorgetown University Medical CenterWashingtonUSA

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