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
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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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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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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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DOI: https://doi.org/10.1007/s10571-018-00646-4