Cell Physiol Biochem 2015;37: DOI: / Published online: September 11, 2015

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1 /15/ $39.50/0 Qi Accepted: et al.: JMJD2A August Inhibition 06, 2015 Attenuates Neointimal Hyperplasia in Diabetic Rats Original Paper This is an Open Access article licensed under the terms of the Creative Commons Attribution- NonCommercial 3.0 Unported license (CC BY-NC) ( applicable to the online version of the article only. Distribution permitted for non-commercial purposes only. Histone Demethylase JMJD2A Inhibition Attenuates Neointimal Hyperplasia in the Carotid Arteries of Balloon-Injured Diabetic Rats via Transcriptional Silencing: Smooth Muscle Cells Hu Qi a Zhang Jing b Wu Xiaolin c Xu Changwu a Hu Xiaorong a Yang Jian b Chen Jing a Jiang Hong a a Department of Cardiology, Renmin Hospital of Wuhan University, Cardiovascular Research Institute of Wuhan University, Wuhan, b Department of Cardiology, The First College of Clinical Medical Sciences, China Three Gorges University, Yichang, Hubei, c Department of Cardiology, Xiangyang Central Hospital, Key Words muscle cell Abstract Background/Aims: Diabetic patients suffer from severe neointimal hyperplasia following angioplasty. The epigenetic abnormalities are increasingly considered to be relevant to the pathogenesis of diabetic cardiovascular complications. But the epigenetic mechanisms linking diabetes and coronary restenosis have not been fully elucidated. In this study, we explored the protective effect and underlying mechanisms of demethylases JMJD2A inhibition in ballooninjury induced neointimal formation in diabetic rats. Methods: JMJD2A inhibition was achieved by the chemical inhibitor 2,4-pyridinedicarboxylic acid (2,4-PDCA) and small interfering RNA (sirna). In vitro In vivo, diabetic rats induced using high-fat diet and low-dose streptozotocin (35mg/kg) underwent carotid artery balloon injury. Morphometric analysis was performed using hematein eosin and immumohistochemical staining. Results: The global JMJD2A was increased in vitro. And JMJDA2A inhibition attenuated Jiang Hong and Chen Jing Department of Cardiology, Renmin Hospital of Wuhan University, Cardiovascular Research Institute of Wuhan University, 238 Jiefang Road, Wuhan , (China) Fax , pyh_hongj@163.com

2 720 neointimal formation in balloon-injured diabetic rats. The underlying mechanisms were relevant to the restoration of H3K9me3 levels at the promoters of MCP-1 and IL-6, and then the suppressed expression of MCP-1 and IL-6. Conclusion: Copyright Introduction 2+ in vitro in vivo Materials and Methods Animals and ethics statement

3 ,4-pyridinedicarboxylic acid (2,4-PDCA) treatment in vitro and in vivo in vitroin vivo In vitro mm). In vivo Small interfering RNA transfection in vitro and in vivo in vitro, in vivo Cell culture and high glucose, fatty acid and insulin treatment 2 th Cell proliferation Cell migration 5 Chromatin immunoprecipitation (ChIP) assay

4 722 using the 2 Diabetic rat model and carotid artery balloon injury Metabolic measurements K Morphometric analysis th Quantitative real-time PCR

5 723 Western blotting Statistical analysis Results JMJD2A protein levels are increased in HG-stimulated VSMCs and balloon-injured carotid arteries of diabetic rats, whereas H3K9me3 levels are decreased In vitro JMJD2A inhibition suppressed HG-induced proliferation, migration via the down-regulated

6 724 Fig. 1. * p # p ## p

7 725 Fig. 2. * p ** p * p ** p # p ## p * p ** p # p ## p # p ## p

8 726 Fig. 3. * p ** p restoration of H3K9me3 at promoters JMJD2A inhibition alleviates neointimal formation in diabetic rats in vivo

9 727 Fig. 4. * p ** p # p ## p * p ** p # p ## p Table 1. p

10 728 Fig. 5. ## p # p ## p * p ** p ## p ** p

11 729 Fig. 6. th in vivo # p ## p * p ** p th in vivo # p ## p * p ** p ## p * p ** p arteries of diabetic rats in vivo

12 730 in vivo in vivo Discussion in vitro in vivo. In vitro in vitro

13 731 in vitroin vivo in vivo in vitro Conclusions Acknowledgements

14 732 Disclosure Statement References

15 733

16 734 In vitro in vitro in vitro

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