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Autophagy inhibits high glucose induced cardiac microvascular endothelial cells apoptosis by mTOR signal pathway

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Abstract

Cardiac microvascular endothelial cells (CMECs) dysfunction is an important pathophysiological event in the cardiovascular complications induced by diabetes. However, the underlying mechanism is not fully clarified. Autophagy is involved in programmed cell death. Here we investigated the potential role of autophagy on the CMECs injury induced by high glucose. CMECs were cultured in normal or high glucose medium for 6, 12 and 24 h respectively. The autophagy of CMECs was measured by green fluorescence protein (GFP)-LC3 plasmid transfection. Moreover, the apoptosis of CMEC was determined by flow cytometry. Furthermore, 3-Methyladenine (3MA), ATG7 siRNA and rapamycin were administrated to regulate the autophagy state. Moreover, Western blotting assay was performed to measure the expressions of Akt, mTOR, LC3 and p62. High glucose stress decreased the autophagy, whereas increased the apoptosis in CMECs time dependently. Meanwhile, high glucose stress activated the Akt/mTOR signal pathway. Furthermore, autophagy inhibitor, 3-MA and ATG7 siRNA impaired the autophagy and increased the apoptosis in CMECs induced by high glucose stress. Conversely, rapamycin up-regulated the autophagy and decreased the apoptosis in CMECs under high glucose condition. Our data provide evidence that high glucose directly inhibits autophagy, as a beneficial adaptive response to protect CMECs against apoptosis. Furthermore, the autophagy was mediated, at least in part, by mTOR signaling.

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Acknowledgements

This work was supported by National Nature Science Foundation of China (No. 81400274).

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    Authors and Affiliations

    1. Department of Cardiology, The General Hospital of the PLA Rocket Force, Beijing, 100088, China

      Zheng Zhang, Zhitao Jin, Liping Ding, Wei Jiang, Junke Yang, Zhimin Sun & Taohong Hu

    2. Department of Cardiology, The Seventh People’s Hospital of Zhengzhou, Zhengzhou, 450016, Henan, China

      Shenwei Zhang & Chunguang Qiu

    3. Department of Nuclear Medicine, The Affiliate Hospital of Military Medicine Academy, Beijing, 100071, China

      Yong Wang

    4. School of Basic Medical Sciences, Taishan Medical University, Taian, 271000, Shandong, China

      Ming Yang

    5. Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai, 201306, China

      Ming Yang

    6. Department of Outpatient, The General Hospital of the PLA Rocket Force, Beijing, 100088, China

      Ning Zhang

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    1. Zheng Zhang
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    Correspondence to Chunguang Qiu or Taohong Hu.

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    All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

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    Zheng Zhang, Shenwei Zhang, Yong Wang, and Ming Yang have contributed equally to this work.

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    10495_2017_1398_MOESM1_ESM.tif

    Supplementary material 1 HG induced apoptosis via caspase way. A: Representative TUNEL staining imaging of CMECs in each group. B: Quantification of the apoptotic CMECs (n=3, *p < 0.05). C: Representative Western blots of cleaved caspase-3 in each group. D: Quantitative analysis of the expressions of cleaved caspase-3/β actin. (TIF 1619 KB)

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    Zhang, Z., Zhang, S., Wang, Y. et al. Autophagy inhibits high glucose induced cardiac microvascular endothelial cells apoptosis by mTOR signal pathway. Apoptosis 22, 1510–1523 (2017). https://doi.org/10.1007/s10495-017-1398-7

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