The Effect of Therapeutic Mild Hypothermia on Brain Microvascular Endothelial Cells During Ischemia–Reperfusion Injury

  • Yao Chen
  • Lin Wang
  • Yun Zhang
  • Yaxiong Zhou
  • Wei Wei
  • Zhi Wan
Translational Research
  • 121 Downloads

Abstract

Background

To determine the cerebral protective effects of mild hypothermia (MH) on cerebral microcirculation.

Methods

We established ischemia–reperfusion (I/R) injury and MH treatment models with rat brain microvascular endothelial cells (RBMECs) in vitro and examined the apoptotic changes. The cultured RBMECs were randomly divided into the control group, I/R group, and MH group, which was further divided into two subgroups: intra-ischemia hypothermia (IIH) and post-ischemia hypothermia (PIH). Cell morphological changes were assessed using fluorescence microscopy. Apoptotic rates were obtained by flow cytometry. Expressions of caspase-3, Bax, and Bcl-2 were analyzed by Western blot.

Results

I/R injury in vitro induced apoptosis of RBMECs. The apoptotic rates in the control group, I/R group, and MH group were 0.13, 19.04, and 13.13%, respectively (P < 0.01). Compared with the I/R group, the MH group showed a significant decrease in the number of apoptotic cells, mainly in stage I apoptotic cells (P < 0.0083). The caspase-3 and Bax expressions were significantly enhanced (P < 0.05) in RBMECs after I/R injury, while substantial decreases in Bcl-2 expression were noted (P < 0.05). Following MH intervention, the increase in caspase-3 and Bax expression was suppressed (P < 0.05), while Bcl-2 expression significantly increased. The apoptotic rates or protein expressions between the two subgroups were not different significantly (P > 0.05).

Conclusions

These results indicate that MH could inhibit RBMEC apoptosis by preventing pro-apoptotic cells and early apoptotic cells from progressing to intermediate and advanced stages. This may be due to the effect of MH on I/R-induced apoptotic gene expression changes.

Keywords

Cardiopulmonary resuscitation Cerebral microcirculation Brain microvascular endothelial cells Ischemia–reperfusion injury Mild hypothermia Apoptosis Timing of hypothermia 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC: 81201446, https://isis.nsfc.gov.cn). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors declare no conflicts of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  • Yao Chen
    • 1
  • Lin Wang
    • 2
  • Yun Zhang
    • 3
  • Yaxiong Zhou
    • 1
  • Wei Wei
    • 1
  • Zhi Wan
    • 1
  1. 1.Department of Emergency MedicineWest China Hospital, Sichuan UniversityChengduChina
  2. 2.Department of CardiologyChengdu Shangjin Jin Nanfu HospitalChengduChina
  3. 3.Department of EmergencyWuxi People’s Hospital, NanJing Medical UniversityWuxiChina

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