Telomerase reverse transcriptase promotes angiogenesis in neonatal rats after hypoxic-ischemic brain damage

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Biochemistry, Biophysics and Molecular Biology

Main article text

 

Introduction

Materials & Methods

Experimental animals

Brain microvascular endothelial cells (BMECs) cultured in vitro

Construction of vectors and lentiviral infection of BMECs

Oxygen glucose deprivation treatment

Infection with adenovirus and hypoxia-ischemia treatment in vivo

Quantitative real-time PCR

Cell counting and kit-8 staining

Western blot

Immunofluorescence staining

Immunohistochemical assay

Microvessel formation assay

Evaluation of blood–brain barrier permeability

Statistical analysis

Results

TERT overexpression increases the proliferation rate of BMECs after OGD

TERT overexpression increases the formation of microvessels in the brain after HIBD both in vivo and in vitro

TERT overexpression improves BBB integrity after HIBD

TERT increases Notch-1 levels in BMECs following oxygen glucose deprivation

Discussion

Conclusions

Supplemental Information

Raw data

Immunofluorescence staining pictures, immunohistochemical staining pictures and uncropped gel photo, and the statistical analysis.

DOI: 10.7717/peerj.14220/supp-1

ARRIVE 2.0 Checklist

DOI: 10.7717/peerj.14220/supp-2

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests.

Author Contributions

Jiao Li performed the experiments, prepared figures and/or tables, and approved the final draft.

Yi Feng performed the experiments, prepared figures and/or tables, and approved the final draft.

Jing Zhao analyzed the data, prepared figures and/or tables, and approved the final draft.

Zhi Fang conceived and designed the experiments, authored or reviewed drafts of the article, and approved the final draft.

Haiting Liu conceived and designed the experiments, authored or reviewed drafts of the article, and approved the final draft.

Animal Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

Under the approval of the Sichuan University Committee of Animal Research, neonatal Sprague Dawley (SD) rats (30 rats, 15 males and 15 females, 5 days old, weight of 15–20 g) were provided by the Animal Center of Sichuan Province. Animal protocols were performed in compliance with the protocols approved by the Animal Ethics Committee of West China Second University Hospital, Sichuan University (approval number, 2021034). All animal experiments in this study complied with the Animal Research: Reporting of In Vivo Experiments Guidelines and were carried out in accordance with the recent guidelines. Furthermore, the work in this study followed the EC Directive 86/609/EEC for animal experiments and the Uniform Requirements for manuscripts submitted to Biomedical Journals.

Data Availability

The following information was supplied regarding data availability:

The raw data is available in the Supplemental Files.

Funding

Jiao Li received support from the National Natural Science Foundation of China (grant #81501302). Zhi Fang received support from The Grant from the Science and Technology Bureau of Sichuan Province (grant #2021YFS0247; #2022YFS0360) and the Foundation of Health and Family Planning Commission of Sichuan Province (CN) (grant #17ZD028). The New Bud foundation of West China Second University Hospital, Sichuan University provided support through grant #Kx105, to Jiao Li and grant #Kx097, to Haiting Liu. The National Natural Science Foundation of China contributed to the design of the study (grant #81501302, to Jiao Li). The Foundation of Health and Family Planning Commission of Sichuan Province (CN) (grant #17ZD028, to Zhi Fang) and the New Bud foundation of West China Second University Hospital, Sichuan University (grant #Kx105, to Jiao Li; grant #Kx097, to Haiting Liu), contributed to the collection, analysis and interpretation of data, the writing of the report, and the decision to submit the article for publication.

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