Skip to main content
main-content

01.12.2012 | Study protocol | Ausgabe 1/2012 Open Access

Cardiovascular Diabetology 1/2012

Detecting DNA synthesis of neointimal formation after catheter balloon injury in GK and in Wistar rats: using 5-ethynyl-2'-deoxyuridine

Zeitschrift:
Cardiovascular Diabetology > Ausgabe 1/2012
Autoren:
Jingsheng Guo, Dongye Li, Shiru Bai, Tongda Xu, Zhongmin Zhou, Yanbin Zhang
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2840-11-150) contains supplementary material, which is available to authorized users.

Competing interests

The authors declare that they have no competing interests.

Authors’ contributions

JG contributed to the in vivo experiments, participated in the pathological evaluation, interpretation, and acquisition of data, performed statistical analysis, drafted portions of the manuscript, and critically revised the manuscript before final approval. DL participated in the overall design, analysis and interpretation of the data, as well as the writing and presentation of this work. SB participated in the animal model experiments, the data collection and interpretation, performed statistical analysis, drafted portions of the manuscript, and critically revised the manuscript before final approval. TX contributed to the establishment of the animal model as well as the data collection and interpretation. ZZ participated in the model design and the overall conduct of the study. YZ contributed to the study design and data interpretation. All authors read and approved the final manuscript.

Abstract

Background

Neointimal formation plays an important role in the pathogenesis of coronary restenosis after percutaneous coronary intervention (PCI), especially in patients with diabetes mellitus. Recently, some studies have shown that 5-ethynyl-2'-deoxyuridine (EdU) incorporation can serve as a novel alternative to the 5-bromo-2'-deoxyuridine (BrdU) antibody detection method for detection of DNA synthesis in regenerating avian cochlea, chick embryo and the adult nervous system. However, few studies have been performed to assess the suitability of EdU for detecting DNA synthesis in vascular neointima.

Methods

The carotid artery balloon injury model was established in Goto-Kakizaki (GK) and Wistar rats. A Cell-LightTM EdU Kit was used to detect EdU-labeled cell nuclei of common carotid arteries at day 7 after catheter balloon injury. Different methods of injecting EdU were tested. The protein levels of proliferating cell nuclear antigen (PCNA) and p-Akt (Ser473), as well as the mRNA levels of PCNA were evaluated by Western blotting and quantitative real-time PCR (qRT-PCR), respectively. Immunohistochemical staining was also employed to visualize PCNA-positive cells.

Results

At day 7 after catheter balloon injury, far more EdU-positive and PCNA-positive cells were observed in GK rats. When comparing groups that received different EdU doses, it was found that the percentage of EdU-positive cells at a dose of 100 mg/kg body weight was than at doses of 25 mg/kg and 50 mg/kg. The number of positive cells was significantly higher in the repeated injection group compared to the single injection group. Further, after balloon injury DNA synthesis in GK rats was more notable than in Wistar rats. Neointimal formation in GK rats was more obvious than in Wistar rats. The protein levels of PCNA and p-Akt (Ser473) and the mRNA levels of PCNA were increased in injured rats as compared to uninjured rats, and were significantly higher in GK rats than in Wistar rats.

Conclusion

By intraperitoneal injections of EdU at a dose of 100 mg/kg three times, EdU incorporation can detect carotid arterial DNA synthesis caused by neointimal formation in GK rats and Wistar rats at day 7 after balloon injury by the EdU click reaction quickly and effectively. Moreover, more obvious DNA synthesis in the vascular neointima could be observed in GK rats than in Wistar rats.
Zusatzmaterial
Authors’ original file for figure 1
12933_2012_574_MOESM1_ESM.tiff
Authors’ original file for figure 2
12933_2012_574_MOESM2_ESM.tiff
Authors’ original file for figure 3
12933_2012_574_MOESM3_ESM.tiff
Authors’ original file for figure 4
12933_2012_574_MOESM4_ESM.tiff
Authors’ original file for figure 5
12933_2012_574_MOESM5_ESM.tiff
Authors’ original file for figure 6
12933_2012_574_MOESM6_ESM.tiff
Literatur
Über diesen Artikel

Weitere Artikel der Ausgabe 1/2012

Cardiovascular Diabetology 1/2012 Zur Ausgabe

Neu im Fachgebiet Innere Medizin

Meistgelesene Bücher aus der Inneren Medizin

2017 | Buch

Rheumatologie aus der Praxis

Entzündliche Gelenkerkrankungen – mit Fallbeispielen

Dieses Fachbuch macht mit den wichtigsten chronisch entzündlichen Gelenk- und Wirbelsäulenerkrankungen vertraut. Anhand von über 40 instruktiven Fallbeispielen werden anschaulich diagnostisches Vorgehen, therapeutisches Ansprechen und der Verlauf …

Herausgeber:
Rudolf Puchner

2016 | Buch

Ambulant erworbene Pneumonie

Was, wann, warum – Dieses Buch bietet differenzierte Diagnostik und Therapie der ambulant erworbenen Pneumonie zur sofortigen sicheren Anwendung. Entsprechend der neuesten Studien und Leitlinien aller wichtigen Fachgesellschaften.

Herausgeber:
Santiago Ewig

Mail Icon II Newsletter

Bestellen Sie unseren kostenlosen Newsletter Update Innere Medizin und bleiben Sie gut informiert – ganz bequem per eMail.

© Springer Medizin 

Bildnachweise