Role of PI3K/Akt in endothelial progenitor cells differentiationin vitro

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Abstract

BACKGROUND:

The molecule mechanism underlying endothelial progenitor cells (EPCs) differentiation into mature endothelial cells remains poorly understood. It requires for the regulation of the expression of various genes inside cells by various signal pathways, PI3K/Akt maybe included.

OBJECTIVE:

To investigate the role of PI3K/Akt in EPCs differentiation.

DESIGN, TIME AND SETTING:

A group control in vitro experiment was performed at the Pathology Laboratory in Shengjing Hospital of China Medical University from September 2007 to March 2008. MATERIALS: Wistar rats aged 4-6 weeks, of either gender.

METHODS:

EPCs were isolated from rat bone marrow with density gradient centrifugation. Then difference-speed adherence screening method was used to obtain the second time adherent cells, which then were inoculated in culture flask. After 5 days of culture, cells were collected to be indentified with laser confocal microscopy. The differentiating EPCs were those that have positive results of both AC133 and vWF stainings.

MAIN OUTCOME MEASURES:

The mRNA and the protein expression of AC133, vWF, PI3K and Akt in EPCs were detected with Western blot and reverse transcription-polymerase chain reaction (RT-PCR) after 0, 3, 7, 10 and 14 days of inoculation respectively.

RESULTS:

According to the RT-PCR and Western blot detection, AC133 showed the strongest expression at day 0, the weaker at day 3 and none at day 7, 10 and 14 (P < 0.05); vWF presented no significant expression change throughout the whole inoculation; the expressions of PI3K and Akt changed in the same direction with AC133, i.e. they became stronger with the increasing culture time, with the strongest at day 0, and the weaker at day 3.

CONCLUSION:

PI3K/Akt signal pathway possibly has its role in the differentiation of EPCs into endothelial cells.

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