Growth enhancement effect of BzATP on primary cultured astrocytes from rat brain.
OBJECTIVE
To explore whether BzATP could promote the growth of primary cultured astrocytes (AS) of rat and its possible mechanism, and whether TGF-β1 was involved in the event.


METHODS
The primary cultured ASs were derived from new born Sprague-Dawley rats. Glial fibrillary acidic protein (GFAP) immunofluorescent staining was used to check the purity of cultured AS. Morphometry was used to detect the changes of AS. The proliferation index of AS was detected by BrdU incorporation assay. Western blot was used to detect the changes of GFAP under different conditions. Changes of TGF-β1 gene transcription were detected by RT-PCR. ELISA was utilized to detect the variation of TGF-β1 protein in the supernate.


RESULTS
The purity of primary cultured AS reached 99%. BzATP promoted the hypertrophy of AS including the elongation of AS processes and the enlargement of cell bodies, BzATP also promoted the expression of GFAP in existence of Ca²⁺, but had no effect on cell proliferation. BzATP increased the transcription of TGF-β1 mRNA and the release of TGF-β1 protein in existence of Ca²⁺. TGF-β1 neutralizing antibody partially inhibited the expression of GFAP induced by BzATP, but had no effect on AS proliferation and cell morphology.


CONCLUSIONS
BzATP enhanced the hypertrophy of primary cultured AS, increased the expression of GFAP partially through TGF-β1. Mechanisms of the enhancement of AS growth induced by BzATP other than TGF-β1 pathway remain to be elucidated.
Liang
Hua-Zheng
HZ
Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Liu
Ying
Y
Ye
Zhu-Rong
ZR