Inhibitive effect of cliary neurotrophic factor on thapsigargin-induced increase of intracellular -[Ca~(2+)-]_i in cultured hippocampal neurons / 第二军医大学学报

ABSTRACT

Objective:To observe the effect of ciliary neurotrophic factor (CNTF) on the increase of hippocampal neurons -[Ca~(2+)-]_i induced by thapsigargin (Tg), a sarco/endoplasmic reticulum Ca~(2+)-ATPase (SERCA2a) inhibitor. Methods: The primarily cultured hippocampal neurons of newborn SD rats(within 24 h of delivery) were prepared after 10-14 d culture. The intracellular -[Ca~(2+)-]_i was measured with Fura-2/AM in real time. The neurons were divided into 3 groups. Group A (n=20), EGTA (3 mmol/L);Group B (n=40), EGTA (3 mmol/L) + Tg (10 ?mol/L);and Group C (n=12), EGTA (3 mmol/L) +Tg (10 ?mol/L) +CNTF (500 U/?l). Results: When the extracellular Ca~(2+) was chelated by enough EGTA, -[Ca~(2+)-]_i was ≤(30.73?7.25) nmol/L; after EGTA+Tg was added, -[Ca~(2+)-]_i significantly increased(-[Ca~(2+)-]_i≥-[91.55?12.24-] nmol/L); and after EGTA+Tg+CNTF was added, -[Ca~(2+)-]_i was markedly reduced compared with that in group B (-[Ca~(2+)-]_i≤-[26.09?7.16-] nmol/L, P