Bilirubin-induced calcium overload in synaptosomes isolated from brainstem neurons of rats
10.3969/j.issn.1674-8115.2017.11.002
- VernacularTitle:胆红素导致SD大鼠脑干神经元突触小体Ca2+过载
- Author:
ping Dan LI
1
;
Ke LAI
;
ping Ji WANG
;
bo Hai SHI
Author Information
1. 上海交通大学附属第六人民医院耳鼻咽喉头颈外科
- Keywords:
bilirubin;
synaptosome;
intracellular calcium;
glycoursodeoxycholic acid;
fluorescent probe
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2017;37(11):1458-1462
- CountryChina
- Language:Chinese
-
Abstract:
Objective· To observe real-time changes of calcium concentration ([Ca2+]i) exposure to bilirubin in synaptosomes isolated from brainstem nucleus of rats. Methods · Forty P7-14 SD rats were randomly assigned to three groups: control group, bilirubin group (with levels of 0.1, 1 and 10 μmol/L) and bulirubin plus glycoursodeoxycholic acid (GUDCA) group. The synaptosomes were purified from brainstem nucleus by sucrose density gradient centrifugation. After loading OG-BAPTA in synaptosomes, two dimensional image of intracellular calcium and analysis of fluorescence intensity were achieved by Confocal laser scanning microscopy. Results · Synaptosomes with well biological activity were obtained from brainstem of the SD rats. In the control group, a progressive increase in fluorescent intensity of [Ca2+]I was detected. In the bilirubin group, acuter increases in fluorescent intensity were observed in all levels of bilirubin, with a manner of both concentration and time-dependent (P<0.05). Fluorescent intensity of [Ca2+]I was reduced in the present of GUDCA, which was not significant compared with the control group (P=0.656). However, GUDCA could abate the increase of fluorescent intensity of [Ca2+]I induced by bilirubin exposure, of which showing significant decrease in 10 μmol/L bilirubin exposure (P=0.000). Conclusion · Bilirubin could induce calcium overload in synaptosomes. GUDCA could abate bilirubin-induced calcium overload in synaptosomes, possibly explaining its protection effect of neurons from bilirubin neurotoxicity.
