Effects of different ratios of medicine dosage for isoflurane and propofol on GABAAreceptor α1sub-unit proteostasis during hypoxia injury to hippocampal neurons of rats
10.3760∕cma.j.issn.0254-1416.2017.11.015
- VernacularTitle:异氟醚∕丙泊酚不同配伍对大鼠海马神经元缺氧损伤时GABAA受体α1亚基稳态的影响
- Author:
Tang LI
1
;
Haiyun WANG
;
Di GUO
;
Xinyue WANG
;
Guolin WANG
Author Information
1. 300170,天津医科大学三中心临床学院 天津市第三中心医院麻醉科
- Keywords:
Propofol;
Isoflurane;
Receptors,GABA-A;
Hypoxia-ischemia,brain
- From:Chinese Journal of Anesthesiology
2017;37(11):1336-1341
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the effects of different ratios of medicine dosage for isoflurane and propofol on GABAAreceptor(GABAAR)α1subunit proteostasis during hypoxia injury to hippocampal neurons of rats. Methods The hippocampal neurons isolated from fetal rats obtained from Wistar rats were primarily cultured and divided into 6 groups(n=60 each)using a random number table: control group (group C), hypoxia group(group H), isoflurane group(group I), propofol group(group P)and dif-ferent ratios of medicine dosage for isoflurane and propofol groups(group IP1and group IP2). The cells were subjected to hypoxia for 6 h in group H. Cells were incubated for 3 h with 1.9 % isoflurane and with 22.4 μmol∕L propofol after being subjected to hypoxia for 6 h in I and P groups, respectively. Cells were incubated for 3 h with 1.0% isoflurane and 6.7 μmol∕L propofol and with 1.4% isoflurane and 3.4 μmol∕L propofol after being subjected to hypoxia for 6 h in IP1and IP2groups, respectively. Then the culture medi-um was replaced with plain culture medium. At 24 h of incubation, the cells were collected for measure-ment of cell viability by CCK-8 assay, GABAAR α1mRNA expression(by quantitative polymerase chain reaction), GABAAR α1expression in the cytomembrane(by Western blot), level of GABAAR α1subunit endoplasmic reticulum-associated degradation(ERAD)(by immunoprecipitation and Western blot)and CCAAT∕enhancer-binding protein homologous protein(CHOP)expression(by immunofluorescence). Re-sults Compared with group C, the cell viability was significantly decreased, the expression of GABAAR α1mRNA and GABAAR α1in cytomembrane was down-regulated, the expression of CHOP was up-regula-ted, and the level of GABAAR α1subunit ERAD was increased in the other five groups(P<0.05). Com-pared with group H, the cell viability was significantly decreased, the expression of GABAAR α1mRNA and GABAAR α1in cytomembrane was down-regulated, the expression of CHOP was up-regulated, and the level of GABAAR α1subunit ERAD was increased in I, P and IP2groups(P <0.05), and no significant change was found in the parameters mentioned above in group IP1(P<0.05). Compared with group I or group P, the cell viability was significantly increased, the expression of GABAAR α1mRNA and GABAAR α1 in cytomembrane was up-regulated, the expression of CHOP was down-regulated, and the level of GABAAR α1subunit ERAD was decreased in IP1and IP2groups(P<0.05). Compared with group IP1, the cell viability was significantly decreased, the expression of GABAAR α1mRNA and GABAAR α1in cy-tomembrane was down-regulated, the expression of CHOP was up-regulated, and the level of GABAAR α1 subunit ERAD was increased in group IP2(P<0.05). Conclusion Combination of 1.0% isoflurane and 6.7 μmol∕L propofol does not aggravate hypoxia-induced destruction of GABAAR α1subunit proteostasis in hippocampal neurons of rats.
