RhoA is involved in thrombin-induced neuron injury in the cortex of fetal rats.
10.3760/cma.j.issn.1671-8925.2012.06.008
- VernacularTitle:RhoA蛋白参与凝血酶对胎鼠大脑皮层神经元的损伤
- Author:
Hai TANG
1
;
Xing-Qi WANG
;
Xia SHEN
;
Hui ZHAO
;
Li GAO
;
Hao CHEN
;
Gui-Yun CUI
Author Information
1. 徐州医学院附属医院
- Keywords:
Intracerebral hemorrhage;
Thrombin;
RhoA;
Cortex neuron
- From:
Chinese Journal of Neuromedicine
2012;11(6):570-574
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the relationship between the activation of RhoA and thrombin-induced neuron injury in the cortex of fetal rats. Methods The neurons from the fetal ratcortex were culturedin vitro for 8 d; and then,they were treated by thrombin at different concentrations (0,1,10,30 and 100 U/mL) for 3 h,and by 30 U/mL thrombm at different times (0,0.5,1,3 and 6 h);Western blotting was used to examine the effects of these different treatments on the activation of RhoA.The neurons were pretreated with Exoenzyme C3,the RhoA inhibitor,for 0.5 h,and incubated with 30 U/ml thrombin for 3 h; and then,Western blotting was employed to examine the activation of Rho A; besides that,the injuries of these neurons with the presence and absence of Exoenzyme C3 were observed by Hoechst33258 staining and CCK-8 assay.Results The activation of RhoA expressing in membrane with the treatment of 30 U/mL thrombin for 3 h was significantly increased as compared with that under the treatment of 0 U/mL (P＜0.05); and the total RhoA showed no significant changes with the treatment of all concentrations. The 3 h site with thrombin (30 U/mL) could more significantly induce R hoA expression as compared with other time sites (P＜0.05),and the total RhoA showed no significant changes under the treatment of all time sites. Pretreatrnent of neurons with Exoenzyme C3 could significantly down-regulate RhoA level as compared with those without pretreatment (P＜0.05); meanwhile,Hoechst33258 staining indicated that the number of brightly stained neurons in the Exoenzyme C3presence group was dramatically decreased as compared with that in the Exoenzyme C3 absence group (P＜0.05), and CCK-8 assay showed that the cell survival rate in Exoenzyme C3 absence group significantly decreased as compared with that in the Exoenzyme C3 presence group (P＜0.05).Conclusion RhoA in membrane is related to the thrombin-induced neuron injury in the cortex of fetal
