摘要
目的探讨RND1对神经细胞凋亡的作用及可能的相关机制。方法利用脂质体转染质粒,建立体外高表达RND1细胞模型,具体分组为flag-RND1组和flag组;以流式细胞术、Q-PCR和免疫印迹等方法检测各组细胞内RND1、p53、cleaved-Caspase3蛋白表达及凋亡水平。结果与对照组比较,flag-RND1组RND1表达水平升高,流式细胞术显示flag-RND1组细胞凋亡水平增高,同时flag-RND1组细胞内凋亡蛋白cleaved-Caspase3的表达水平较flag组升高(P<0.05);伴随凋亡情况改变,p53蛋白表达水平降低。结论小GTP酶蛋白家族成员RND1可能通过p53蛋白参与调节神经细胞凋亡,这可能是1个新的神经元凋亡调控位点。
Objective To explore the mechanism of RND1 which belongs to small GTPase family regulated neurocytes apoptosis. Methods The plasmid fragment to PC12 cells with liposomes was transfected to build over-expression model. The test groups were performed as follows: flag-RND1 group, flag-group. Flow cytometry, Q-PCR and western blot were performed to test the expressive levels of mRNA, protein, and cell apoptosis. Results The mRNA and protein level of RND1 increased in flag-RND1 group,so dose cleaved-Caspase3,flow cytometry also showed the apoptotic rate increased(P<0.05). Meanwhile, the protein expressive level of p53 decreased. Conclusion RND1, a member of the small GTPase protein family might regulate of neuronal apoptosis through p53 protein, which might be a new neuronal apoptosis regulatory site.
作者
董慧敏
刘宝辉
孙前
吕龙琴
毛善平
Dong Huimin;Liu Baohui;Sun Qian(Department of Neurology,Renmin Hospital of Wuhan University,Wuhan 430060)
出处
《卒中与神经疾病》
2019年第6期649-652,663,共5页
Stroke and Nervous Diseases
基金
国家自然科学基金资助项目(编号为81502175,81371390)