缺氧条件下CK1δ和PER1可调控BV2细胞的线粒体形态

CK1δand PER1 regulate mitochondrial morphology in BV2 cells under hypoxia

目的本研究拟验证CK1δ、PER1对缺血性脑卒中的调控是否通过调控线粒体的功能实现。方法本实验以小鼠小胶质细胞(BV2细胞)为研究对象,用氯化钴(Co Cl2)处理细胞模拟脑卒中时细胞的缺氧状态,通过抑制、敲低和过表达CK1δ来检测PER1及线粒体的功能。结果 200μmol/L Co Cl2处理BV2细胞24 h后CK1δ和Per1的总体表达水平降低。缺氧条件下,抑制和沉默CK1δ会引起细胞核中PER1蛋白的积累,增加Mfn1的表达量,并抑制Drp1的表达。过表达CK1δ造成Mfn1的表达量降低及Drp1的表达量增加。此外,CK1δ抑制剂组和敲低组线粒体的线状占比增加,而CK1δ过表达组的点状占比明显增加。结论缺氧条件下,CK1δ和PER1可以调控BV2细胞的线粒体形态,进而实现其对缺血性脑卒中的调控。

Objective This study intends to verify whether the regulation of CK1δand PER1 on ischemic stroke may be achieved by regulating the function of mitochondria.Methods In this experiment,mouse microglia(BV2 cells)were used as the research objects.The cells were treated with cobalt chloride(CoCl2)to simulate the hypoxic state of the cells during stroke.The functions of Per1 and mitochondria were detected by inhibiting,knock⁃ing down and overexpressing CK1δ.Results The overall expression levels of CK1δand Per1 in BV2 cells treated with 200μmol/L CoCl2 for 24 h were decreased.Under the hypoxic conditions,the inhibited and silenced CK1δincreased PER1 protein levels in the nucleus,elevated Mfn1 expression,and suppressed Drp1 expression,but the overexpressed CK1δsuppressed Mfn1 expression and elevated Drp1 expression.In addition,the proportion of linear mitochondria was elevated upon CK1δinhibition or silencing,while the proportion of punctate mitochondria was significantly increased upon CK1δoverexpression.Conclusion Under hypoxic conditions,CK1δand PER1 can regulate the mitochondrial morphology of BV2 cells so that ischemic stroke is mediated.