刺五加对颈性眩晕患者多巴胺D2受体基因CpG岛甲基化及体外小胶质细胞的M1/M2极性的影响

目的 研究刺五加对颈性眩晕(CV)患者多巴胺D2受体(DRD2)基因CpG岛甲基化及体外小胶质细胞hmc-3的M1/M2极性的影响.方法 收集30例未经治疗的CV患者、30例经刺五加治疗的CV患者、30例健康者的血液,样本分别分为CV组、治疗组、健康组.培养hmc-3小胶质细胞,分别应用20μg、40μg、80μg刺五...

Effects of Acanthopanax senticosus on CpG island methylation of dopamine D2 receptor gene and M1/M2 polarity of microglia in vitro in patients with cervical vertigo

Objective To study the effect of Acanthopanax senticosus on the CpG island methylation of dopamine D2 receptor(DRD2)gene and the M1/M2 polarity of microglia hmc-3 in vitro in patients with cervical vertigo(CV).Methods The blood of 30 untreated CV patients,30 CV patients treated with Acanthopanax senticosus,and 30 healthy individuals was collected.The samples were divided into CV group,treatment group,and healthy group.Hmc-3 microglia were cultured and treated with 20μg,40μg,and 80μg Acanthopanax senticosus respectively.The cells were divided into control group and acanthopanax group.The blood whole genome DNA and total RNA in blood or microglia were extracted.The mRNA levels of DRD2,M1 phenotype markers CD16,CD32,CD86 and M2 phenotype marker Arginase 1 and CD206 were detected by RT-qPCR.The DRD2 methylation of CV group,treatment group,healthy group,control group and Acanthopanax senticosus group was detected by sequencing method after modification with bisulfite.CCK-8 method was used to detect the change in the proliferation rate of hmc-3 cells.Western blot was used to detect the protein levels of DRD2,CD16,CD32,CD86,Arginase 1,and CD206.Cellular immunofluorescence method was used to detect the expression of DRD2 in hmc-3 cells.Results Compared with the healthy group,the mRNA expression of DRD2 in CV group was down-regulated,while the methylation rate was up-regulated(P<0.05);compared with CV group,the mRNA expression of treatment group was up-regulated,while the methylation rate was down-regulated(P<0.05).Acanthopanax senticosus 40μg and 80μg up-regulated the proliferation rate and DRD2 expression of microglia hmc-3(P<0.05).Compared with control group,Acanthopanax senticosus down-regulated the mRNA and protein levels of M1 phenotype CD16,CD32 and CD86(P<0.05),while it up-regulated the expression of M2 phenotype Arginase 1 and CD206(P<0.05).Conclusion Acanthopanax senticosus inhibits the methylation rate of CpG island of DRD2 gene in CV patients and promotes the M2 polar phenotype of microglia in vitro.