摘要
目的通过甘草次酸(GA)干预X线照射后的小胶质细胞,探究GA对小胶质细胞生物学行为的影响及其机制。方法体外培养小胶质细胞BV2细胞系,分别给予细胞不同的干预措施:甘草次酸1μg/mL+射线照射(GA 1μg/mL+IR组)、甘草次酸10μg/mL+射线照射(GA 10μg/mL+IR组)、空白对照+射线照射(Cont+IR组)、溶剂对照组+射线照射(DMSO+IR组)、地塞米松10 nmol/L+射线照射(Dex10nmol/L+IR组)。照射方法:药物预处理3 h后,以6 MV X射线单次剂量4 GY照射BV2细胞,6 h后检测BV2细胞的ROS含量、IL-1β分泌情况及细胞中Caspase-1、HO-1的表达的变化情况。结果CCK8实验结果显示,BV2细胞经不同浓度GA干预后,GA浓度为10μg/mL时细胞存活率最高达73.876%,1μg/mL时次高为67.226%。流式细胞术结果显示,4 Gy照射后,GA组(1μg/mL)的ROS值较DMSO组明显上调(14.567%vs 7.167%,P=0.025),而与Dex(10 nM)组相比(14.567%vs 12.900%,P=0.995)未见明显改变。ELISA检测结果表明,GA+IR组(GA 10μg/mL组)IL-1β的水平为照射后4组中最低值(0.156),但与Cont+IR组(0.156 vs 0.212,P=0.131)、DMSO+IR组(0.156 vs 0.176,P=0.999)、Dex(10 nM)组(0.156 vs 0.184,P=0.961)相比差异无统计学意义。Western blot检测结果显示GA+IR组的Caspase-1相对蛋白水平较Cont+IR组(0.147 vs 0.243,P=0.000)、DMSO+IR组(0.147 vs 0.590,P=0.000)、Dex+IR组(0.147 vs 0.565,P=0.000)均明显下调,而GA+IR组HO-1相对蛋白水平(0.537)较其他照射组呈上调趋势(P=0.000)。结论GA下调小胶质细胞接受X线照射后Caspase-1蛋白的表达,上调HO-1蛋白水平,表明GA能够抑制细胞凋亡,增加脑内小胶质细胞的抗氧化能力。GA可能在放射性脑损伤过程具有一定的神经保护作用,为临床治疗放射性脑损伤提供可能的治疗机制。
Objective To investigate the effect of glycyrrhetinic acid(GA)on the biological behavior of microglia after X-ray irradiation and its mechanism.Methods Microglia BV2 cell was cultured in vitro and treat⁃ed with different intervention groups:Glycyrrhetinic acid 1μg/mL+irradiation group(GA 1μg/mL+IR group),Glycyrrhetinic acid 10μg/mL+irradiation group(GA 10μg/mL+IR group),Blank control+radiation group(cont+IR group),Solvent control group+radiation group(DMSO+IR group),Dexamethasone 10 nmol/L+radiation group(Dex10nmol/L+IR group).Irradiation method:BV2 cells were irradiated with a dose of 4 Gy 6MV X-ray after 3 hours of drug pretreatment,six hours later,we detected the content of ROS,secretion of IL-1β,expression of caspase-1 and HO-1 in BV2 cells.Results After CCK8 results showed that the survival rate of BV2 cells was 73.876%when GA concentration was 10μg/mL,and 67.226%when GA concentration was 1μg/mL.Flow cytome⁃try results showed that ROS content in GA group(1μg/mL)was significantly up-regulated compared with DMSO group(14.567%vs 7.167%,P=0.025)after 4 Gy irradiation,but no significant differences was observed com⁃pared with Dex group(10 nM)(14.567%vs 12.900%,P=0.995).ELISA results showed that the il-1βlevel in GA+IR group(GA 10μg/mL group)was the lowest(0.156)among the four groups.There was no significant differ⁃ence compared with Cont+IR group(0.156 vs 0.212,P=0.131),DMSO+IR group(0.156 vs 0.176,P=0.999)and Dex(10 nM)group(0.156 vs 0.184,P=0.961).Western blot analysis showed that the relative protein level of Caspase-1 in GA+IR group was higher than that in Cont+IR group(0.147 vs 0.243,P=0.000),DMSO+IR group(0.147 vs 0.590,P=0.000),Dex+IR(0.147 vs 0.565,P=0.000),while the relative protein level of HO-1 in GA+IR group(0.537)was up-regulated compared with other irradiation groups(P=0.000).Conclusion GA can reduce the expression of Caspase-1 protein and raise the level of HO-1 protein in microglia after X-ray irradia⁃tion,which indicated that GA can inhibit apoptosis and increase the antioxida
作者
黎珊
蓝瑞隆
潘晓娴
王彩虹
洪金省
LI Shan;LAN Ruilong;PAN Xiaoxian;WANG Caihong;HONG Jinsheng(Department of Radiotherapy,Cancer Center,The First Affiliated Hospital of Fujian Medical University,Fuzhou,Fujian,350005,China;Key Laboratory of Radiation Biology of Fujian Higher Education Institutions,The First Affiliated Hospital,Fujian Medical University,Fuzhou,Fujian,350005,China)
出处
《中国实用神经疾病杂志》
2022年第6期668-673,共6页
Chinese Journal of Practical Nervous Diseases
基金
福建省卫生厅青年科研基金(编号:2018-1-58)
福建医科大学启航基金项目(编号:2018QH1088)。
关键词
小胶质细胞
甘草次酸
X射线
放射性脑损伤
生物学行为
Microglia
Glycyrrhetinic acid
X-ray
Radiation-induced brain injury
Biological behavior
