单羧酸转运蛋白1在低糖条件下促进M1型小胶质细胞中诱导型一氧化氮合酶表达  

Monocarboxylate transporter 1 enhancing the expression of inducible nitric oxide synthase within M1 phenotype microglia under low-glucose condition

作　　者：于哲成 张晓艳[2] 杜娟娟 周鹏[1] YU Zhe-cheng;ZHANG Xiao-yan;DU Juan-juan;ZHOU Peng(Institute of Neuroscience,Basic Medical College of Wenzhou Medical University,Zhejiang Wenzhou 325035,China;Department of Chinese Traditional Medicine,the Second Affiliated Hospital of Wenzhou Medical University,Zhejiang Wenzhou 325035,China)

机构地区：[1]温州医科大学神经研究所,浙江温州325035 [2]温州医科大学附属第二医院中医系,浙江温州325035

出　　处：《解剖学报》2022年第3期288-294,共7页Acta Anatomica Sinica

基　　金：浙江省自然科学基金(LY19H090009)。

摘　　要：目的探讨在缺血环境下诱导型一氧化氮合酶(iNOS)和单羧酸转运蛋白1(MCT1)与M1型促炎性小胶质细胞的关系。方法小鼠6只,采用光化学栓塞法制作脑缺血模型;BV2细胞使用含5 mmol/L低糖培养基刺激2 h。使用免疫荧光染色、Western blotting方法检测缺血小鼠大脑中小胶质细胞和低糖刺激的BV2细胞中iNOS、MCT1及精氨酸酶1(ARG1)的表达。用RNA干扰及质粒转染技术分别调控未处理BV2细胞中的MCT1表达,检测iNOS及ARG1的变化。结果激光制作脑缺血模型成功后,缺血侧半球中iNOS和MCT1以及ARG1的表达升高(P<0.01),且MCT1主要表达在离子钙接头蛋白分子1(Iba1)阳性和iNOS阳性的小胶质细胞中。低糖刺激BV2小胶质细胞后,iNOS和MCT1表达升高(P<0.001),而ARG1表达降低(P<0.01),MCT1主要表达在iNOS阳性的BV2细胞中。RNA干扰BV2细胞后,细胞中MCT1表达降低,iNOS表达下降(P<0.01);质粒转染BV2细胞后,MCT1表达增高,iNOS表达增加(P<0.01)。但在两种情况下,ARG1表达水平均未发生明显变化。结论在低糖环境中小胶质细胞向M1表型极化,MCT1和iNOS表达同时增高,MCT1参与iNOS的表达上调,可能处于iNOS调节通路的上游。Objective To testify the link between the increase of inducible nitric oxide synthase(iNOS)and monocarboxylate transporter 1(MCT1)expression in the M1 phenotype microglia under ischemic condition.Methods Photothrombosic ischemia stroke model was applied in 6 mice.BV2 cells were treated with low glucose medium contained 5 mmol/L glucose for 2 hours.Immunofluorescent staining and Western blotting were used to check the responses from microglia in the mouse brains subjected to ischemia and BV2 cells were treated with low-glucose treatment.Application of RNA interference or plasmid transfection were used to regulate the MCT1 expression in BV2 cells.Results The expression levels of iNOS,MCT1 and arginase-1(ARG1)increased in the ischemic side compared to the non-ischemic side of mice brain(P<0.01).In the BV2 cells exposed to low-glucose condition,iNOS and MCT1 levels increased(P<0.001),whereas ARG1 level decreased(P<0.01).RNA interference interfered the expression of MCT1 and then decreased the iNOS expression(P<0.01),while overexpression of MCT1 through plasmid transfection increased iNOS expression(P<0.01),while the ARG1 expressions in both conditions were not changed significantly.Conclusion After microglia polarized into inflammatory phenotype during ischemia period,iNOS production is related with MCT1 expression,and MCT1 is in the upstream of iNOS pathway.

关 键 词：诱导型一氧化氮合酶 单羧酸转运蛋白1 缺血性脑卒中 小胶质细胞 光化学栓塞法 免疫印迹法 小鼠 

分 类 号：Q189[生物学—神经生物学]