lncRNA LINC01089对氧糖剥夺/复氧损伤后小胶质细胞极化的影响  

Effect of lncRNA LINC01089 on microglia polarization after oxygen-glucose deprivation/reoxygenation injury

作　　者：陈思思[1] 朱馨艺 邓钢[1] 王龙[1] CHEN Si-si;ZHU Xin-yi;DENG Gang;WANG Long(Department of Neurosurgery,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院神经外科,武汉430060

出　　处：《中国临床神经外科杂志》2022年第3期186-192,共7页Chinese Journal of Clinical Neurosurgery

摘　　要：目的 探讨长链非编码RNA(lncRNA)LINC01089对小胶质细胞氧糖剥夺/复氧(OGD/R)损伤后表型和炎症反应的影响及其机制。方法 体外培养小鼠小胶质细胞B-V2细胞,OGD/R损伤后,转染lncRNA LINC01089过表达质粒及其空载质粒、沉默质粒siRNA及阴性对照,以及miR-449c-5p模拟物、抑制剂及其阴性对照寡核苷酸。采用qRT-PCR检测M1型小胶质细胞标记物iNOS mRNA和CD86 mRNA、M2型小胶质细胞标记物Arg1 mRNA和CD206 MRNA、lncRNA LINC01089、miR-449c-5p的表达水平;采用ELISA检测细胞上清液炎性因子的含量;采用免疫印迹法检测细胞STAT6蛋白表达水平。荧光素酶报告基因实验验证lncRNA LINC01089与miR-449c-5p以及miR-449c-5p与STAT6的靶向关系。结果 OGD/R损伤后,B-V2细胞lncRNA LINC01089表达水平显著下调,miR-449c-5p表达水平显著上调。荧光素酶报告基因实验结果显示,lncRNA LINC01089靶向负调控miR-449c-5p表达,miR-449c-5p靶向负调控STAT6表达。过表达lncRNA LINC01089,显著降低M1型标记物iNOS mRNA和CD86 mRNA表达水平及细胞上清液促炎因子IL-1β、IL-6和TNF-α的含量,明显增高M2型标记物Arg-1 mRNA和CD206mRNA表达水平及细胞上清液抗炎因子IL-4、IL-10和TGF-β的含量。过表达lncRNA LINC01089可靶向负调控B-V2细胞miR-449c-5p表达,促进STAT6蛋白表达。结论 小胶质细胞OGD/R损伤后,lncRNA LINC01089表达下调,促使小胶质细胞向M1型转化,诱导炎症反应,其机制可能与靶向负调控miR-449c-5p/STAT6信号轴有关。Objective To investigate the effect of lncRNA-LINC01089 on the phenotypic polarization of mouse microglia after oxygen-glucose deprivation/reoxygenation(OGD/R) injury. Methods The mouse microglia B-V2 cells were cultured in vitro. After OGD/R injury, lncRNA LINC01089 overexpression plasmid and its empty vector, silencing plasmid siRNA and negative control, as well as miR-449c-5p mimics, inhibitors and its negative control oligonucleotides were transfected into the cells. The expression levels of M1-type microglia markers iNOS and CD86 mRNAs, M2-type microglia markers Arg1 and CD206 mRNAs, lncRNA LINC01089, and miR-449c-5p were detected by qRT-PCR. The contents of inflammatory factors in cell supernatant were detected by ELISA. Western blotting was used to detect the expression level of STAT6 protein in B-V2 cells. The luciferase reporter gene experiment was used to verify the targeting relationship between lncRNA LINC01089 and miR-449c-5p, and miR-449c-5p and STAT6. Results After OGD/R injury, the expression level of lncRNA LINC01089 in B-V2 cells was significantly down-regulated, and the expression level of miR-449c-5p was significantly up-regulated. The results of the luciferase reporter gene assay showed that lncRNA LINC01089 targeted and negatively regulated the expression of miR-449c-5p, and miR-449c-5p targeted and negatively regulated the expression of STAT6.Overexpression of lncRNA LINC01089 significantly decreased the expression levels of M1-type markers iNOS and CD86 mRNAs, and significantly decreased the contents of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in the cell supernatant, and significantly increased the M2-type marker Arg-1 and CD206 mRNAs expression levels, and significantly increased the contents of antiinflammatory factors IL-4, IL-10 and TGF-β in cell supernatants. Overexpression of lncRNA LINC01089 can target and negatively regulate the expression of miR-449c-5p in B-V2 cells and promote the expression of STAT6 protein. Conclusions After OGD/R injury,the expression of lncRNA LI

关 键 词：小胶质细胞 长链非编码RNA lncRNA LINC01089 miR-449c-5p 小胶质细胞极化 

分 类 号：R743[医药卫生—神经病学与精神病学]