他莫昔芬介导小胶质细胞LRRK-2表达对炎症刺激PC12细胞的影响  

Effect of Tamoxifen on Inflammation-Stimulated PC12 Cells Mediated by LRRK-2 Gene Expression of Microglia

作　　者：卢奎[1] 吴文军[2] 周敏[2] 黎捷[3] 钟健强[3] 张成[1] 

机构地区：[1]中山大学附属第一医院神经科,广州市510080 [2]广东省中山市人民医院神经内科,中山市528403 [3]中山大学附属博济医院神经内科,广州市511300

出　　处：《广西医学》2014年第7期861-864,共4页Guangxi Medical Journal

基　　金：中国博士后科学基金(2013M542232);中山市医学科研基金(2013A020422)

摘　　要：目的探讨他莫昔芬对小胶质细胞的富含亮氨酸重复序列激酶2(LRRK2)基因及相关炎症因子表达的影响。方法取原代小胶质细胞分为CON组、脂多糖(LPS)组、LPS+他莫昔芬组、LPS+LRRK2组;以LPS刺激激活小胶质细胞,用他莫昔芬调控小胶质细胞激活;将各组小胶质细胞与大鼠肾上腺嗜铬瘤细胞(PC12)细胞共培养24 h;Western Blot法检测小胶质细胞的LRRK2、一氧化氮合成酶(iNOS)表达,酶联免疫吸附法(ELISA)测定TNF-α、IL-1β水平;Hochest染核法检测PC12细胞凋亡/存活比。结果他莫昔芬抑制LPS刺激后小胶质细胞的LRRK2表达,通过抑制LRRK2下调iNOS、TNF-α及IL-1β释放,进而降低小胶质细胞与PC12细胞共培养体系炎症表达及PC12细胞凋亡/存活比。结论 LRRK2是小胶质细胞炎症因子iNOS释放的上游调控靶点,他莫昔芬可通过调控LRRK2基因抑制小胶质细胞激活,从而减轻炎症相关多巴胺能神经元损伤。Objective To study the effect of tamoxifen on the expressions of leucine-rich repeat kinase 2（ LRRK-2）gene of microglia and related inflammatory factors. Methods Primary microglia was divided into 4 groups including control group,lipopolysaccharide（ LPS） group,LPS plus tamoxifen group and LPS plus LRRK-2 group. LPS-activated microglia was regulated by tamoxifen. PC12 cells of rats were co-cultured with microglia for 24 hours in each group. The expressions of LRRK-2 gene and iNOS were detected by Western Blot. The levels of tumor necrosis factor α（ TNF-α） and interleukin 1β（ IL-1β） were tested by enzyme-linked immunosorbent assay（ ELISA）. The apoptosis/survival ratio of PC12 cells was determined by Hochest staining. Results Tamoxifen down-regulated the LRRK-2 gene expression of microglia by LPS stimulation,inhibited the release of iNOS,TNF-α and IL-1β by suppressing LRRK-2 gene,and decreased the co-cultured inflammatory expression of microglia and PC12 cells as well as apoptosis /survival ratio of PC12 cells. Conclusion LRRK-2 might be an upstream-regulated target for the release of iNOS in microglia. Tamoxifen can inhibit the activation of microglia by regulation of LRRK-2 gene,which might alleviate the inflammation-associated dopamine neuron injury.

关 键 词：帕金森病 他莫昔芬 富含亮氨酸重复序列激酶2 小胶质细胞 肾上腺嗜铬瘤细胞 

分 类 号：R742.5[医药卫生—神经病学与精神病学]