CAJ | 학술논문

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。
목적：탐토타막석분대소효질세포적부함량안산중복서렬격매2（ LRRK2）기인급상관염증인자표체적영향。방법취원대소효질세포분위CON조、지다당（ LPS）조、LPS＋타막석분조、LPS＋LRRK2조；이LPS자격격활소효질세포，용타막석분조공소효질세포격활；장각조소효질세포여대서신상선기락류세포（ PC12）세포공배양24 h；Western Blot법검측소효질세포적LRRK2、일양화담합성매（ iNOS ）표체，매련면역흡부법（ ELISA）측정TNF-α、IL-1β수평；Hochest염핵법검측PC12세포조망/존활비。결과타막석분억제LPS자격후소효질세포적LRRK2표체，통과억제LRRK2하조iNOS、TNF-α급IL-1β석방，진이강저소효질세포여PC12세포공배양체계염증표체급PC12세포조망/존활비。결론 LRRK2시소효질세포염증인자iNOS석방적상유조공파점，타막석분가통과조공LRRK2기인억제소효질세포격활，종이감경염증상관다파알능신경원손상。
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-αand IL-1βby 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 .