中华精神科杂志
中華精神科雜誌
중화정신과잡지
CHINESE JOURNA OF PSYCHIATRY
2014年
6期
364-370
,共7页
抑郁症%小胶质细胞%经典型活化%替代型活化%选择性5-羟色胺再摄取抑制剂
抑鬱癥%小膠質細胞%經典型活化%替代型活化%選擇性5-羥色胺再攝取抑製劑
억욱증%소효질세포%경전형활화%체대형활화%선택성5-간색알재섭취억제제
Depression%Microglia%Classical activation%Alternative activation%Selective serotonin reuptake inhibitor
目的 探讨SSRIs氟西汀和艾司西酞普兰对于小胶质细胞不同活化途径的影响.方法 新生2 dSD大鼠脑组织经洗涤、分离、消化、过筛网、离心等处理,获得原代小胶质细胞.将BV2细胞系组和原代细胞组进一步分亚组如下:空白对照组、M1型模型组、M2型模型组、氟西汀组和艾司西酞普兰组,干预24h后使用实时定量聚合酶链式反应(real-time quantitative polymerase chain reaction,RT-PCR)、酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)和免疫印迹法测定相关炎症指标表达水平.结果 (1)M1型活化:RT-PCR示氟西汀可显著降低脂多糖联合干扰素-γ(LPS+INF-γ)诱导的BV2细胞白细胞介素1β(interleukin 1β,IL-1β)、IL-6、肿瘤坏死因子a(tumor necrosis factor,TNF-a)和诱导型一氧化氮合成酶(inducible nitric oxide synthase,iNOS)mRNA表达(均P＜0.05).氟西汀和艾司西酞普兰均可显著抑制LPS+ INF-γ诱导的原代小胶质细胞IL-6(均P＜0.01)、TNF-a(P =0.018、0.029)和iNOS(均P=0.005)mRNA表达.免疫印迹法示氟西汀和艾司西酞普兰可显著抑制LPS+ INF-γ诱导的BV2细胞Iba-1 (P ＜0.01、P=0.002)、CD86(均P＜0.01)蛋白表达.氟西汀和艾司西酞普兰均可显著抑制LPS+ INF-γ诱导的原代小胶质细胞CD86表达(均P＜0.01).ELISA示氟西汀可显著抑制LPS+ INF-γ诱导的BV2细胞TNF-a(P=0.003)和IL-1 β(P=0.002)分泌.氟西汀和艾司西酞普兰可显著抑制LPS+ INF-γ诱导的原代小胶质细胞IL-1β分泌(均P＜0.01).原代小胶质细胞各组间TNF-a分泌差异无统计学意义(F=4.627,P=0.053),进一步组间比较示与模型组[(11.6±1.1)g/L]相比,艾司西酞普兰组[(20.2±1.9) g/L]TNF-a表达增高(P=0.012).(2)M2型活化:RT-PCR示氟西汀可显著提高IL-4诱导的原代小胶质细胞IL-10 mRNA表达(P=0.036).免疫印迹法示氟西汀可显著提高IL-4诱导的BV2细胞(P=0.016)和原代小胶质细胞(P＜0.01)CD206表达.ELISA示氟西汀可显著提高IL-4诱导的BV2细胞(P=0.044)和原代小胶质细胞(P＜0.01)IL-10分泌.结论 氟西汀和艾司西酞普兰可通过影响小胶质细胞活化状态而发挥免疫调节作用,这可能是SSRI类抗抑郁剂发挥抗抑郁作用机制之一.
目的 探討SSRIs氟西汀和艾司西酞普蘭對于小膠質細胞不同活化途徑的影響.方法 新生2 dSD大鼠腦組織經洗滌、分離、消化、過篩網、離心等處理,穫得原代小膠質細胞.將BV2細胞繫組和原代細胞組進一步分亞組如下:空白對照組、M1型模型組、M2型模型組、氟西汀組和艾司西酞普蘭組,榦預24h後使用實時定量聚閤酶鏈式反應(real-time quantitative polymerase chain reaction,RT-PCR)、酶聯免疫吸附測定(enzyme linked immunosorbent assay,ELISA)和免疫印跡法測定相關炎癥指標錶達水平.結果 (1)M1型活化:RT-PCR示氟西汀可顯著降低脂多糖聯閤榦擾素-γ(LPS+INF-γ)誘導的BV2細胞白細胞介素1β(interleukin 1β,IL-1β)、IL-6、腫瘤壞死因子a(tumor necrosis factor,TNF-a)和誘導型一氧化氮閤成酶(inducible nitric oxide synthase,iNOS)mRNA錶達(均P＜0.05).氟西汀和艾司西酞普蘭均可顯著抑製LPS+ INF-γ誘導的原代小膠質細胞IL-6(均P＜0.01)、TNF-a(P =0.018、0.029)和iNOS(均P=0.005)mRNA錶達.免疫印跡法示氟西汀和艾司西酞普蘭可顯著抑製LPS+ INF-γ誘導的BV2細胞Iba-1 (P ＜0.01、P=0.002)、CD86(均P＜0.01)蛋白錶達.氟西汀和艾司西酞普蘭均可顯著抑製LPS+ INF-γ誘導的原代小膠質細胞CD86錶達(均P＜0.01).ELISA示氟西汀可顯著抑製LPS+ INF-γ誘導的BV2細胞TNF-a(P=0.003)和IL-1 β(P=0.002)分泌.氟西汀和艾司西酞普蘭可顯著抑製LPS+ INF-γ誘導的原代小膠質細胞IL-1β分泌(均P＜0.01).原代小膠質細胞各組間TNF-a分泌差異無統計學意義(F=4.627,P=0.053),進一步組間比較示與模型組[(11.6±1.1)g/L]相比,艾司西酞普蘭組[(20.2±1.9) g/L]TNF-a錶達增高(P=0.012).(2)M2型活化:RT-PCR示氟西汀可顯著提高IL-4誘導的原代小膠質細胞IL-10 mRNA錶達(P=0.036).免疫印跡法示氟西汀可顯著提高IL-4誘導的BV2細胞(P=0.016)和原代小膠質細胞(P＜0.01)CD206錶達.ELISA示氟西汀可顯著提高IL-4誘導的BV2細胞(P=0.044)和原代小膠質細胞(P＜0.01)IL-10分泌.結論 氟西汀和艾司西酞普蘭可通過影響小膠質細胞活化狀態而髮揮免疫調節作用,這可能是SSRI類抗抑鬱劑髮揮抗抑鬱作用機製之一.
목적 탐토SSRIs불서정화애사서태보란대우소효질세포불동활화도경적영향.방법 신생2 dSD대서뇌조직경세조、분리、소화、과사망、리심등처리,획득원대소효질세포.장BV2세포계조화원대세포조진일보분아조여하:공백대조조、M1형모형조、M2형모형조、불서정조화애사서태보란조,간예24h후사용실시정량취합매련식반응(real-time quantitative polymerase chain reaction,RT-PCR)、매련면역흡부측정(enzyme linked immunosorbent assay,ELISA)화면역인적법측정상관염증지표표체수평.결과 (1)M1형활화:RT-PCR시불서정가현저강저지다당연합간우소-γ(LPS+INF-γ)유도적BV2세포백세포개소1β(interleukin 1β,IL-1β)、IL-6、종류배사인자a(tumor necrosis factor,TNF-a)화유도형일양화담합성매(inducible nitric oxide synthase,iNOS)mRNA표체(균P＜0.05).불서정화애사서태보란균가현저억제LPS+ INF-γ유도적원대소효질세포IL-6(균P＜0.01)、TNF-a(P =0.018、0.029)화iNOS(균P=0.005)mRNA표체.면역인적법시불서정화애사서태보란가현저억제LPS+ INF-γ유도적BV2세포Iba-1 (P ＜0.01、P=0.002)、CD86(균P＜0.01)단백표체.불서정화애사서태보란균가현저억제LPS+ INF-γ유도적원대소효질세포CD86표체(균P＜0.01).ELISA시불서정가현저억제LPS+ INF-γ유도적BV2세포TNF-a(P=0.003)화IL-1 β(P=0.002)분비.불서정화애사서태보란가현저억제LPS+ INF-γ유도적원대소효질세포IL-1β분비(균P＜0.01).원대소효질세포각조간TNF-a분비차이무통계학의의(F=4.627,P=0.053),진일보조간비교시여모형조[(11.6±1.1)g/L]상비,애사서태보란조[(20.2±1.9) g/L]TNF-a표체증고(P=0.012).(2)M2형활화:RT-PCR시불서정가현저제고IL-4유도적원대소효질세포IL-10 mRNA표체(P=0.036).면역인적법시불서정가현저제고IL-4유도적BV2세포(P=0.016)화원대소효질세포(P＜0.01)CD206표체.ELISA시불서정가현저제고IL-4유도적BV2세포(P=0.044)화원대소효질세포(P＜0.01)IL-10분비.결론 불서정화애사서태보란가통과영향소효질세포활화상태이발휘면역조절작용,저가능시SSRI류항억욱제발휘항억욱작용궤제지일.
Objective To explore the effect of the selective serotonin reuptake inhibitors (SSRIs):fluoxetine and escitalopram on the activation of microglia.Methods The experiment involved two groups of cells,the cell line (BV2) and primary microglia cell,and each group was divided into five sub-groups,the control group,M1 model group,M2 model group,the fluoxetine group and the escitalopram group.After intervention for 24 hours the inflammation index were measured by RT-PCR,enzyme linked immunosorbent assay(ELISA) and Western blot.Results (1) M1 activation:Detected by RT-PCR,fluoxetine significantly reduced the LPS + INF-γinduced expressions of IL-1 β (P =0.001),IL-6 (P ＜ 0.01),tumor necrosis factor (TNF-a) (P =0.016) and inducible nitric oxide synthase (iNOS) (P =0.005) mRNA for BV2.Both fluoxetine and escitalopram significantly reduced the LPS + INF-γ induced expressions of IL-6 (P ＜0.01 respectively),TNF-a (P =0.018,0.029 respectively) and iNOS (P =0.005 respectively) mRNA for primary cells.Detected by Western blot,both fluoxetine and escitalopram significantly reduced LPS + INF-γ induced expressions of Iba-1 (P ＜ 0.01,P =0.002 respectively) and CD86 (P ＜ 0.01 respectively) for BV2 cells.For primary microglia cells,both fluoxetine and escitalopram significantly reduced LPS + INF-γinduced expressions of CD86 (P ＜0.01 respectively).Detected by ELISA,fluoxetine significantly reduced the LPS + INF-γ induced releases of TNF-a (P =0.003) and IL-1β (P =0.002) for BV2 cells.For primary microglia cells,both fluoxetine and escitalopram significantly reduced the LPS + INF-γ induced releases of IL-1 β (P ＜ 0.01 respectively).The TNF-a releases of escitalopram group ((20.2 ± 1.9) g/L were significantly higher than the model group (11.6 ± 1.1) g/L,P =0.012).(2)M2 activation:Detected by RT-PCR,fluoxetine significantly improved the IL-4 induced IL-10mRNA expressions for primary cells (P =0.036).Detected by Western blot,fluoxetine significantly improved the IL-4 induced CD206 expressions for BV2 cells (P =0.016) and primary cells (P ＜ 0.01).Detected by ELISA,fluoxetine significantly improved the IL-4 induced IL-10 releases for BV2 cells (P =0.044) and primary cells (P ＜ 0.01).Conclusions Fluoxetine and escitalopram play a role in immune regulation by affecting the activation of microglia,which might be one of the mechanisms of the SSRIs treatment.
