生酮饮食对难治性癫痫患儿 Th 细胞亚群的影响
생동음식대난치성전간환인 Th 세포아군적영향
Effects of ketogenic diet on helper T cells in patients with intractable epilepsy
  • 万方数据
    中华微生物学和免疫学杂志 중화미생물학화면역학잡지
    CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
    2015年 3期 225-231 ,共7页

    崔萍萍%李成荣%李秋%廖建湘%王国兵%林素芳%陈黎

    최평평%리성영%리추%료건상%왕국병%림소방%진려

    生酮饮食%难治性癫痫%Th细胞亚群%过氧化物酶增殖物激活受体γ%氧化应激 生酮飲食%難治性癲癇%Th細胞亞群%過氧化物酶增殖物激活受體γ%氧化應激
    생동음식%난치성전간%Th세포아군%과양화물매증식물격활수체γ%양화응격
    Ketogenic diet%Intractable epilepsy%Th cells%Peroxisome proliferator-activated re-ceptorγ%Oxidative stress
    目的：探讨生酮饮食（ketogenic diet，KD）治疗对难治性癫痫（intractable epilepsy，IP）患儿Th细胞亚群的影响。方法 IP患儿35例，同年龄健康对照组18例。采用流式细胞术分别检测外周血CD3＋CD8－IFN-γ＋（Th1）细胞、CD3＋CD8－IL-17A＋（Th17）细胞及CD4＋CD25＋Foxp3＋（Treg）细胞比例；实时荧光定量PCR（real-time PCR）检测外周血CD4＋CD25－T细胞中T-bet、ROR-γt、IFN-γ、IL-17A、过氧化物酶增殖物激活受体γ（peroxisome proliferator-activated receptorγ，PPAR-γ）mRNA及CD4＋CD25＋T细胞Foxp3、GITR、CTLA-4、PPAR-γmRNA的表达；酶联免疫吸附试验（ ELISA）检测受试者血浆中前列腺素F2a （prostaglandin F2a，PGF2a）、环氧化酶-2（cyclooxygenases-2，COX-2）蛋白浓度；流式微球阵列技术（CBA）检测受试者血浆IL-17A、IFN-γ蛋白表达水平。结果（1）IP患儿外周血Treg细胞比例明显低于同年龄健康对照组（P＜0．05），KD治疗后，Treg细胞数量明显增加（P＜0．05）；IP患儿外周血Th1、Th17细胞比例明显高于同年龄健康对照组（P＜0．05），KD治疗后显著下降（P＜0．05）；IP患儿外周血CD4＋CD25＋Treg细胞转录因子Foxp3及相关因子GITR、CTLA-4基因转录水平明显低于健康对照组（P＜0．05），治疗后明显上调；CD4＋CD25－T细胞中T-bet、ROR-γt、IL-17A、IFN-γ等Th1／Th17细胞相关因子表达量显著高于健康对照组（ P＜0．05），治疗后明显下降；（2） IP患儿CD4＋CD25＋及CD4＋CD25－细胞PPAR-γ基因表达明显低于对照组（P＜0．05），KD治疗后表达量明显上升（P＜0．05），相关性分析发现Treg细胞与PPAR-γ表达呈正相关（r＝0．61，P＜0．05），Th1及Th17细胞与PPAR-γ表达呈负相关[Th1（r＝－0．54，P＜0．05）；Th17（r＝－0．64，P＜0．05）]；（3）IP患儿IL-17A及IFN-γ血浆浓度明显高于健康对照组（P＜0．05），KD治疗后明显下降（P＜0．05），COX-2及PGF2a 血浆浓度明显高于正常对照组（P＜0．05），KD治疗后显著下降（P＜0．05），相关性分析发现COX-2表达与PPAR-γ呈负相关（r＝－0．571，P＜0．05），PGF2a表达与PPAR-γ亦呈负相关（r＝－0．586，P＜0．05）。结论 KD可抑制氧化应激反应，降低COX-2及PGF2a蛋白浓度，上调PPAR-γ表达，调节Th细胞亚群紊乱，减少炎症细胞因子产生，这可能是KD治疗IP的作用机制之一。
    목적：탐토생동음식（ketogenic diet，KD）치료대난치성전간（intractable epilepsy，IP）환인Th세포아군적영향。방법 IP환인35례，동년령건강대조조18례。채용류식세포술분별검측외주혈CD3＋CD8－IFN-γ＋（Th1）세포、CD3＋CD8－IL-17A＋（Th17）세포급CD4＋CD25＋Foxp3＋（Treg）세포비례；실시형광정량PCR（real-time PCR）검측외주혈CD4＋CD25－T세포중T-bet、ROR-γt、IFN-γ、IL-17A、과양화물매증식물격활수체γ（peroxisome proliferator-activated receptorγ，PPAR-γ）mRNA급CD4＋CD25＋T세포Foxp3、GITR、CTLA-4、PPAR-γmRNA적표체；매련면역흡부시험（ ELISA）검측수시자혈장중전렬선소F2a （prostaglandin F2a，PGF2a）、배양화매-2（cyclooxygenases-2，COX-2）단백농도；류식미구진렬기술（CBA）검측수시자혈장IL-17A、IFN-γ단백표체수평。결과（1）IP환인외주혈Treg세포비례명현저우동년령건강대조조（P＜0．05），KD치료후，Treg세포수량명현증가（P＜0．05）；IP환인외주혈Th1、Th17세포비례명현고우동년령건강대조조（P＜0．05），KD치료후현저하강（P＜0．05）；IP환인외주혈CD4＋CD25＋Treg세포전록인자Foxp3급상관인자GITR、CTLA-4기인전록수평명현저우건강대조조（P＜0．05），치료후명현상조；CD4＋CD25－T세포중T-bet、ROR-γt、IL-17A、IFN-γ등Th1／Th17세포상관인자표체량현저고우건강대조조（ P＜0．05），치료후명현하강；（2） IP환인CD4＋CD25＋급CD4＋CD25－세포PPAR-γ기인표체명현저우대조조（P＜0．05），KD치료후표체량명현상승（P＜0．05），상관성분석발현Treg세포여PPAR-γ표체정정상관（r＝0．61，P＜0．05），Th1급Th17세포여PPAR-γ표체정부상관[Th1（r＝－0．54，P＜0．05）；Th17（r＝－0．64，P＜0．05）]；（3）IP환인IL-17A급IFN-γ혈장농도명현고우건강대조조（P＜0．05），KD치료후명현하강（P＜0．05），COX-2급PGF2a 혈장농도명현고우정상대조조（P＜0．05），KD치료후현저하강（P＜0．05），상관성분석발현COX-2표체여PPAR-γ정부상관（r＝－0．571，P＜0．05），PGF2a표체여PPAR-γ역정부상관（r＝－0．586，P＜0．05）。결론 KD가억제양화응격반응，강저COX-2급PGF2a단백농도，상조PPAR-γ표체，조절Th세포아군문란，감소염증세포인자산생，저가능시KD치료IP적작용궤제지일。
    Objective To investigate the effects of ketogenic diet ( KD) treatment on helper T cell subsets in children with intractable epilepsy( IP) . Methods Thirty-five children with IP and eighteen age-matched healthy subjects were enrolled in this study.The percentages of CD3+CD8-IFN-γ+( Th1 ) cells, CD3+CD8-IL-17A+(Th17) cells and CD4+CD25+Foxp3+(Treg) cells were analyzed by flow cytometry.Re-al-time PCR assay was performed to evaluate the expression of T-bet, ROR-γ, IFN-γ, IL-17A and peroxi-some proliferator activated receptorγ( PPAR-γ) at mRNA level in CD4+CD25-T cells and the transcription-al levels of Foxp3, GITR, CTLA-4 and PPAR-γin CD4+CD25+T cells.The concentrations of cyclooxygen-ases-2 (COX-2) and prostaglandin F2a (PGF2α) in plasma samples were measured by enzyme-linked im-munosorbent assay.The expression of IL-17A and IFN-γin plasma samples were detected by using cytomet-ricbeadarray(CBA).Results (1)ThepercentagesofTregcellsinperipheralbloodsamplesfrompa-tients with IP were lower than those in healthy subjects (P<0.05), which were significantly increased with the treatment of KD (P<0.05).The percentages of Th17 and Th1 cells in patients with IP were significantly higher than those in healthy children (P<0.05), but were remarkably decreased with the treatment of KD (P<0.05).Patients with IP showed decreased transcriptional levels of Foxp3, GITR and CTLA-4 in CD4+CD25+T cells as compared with healthy controls, but were up-regulated with the treatment of KD.The ex-pression of transcription factors including T-bet, ROR-γ, IFN-γand IL-17A in CD4+CD25-T cells in pa-tients with IP were higher than those in healthy subjects and were down-regulated after the treatment of KD (P<0.05).(2) The expression of PPAR-γat mRNA level in CD4+CD25-T and CD4+CD25+T cells were decreased in patients with IP as compared with those in heathy controls, but were increased after the treat-ment of KD (P<0.05).Correlation analysis showed that Treg cells had a positive correlation with PPAR-γ(r=0.61, P<0.05).However, the Th1 and Th17 cells were negatively correlated with PPAR-γ[Th1 (r=-0.54, P<0.05), Th17 (r=-0.64, P<0.05) ].(3) The levels of IL-17A and IFN-γin patients with IP were higher than those in healthy controls, but were decreased with KD treatment (P<0.05).The levels of COX-2 and PGF2αin plasma samples from patients with IP were higher than those in healthy controls ( P<0.05).A negative correlation was observed between COX-2 and PPAR-γ(r=-0.571, P<0.05). Moreover, PGF2αand PPAR-γhad a negative correlation as well (r=-0.586, P<0.05).Conclusion The treatment of KD might enhance the expression of PPAR-γthrough inhibiting the products of oxidative stress such as COX-2 and PGF2α, resulting in the rebalance of Th cell subsets and reduced expression of in-flammatory cytokines.
    원문보기 원문다운로드 사이트로이동
저자의 최근 논문