中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2015年
2期
142-146
,共5页
周厚福%李成荣%廖建湘%王国兵%林素芳%陈黎
週厚福%李成榮%廖建湘%王國兵%林素芳%陳黎
주후복%리성영%료건상%왕국병%림소방%진려
生酮饮食%过氧化物酶体增殖物激活受体γ%滤泡辅助性T淋巴细胞%白细胞介素-21%B淋巴细胞淋巴瘤6%B淋巴细胞诱导的成熟蛋白-1
生酮飲食%過氧化物酶體增殖物激活受體γ%濾泡輔助性T淋巴細胞%白細胞介素-21%B淋巴細胞淋巴瘤6%B淋巴細胞誘導的成熟蛋白-1
생동음식%과양화물매체증식물격활수체γ%려포보조성T림파세포%백세포개소-21%B림파세포림파류6%B림파세포유도적성숙단백-1
Ketogenic diet%Peroxisome proliferator-activated receptor gamma%Follicular helper T cells%Interleukin-21%B-cell lymphoma 6%B-lymphocyte-induced maturation protein-1
目的 探讨生酮饮食(KD)治疗对难治性癫(痫)患儿滤泡辅助性T淋巴细胞(TFH)的影响.方法 选择2013年7月至2014年1月在深圳市儿童医院住院的难治性癫(痫)患儿33例,男19例,女14例;平均年龄39.6个月.健康对照组选自深圳市儿童医院同期体检健康同龄儿童17例.研究对象分别于KD治疗前和治疗1周时取血备检.流式细胞术检测外周血TFH、各阶段B淋巴细胞比例,酶联免疫吸附试验(ELISA)检测外周血白细胞介素-21(IL-21)水平,实时荧光定量聚合酶链反应(RT-PCR)检测外周血CD4+T淋巴细胞过氧化物酶体增殖物激活受体γ(PPAR-γ)、B淋巴细胞诱导的成熟蛋白-1(Blimp-1)、B淋巴细胞淋巴瘤6(Bcl6)和IL-21 mRNA的表达.结果 1.KD治疗后难治性癫(痫)患儿TFH细胞数量[(3.57±0.58)％]明显下降(P＜0.01),难治性癫(痫)患儿治疗前TFH细胞数量[(4.93±0.70)％]与健康对照组[(5.03±0.63)％]比较差异无统计学意义(P＞0.05).2.难治性癫(痫)患儿KD治疗后TFH细胞转录因子Bcl6表达下降(P＜0.05),抑制因子Blimp-1表达增高(P＜0.05).3.KD治疗后难治性癫(痫)患儿血IL-21水平呈下降趋势,CD4+T淋巴细胞IL-21 mRNA表达较治疗前显著下降(8.28×10-3±1.19×10-3比1.72×10-2±0.81×10-2,t=3.08,P＜0.05).4.KD治疗前后CD27-IgD+B淋巴细胞比较差异无统计学意义(P＞0.05),治疗后CD27+ IgD+B淋巴细胞和CD27-IgD-B淋巴细胞呈下降趋势,但与治疗前比较差异无统计学意义(P＞0.05);CD27+ IgD-B淋巴细胞和CD27+ IgD-CD38 high浆细胞治疗后明显下降(P ＜0.05).5.TFH细胞数量与CD27+ IgD-B淋巴细胞和CD27+IgD-CD38high浆细胞数量呈正相关(r =0.785、0.745,P＜0.05).6.难治性癫(痫)患儿KD治疗后CD4+T淋巴细胞PPAR-γ mRNA表达显著增高(3.49×10-3±1.10×10-3比2.28±10-3±1.30×10-3,t=3.41,P＜0.05),PPAR-γ表达与TFH细胞比例呈负相关(r=-0.619,P＜0.05).结论 KD可通过诱导CD4+T淋巴细胞PPAR-γ表达下调TFH细胞数量及功能,抑制B淋巴细胞分化,这可能是少数患儿长期KD治疗导致低丙种球蛋白血症的原因之一.
目的 探討生酮飲食(KD)治療對難治性癲(癇)患兒濾泡輔助性T淋巴細胞(TFH)的影響.方法 選擇2013年7月至2014年1月在深圳市兒童醫院住院的難治性癲(癇)患兒33例,男19例,女14例;平均年齡39.6箇月.健康對照組選自深圳市兒童醫院同期體檢健康同齡兒童17例.研究對象分彆于KD治療前和治療1週時取血備檢.流式細胞術檢測外週血TFH、各階段B淋巴細胞比例,酶聯免疫吸附試驗(ELISA)檢測外週血白細胞介素-21(IL-21)水平,實時熒光定量聚閤酶鏈反應(RT-PCR)檢測外週血CD4+T淋巴細胞過氧化物酶體增殖物激活受體γ(PPAR-γ)、B淋巴細胞誘導的成熟蛋白-1(Blimp-1)、B淋巴細胞淋巴瘤6(Bcl6)和IL-21 mRNA的錶達.結果 1.KD治療後難治性癲(癇)患兒TFH細胞數量[(3.57±0.58)％]明顯下降(P＜0.01),難治性癲(癇)患兒治療前TFH細胞數量[(4.93±0.70)％]與健康對照組[(5.03±0.63)％]比較差異無統計學意義(P＞0.05).2.難治性癲(癇)患兒KD治療後TFH細胞轉錄因子Bcl6錶達下降(P＜0.05),抑製因子Blimp-1錶達增高(P＜0.05).3.KD治療後難治性癲(癇)患兒血IL-21水平呈下降趨勢,CD4+T淋巴細胞IL-21 mRNA錶達較治療前顯著下降(8.28×10-3±1.19×10-3比1.72×10-2±0.81×10-2,t=3.08,P＜0.05).4.KD治療前後CD27-IgD+B淋巴細胞比較差異無統計學意義(P＞0.05),治療後CD27+ IgD+B淋巴細胞和CD27-IgD-B淋巴細胞呈下降趨勢,但與治療前比較差異無統計學意義(P＞0.05);CD27+ IgD-B淋巴細胞和CD27+ IgD-CD38 high漿細胞治療後明顯下降(P ＜0.05).5.TFH細胞數量與CD27+ IgD-B淋巴細胞和CD27+IgD-CD38high漿細胞數量呈正相關(r =0.785、0.745,P＜0.05).6.難治性癲(癇)患兒KD治療後CD4+T淋巴細胞PPAR-γ mRNA錶達顯著增高(3.49×10-3±1.10×10-3比2.28±10-3±1.30×10-3,t=3.41,P＜0.05),PPAR-γ錶達與TFH細胞比例呈負相關(r=-0.619,P＜0.05).結論 KD可通過誘導CD4+T淋巴細胞PPAR-γ錶達下調TFH細胞數量及功能,抑製B淋巴細胞分化,這可能是少數患兒長期KD治療導緻低丙種毬蛋白血癥的原因之一.
목적 탐토생동음식(KD)치료대난치성전(간)환인려포보조성T림파세포(TFH)적영향.방법 선택2013년7월지2014년1월재심수시인동의원주원적난치성전(간)환인33례,남19례,녀14례;평균년령39.6개월.건강대조조선자심수시인동의원동기체검건강동령인동17례.연구대상분별우KD치료전화치료1주시취혈비검.류식세포술검측외주혈TFH、각계단B림파세포비례,매련면역흡부시험(ELISA)검측외주혈백세포개소-21(IL-21)수평,실시형광정량취합매련반응(RT-PCR)검측외주혈CD4+T림파세포과양화물매체증식물격활수체γ(PPAR-γ)、B림파세포유도적성숙단백-1(Blimp-1)、B림파세포림파류6(Bcl6)화IL-21 mRNA적표체.결과 1.KD치료후난치성전(간)환인TFH세포수량[(3.57±0.58)％]명현하강(P＜0.01),난치성전(간)환인치료전TFH세포수량[(4.93±0.70)％]여건강대조조[(5.03±0.63)％]비교차이무통계학의의(P＞0.05).2.난치성전(간)환인KD치료후TFH세포전록인자Bcl6표체하강(P＜0.05),억제인자Blimp-1표체증고(P＜0.05).3.KD치료후난치성전(간)환인혈IL-21수평정하강추세,CD4+T림파세포IL-21 mRNA표체교치료전현저하강(8.28×10-3±1.19×10-3비1.72×10-2±0.81×10-2,t=3.08,P＜0.05).4.KD치료전후CD27-IgD+B림파세포비교차이무통계학의의(P＞0.05),치료후CD27+ IgD+B림파세포화CD27-IgD-B림파세포정하강추세,단여치료전비교차이무통계학의의(P＞0.05);CD27+ IgD-B림파세포화CD27+ IgD-CD38 high장세포치료후명현하강(P ＜0.05).5.TFH세포수량여CD27+ IgD-B림파세포화CD27+IgD-CD38high장세포수량정정상관(r =0.785、0.745,P＜0.05).6.난치성전(간)환인KD치료후CD4+T림파세포PPAR-γ mRNA표체현저증고(3.49×10-3±1.10×10-3비2.28±10-3±1.30×10-3,t=3.41,P＜0.05),PPAR-γ표체여TFH세포비례정부상관(r=-0.619,P＜0.05).결론 KD가통과유도CD4+T림파세포PPAR-γ표체하조TFH세포수량급공능,억제B림파세포분화,저가능시소수환인장기KD치료도치저병충구단백혈증적원인지일.
Objective To explore the impact of ketogenic diet (KD) on follicular helper T cells(TFH) in children with intractable epilepsy.Methods Thirty-three cases with intractable epilepsy were selected between Jul.2013 and Jan.2014 in Shenzhen Children's Hospital,19 boys and 14 girls; average age was 39.6 months,and seventeen age-matched healthy children who took a physical examination in the same hospital were assigned as the healthy control group.Blood samples were collected from the children with refractory epilepsy before and after 1 week of KD treatment.The proportions of the various stages of B cells and TFH cells were detected by flow cytometry.The plasma concentration of interleukin-21 (IL-21) was determined by enzyme-linked immunosorbent assay(ELISA),and realtime quantitative PCR(RT-PCR) was performed to detect the levels of peroxisome proliferator-activated receptor gamma (PPAR-γ),B-lymphocyte-induced maturation protein-1 (Blimp-1),B-cell lymphoma 6 (Bcl6) and IL-21 mRNA expression in CD4 + T cells.Results (1) The number of TFH cells in children with intractable epilepsy [(3.57 ± 0.58) ％] was remarkably decreased after KD treatment(P ＜ 0.01),while there were no difference between after KD treatment and healthy control group[(4.93 ±0.70)％ vs (5.03 ±0.63)％,P ＞0.05].(2) The levels of transcription factor Bcl6 expression after treatment were significantly decreased,while inhibitory factor Blimp-1 expression increased (P ＜ 0.05).(3)The plasma concentration of IL-21 had a trend to decrease (P ＞ 0.05),while there were no difference before and after KD treatment,and levels of IL-21 mRNA expressions in CD4 +T cells were significantly decreased after the treatment (8.28 × 10-3 ± 1.19 × 10-3 vs 1.72 × 10-2 ± 0.81 × 10-2,t =3.08,P ＜ 0.05).(4) There was no significant difference in CD27-IgD + B cells before and after KD treatment (P ＞ 0.05),CD27 + IgD + B cell and CD27-IgD-B cells had a trend to decrease after KD treatment(P ＞0.05),and CD27 + IgD-B cells and CD27 + IgD-CD38 high plasma cells were significantly decreased after KD treatment (P ＜ 0.05).(5) The number of TFH cells were correlated positively with the number of CD27 + IgD-B cells and CD27 + IgD-CD3g high plasma cells (r =0.785,0.745,P ＜ 0.05).(6) The levels of PPAR-γmRNA in CD4 + T cells expression were significantly up-regulated after KD treatment (3.49 × 10-3 ± 1.10 × 10-3 vs 2.28 ± 10-3 ± 1.30 × 10-3,t =3.41,P ＜0.05),and the number of TFH cells and PPAR-γgene expression was correlated negatively (r =-0.619,P ＜ 0.05).Conclusions KD might down regulate TFH cell number and function through inducing PPAR-γexpression and could inhibit B cell differentiation,which might be one of the factors for hypogammaglobuinemia by KD treatment.
