生酮饮食调节难治性癫痫患儿外周血辅助性T淋巴细胞亚群平衡的相关性研究

Correction study of ketogenic diet adjustment for balance of peripheral blood helper T cell subsets in children with refractory epilepsy

目的探讨生酮饮食(KD)调节儿童难治性癫痫(CRE)患儿外周血辅助性T淋巴细胞(Th)亚群平衡的相关性。方法回顾性分析2015年1月至2016年5月在郑州大学附属儿童医院接受治疗的CRE患儿42例的临床资料,对所有患儿进行KD治疗,所得检查数据分别纳入CRE患儿治疗前组及CRE患儿治疗后组。另选同期在本院进行健康体检的同龄儿童40例作为健康对照组,比较各组儿童Th17、调节性T淋巴细胞(Treg)及Th1情况、治疗前后的Th17、Treg及Th1细胞有关因子的mRNA表达情况及血浆炎性因子水平。结果CRE患儿的Treg细胞[(1.75±0.53)%]明显少于健康对照组[(3.97±0.28)%],Th1[(12.25±1.03)%]和Th17细胞[(2.89±0.68)%]明显多于健康对照组Th1[(7.75±2.42)%]和Th17细胞[(1.86±0.57)%],差异均有统计学意义(t=23.542、11.049、7.415,均P<0.05)。CRE患儿治疗前组白细胞介素17A(IL-17A)、γ干扰素(IFN-γ)的mRNA表达水平[(2.46±0.75)×10^-4;(1.48±0.64)×10^-2]明显高于对照组[(0.91±0.24)×10^-4;(0.47±0.11)×10^-2];CRE患儿治疗前组细胞毒T淋巴细胞相关抗原4(CTLA-4)及肿瘤坏死因子受体(GITR)的mRNA表达水平[(20.02±6.57)×10^-2;(12.42±6.46)×10^-5]明显低于对照组[(26.57±6.75)×10^-2;(16.31±4.18)×10^-5],差异均有统计学意义(F=4.697、5.232、4.981、3.872,均P<0.05)。CRE患儿治疗后组的IL-17A[(1.20±0.44)×10^-4]、IFN-γ[(0.70±0.41)×10^-2、CTLA-4[(10.72±2.99)×10^-2]及GITR[(6.04±2.51)×10^-5]的mRNA表达水平明显低于CRE患儿治疗前组[(2.46±0.75)×10^-4、(1.48±0.64)×10^-2、(20.02±6.57)×10^-2、(12.42±6.46)×10^-5],差异均有统计学意义(均P<0.05)。CRE患儿治疗前组的IL-17A、IFN-γ、环氧化物酶-2(COX-2)及前列腺素F2α(PGF2α)水平[(26.52±6.17)ng/L、(11.19±3.15)ng/L、(2.14±1.31)ng/L、(205.74±32.30)ng/L]明显高于健康对照组[(13.93±2.98)ng/L、(8.87±1.09)ng/L、(1.04±0.33)ng/L、(109.8±38.74)ng/L],差异均有统计学意义(F=5.361、3.987、3.654、11.370,均P<0.05)。CRE患儿治疗后组的IL-17A[(18.48±6.18)ng/L]、IFN-γ[(9.54±1.42)ng/L]、COX-2[(1.46±0.72)ng/L]及PGF2α[(126.13±13.07)ng/L]水平明显低于CRE患儿治疗前组[(26.52±6.17)ng/L、(11.19±3.15)ng/L、(2.14±1.31)ng/L、(205.74±32.30)ng/L],差异均有统计学意义(均P<0.05)。结论KD对于CRE患儿外周血Th亚群可能有较好的平衡调节作用,且对患儿的Th有关因子及炎性因子水平具有一定的协同变化规律。

Objective To research ketogenic diet(KD) adjustment for the balance of helper T cell subsets in peripheral blood of children with refractory epilepsy(CRE). Methods Forty-two CRE children admitted to Children′s Hospital Affiliated to Zhengzhou University from January 2015 to May 2016 were retrospectively analyzed.All the CRE patients were treated with KD, and the data before and after treatment were collected.During the same period, 40 healthy children were taken as the healthy control group.The changes of the compositions of helper T cells 17(Th17), regulatory T cells (Treg) and helper T cells 1(Th1) in each group were recorded.Meanwhile, mRNA expression of Th17, Treg and Th1 related factors were detected, and plasma levels of inflammatory cytokines were analyzed before and after treatment. Results There were less Treg cells [(1.75±0.53)%] in children with CRE compared with the healthy control group[(3.97±0.28)%], but more Th1[(12.25±1.03)%] and Th17 cells [(2.89±0.68)%] compared with the healthy control group[(7.75±2.42)%, (1.86±0.57)%] (t=23.542, 11.049, 7.415, all P<0.05). The mRNA expression of interleukin-17A (IL-17A), gamma-interferon (IFN-γ), in the CRE group before treatment [(2.46±0.75)×10^-4 (1.48±0.64)×10^-2, were significantly higher than those in the healthy control group [(0.91±0.24)×10^-4 (0.47±0.11)×10^-2. The mRNA expression levels of cytotoxic T lymphocyte associated antigen 4 (CTLA-4) and tumor necrosis factor receptor (GITR) in the pre-treatment group of CRE children[(20.02±6.57)×10^-2 (12.42±6.46)×10^-5 were significantly lower than the healthy control group [(26.57±6.75)×10^-2 (16.31±4.18)×10^-5; the difference was statistically significant (F=4.697, 5.232, 4.981, 3.872, all P<0.05). After treatment, mRNA expression levels of IL-17A [(1.20±0.44)×10^-4, IFN-γ [(0.7±0.41)×10^-2, CTLA-4 [(10.72±2.99)×10^-2 and GITR [(6.04±2.51)×10^-5 were significantly decreased compared with the level of pre-treatment group[(2.46±0.75)×10^-4 (1.48±0.64)×10^-2 (20.02±6.57)×10^-2 (12.42±6.46)×10^-5P<0.05]. The levels of IL-17A, IFN-γ, Cyclooxygenases-2(COX-2) and Prostaglandin F2α (PGF2α) in children with CRE the level of pre-treatment group[(26.52±6.17) ng/L, (11.19±3.15) ng/L, (2.14±1.31) ng/L, (205.74±32.30) ng/L] were significantly higher than those in the healthy control group [(13.93±2.98) ng/L, (8.87±1.09) ng/L, (1.04±0.33) ng/L, (109.80±38.74) ng/L](F=5.361, 3.987, 3.654, 11.370, all P<0.05). The levels of IL-17A[(18.48±6.18) ng/L], IFN-γ [(9.54±1.42 ) ng/L], COX-2 [(1.46±0.72) ng/L] and PGF2α [(126.13±13.07) ng/L]in CRE children were reduced after KD adjustment [(26.52±6.17) ng/L, (11.19±3.15) ng/L, (2.14±1.31) ng/L, (205.74±32.30) ng/L], and the differences were statistically significant (all P<0.05). Conclusions KD adjustment may have a beneficial effect on balance of peripheral blood in children with CRE.KD adjustment is positively correlated with the level of factors related to Th cells and inflammatory cytokines.