目的 分析孤独症谱系障碍(ASD)食物不耐受(FI)患者抗氧化酶、炎性因子、肠道屏障功能及肠道菌群的变化,探讨ASD儿童肠道菌群与食物不耐受程度、抗氧化酶、炎性因子和肠道屏障功能的相关性。方法 选取2023年8月—2024年6月就诊于郑州大学第五附属医院儿童康复科且符合纳入标准ASD儿童65例。依据FI检测结果将ASD儿童分为FI组(35例)及非FI组(N-FI,30例)。酶联免疫吸附试验检测血清炎症因子IL-6、IL-1β和IL-10,抗氧化酶SOD和CAT,肠黏膜屏障D乳酸(D-LA)、二胺氧化酶(DAO)和闭合蛋白(Occludin)。采用 16S rRNA 高通量测序检测患者粪便中肠道菌群的分布情况。结果 FI组血清IL-6、IL-1β、D-LA和DAO水平均高于N-FI组(t=2.815、2.622、2.878、2.049,P<0.05),IL-10、CAT、SOD和Occludin水平均低于N-FI组(t=-2.335、-2.812、-2.526、-2.170,P<0.05)。在属水平,FI组Faecalibacterium、Halomona和 Fusicatenibacter的丰度显著高于N-FI组,Bifidobacterium和Bacteroides的丰度显著低于N-FI组。Pearson相关性分析,FI组的ASD患儿Bifidobacterium与蛋黄/蛋清IgG抗体水平呈负相关(r=0.532);[Ruminococcus]_torques_group与牛奶IgG抗体水平呈正相关(r=0.760);FI组[Eubacterium]_eligens_group与IL-10(r=0.662)和SOD (r=0.536)呈正相关,UBA1819与SOD呈正相关(r=0.574),[Ruminococcus]_torques_group与DLA(r=0.692)和IL-1β(r=0.488)呈正相关,Bifidobacterium与Occludin呈负相关(r=-0.488),差异均有统计学意义(P<0.05)。结论 FI组的ASD患儿肠道菌群与N-FI组患儿之间存在差异, 且肠道菌群的丰度与FI抗体水平、炎症、氧化应激和肠屏障功能之间存在相关性,可能通过靶向肠道菌群加强食物不耐受治疗,有效改善ASD患儿病情。
Abstract
Objective To analyze the changes in antioxidant enzymes, inflammatory cytokines, intestinal barrier function, and gut microbiota in children with autism spectrum disorder (ASD) and food intolerance (FI), in order to explore the correlation of intestinal flora with the degree of FI, antioxidant enzymes, inflammatory cytokines, and intestinal barrier function in ASD children. Methods A total of 65 ASD children who met the inclusion criteria and were admitted to the Children's Rehabilitation Department of the Fifth Affiliated Hospital of Zhengzhou University from August 2023 to June 2024 were selected in this study. The ASD children were divided into the FI group (35 cases) and the non-FI group (N-FI, 30 cases) based on FI test results. Enzyme-linked immunosorbent assay was used to detect serum inflammatory cytokines interleukin (IL)-6, IL-1β, and IL-10, antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), and intestinal mucosal barrier markers D-lactic acid (D-LA), diamine oxidase (DAO), and occludin. 16S rRNA high-throughput sequencing was used to detect the distribution of intestinal flora in patient feces. Results The levels of serum IL-6, IL-1β, D-LA, and DAO in the FI group were significantly higher than those in the N-FI group (t=2.815, 2.622, 2.878, 2.049, P<0.05), while the levels of IL-10, CAT, SOD, and occludin were significantly lower than those in the N-FI group (t=-2.335, -2.812, -2.526, -2.170, P<0.05). At the genus level, the abundances of Faecalibacterium, Halomonas, and Fusicatenibacter in the FI group were significantly higher than those in the N-FI group, while the abundances of Bifidobacterium and Bacteroides were significantly lower. Pearson correlation analysis showed that in the FI group, Bifidobacterium was negatively correlated with egg yolk/egg white IgG antibody levels (r=0.532); [Ruminococcus]_torques_group was positively correlated with milk IgG antibody levels (r=0.760); [Eubacterium]_eligens_group was positively correlated with IL-10 (r=0.662) and SOD (r=0.536), UBA1819 was positively correlated with SOD (r=0.574), [Ruminococcus]_torques_group was positively correlated with D-LA (r=0.692) and IL-1β (r=0.488), and Bifidobacterium was negatively correlated with occludin (r=-0.488,P<0.05). Conclusions There are differences in intestinal flora between ASD children with FI and those without FI, and the abundance of intestinal flora is correlated with FI antibody levels, inflammation, oxidative stress, and intestinal barrier function. Targeting intestinal flora may enhance FI treatment and effectively improve the condition of ASD children.
关键词
孤独症谱系障碍 /
食物不耐受 /
炎性因子 /
氧化应激防御 /
肠道菌群
Key words
autism spectrum disorder /
food intolerance /
inflammatory cytokines /
oxidative stress /
gut microbiota
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基金
河南省科技攻关项目(232102311068;242102311189)
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