紫癜性肾炎患儿肠道菌群变化及临床意义研究

doi:10.12114/j.issn.1007-9572.2022.0656

摘要/Abstract

摘要： 背景目前国内外关于过敏性紫癜（HSP）患儿肠道菌群变化的研究数量有限，且尚未见关于紫癜性肾炎（HSPN）患儿疾病早期肠道菌群变化的相关报道。目的探讨HSPN患儿肠道菌群的变化及其在疾病发生、发展中的作用。方法于2019年7—9月选取郑州大学第一附属医院儿科收治的37例HSP初治患儿作为试验组，另外同时选取12例健康志愿儿童作为对照组；并对HSPN患儿随访6个月，根据有无肾损伤进一步分为无肾损伤试验亚组13例和肾损伤试验亚组24例。收集HSP患儿与健康儿童的一般资料及粪便标本，应用高通量测序技术对所有研究对象的肠道菌群进行测序及分析，采用Alpha多样性（Shannon指数、Chao1指数、ACE指数）分析探讨样本内的微生物群落的丰度和多样性，通过主坐标分析（PCoA）来探究不同组别间群落结构的差异，利用线性判别分析及影响因子（LEfSe）分析找到组间差异显著的物种。结果 Alpha多样性分析显示，三组研究对象Shannon指数、Chao1指数、ACE指数比较，差异均无统计学意义（P>0.05）。PCoA显示，三组研究对象肠道菌群群落结构均有差异（P<0.05）；Adonis分析结果显示，无肾损伤试验亚组与对照组肠道菌群群落结构比较，差异有统计学意义（F=2.172，P=0.006）；肾损伤试验亚组与对照组肠道菌群群落结构比较，差异有统计学意义（F=2.217，P=0.006）；无肾损伤试验亚组与肾损伤试验亚组肠道菌群群落结构比较，差异有统计学意义（F=1.590，P=0.045）。LEfSe分析显示，与对照组相比，试验组布劳特氏菌属（Blautia）、金黄杆菌属（Chryseobacterium）、Agathobacter和罗斯伯里氏菌属（Roseburia）丰度显著降低（P<0.05），巨单胞菌属（Megamonas）和肠球菌属（Enterococcus）丰度显著增加（P<0.05）；与无肾损伤试验亚组相比，肾损伤试验亚组纺锤状细菌属（Christensenella）和拟杆菌属（Bacteroides）丰度显著降低（P<0.05），乳杆菌属（Lactobacillus）和罗斯氏菌属（Rothia）丰度显著增加（P<0.05）。结论 HSP患儿存在肠道菌群紊乱，HSPN患儿在疾病早期时的肠道菌群已经与HSP无肾损伤患儿出现差异，疾病早期时的肠道菌群紊乱可能与HSPN的发生有密切联系。

关键词: 紫癜, 过敏性紫癜, 肠道菌群, 菌群结构, 菌群多样性, 儿童, 紫癜性肾炎, 高通量测序技术, 前瞻性研究

Abstract:

Background

The number of studies about the status of intestinal flora in children with Henoch-Sch?nlein purpura (HSP) is limited, and there are no reports on changes of intestinal flora in children with Henoch-Sch?nlein purpura nephritis (HSPN) at the early stage of the disease.

Objective

To investigate the changes of intestinal flora in HSPN children and their association with the development of the disease.

Methods

Thirty-seven newly treated children with HSP (test group) were selected from Department of Pediatrics, the First Affiliated Hospital of Zhengzhou University from July to September 2019, and compared to 12 healthy children (control group) in terms of the status of intestinal flora. The test group was further divided into non-renal injury subgroup (13 cases) and renal injury subgroup (24 cases) according to the prevalence of renal injury during a 6-month follow-up. General data and stool specimens were collected from the affected children and healthy children. High-throughput sequencing was used to sequence and analyze the intestinal flora of all subjects. Alpha diversity (Shannon index, Chao1 index, ACE index) analysis was used to explore the richness and diversity of the microbial communities within the samples. Principal coordinate analysis (PCoA) was used to explore the differences in community structure among the groups, linear discriminant analysis effect size (LEfSe) analysis were used to identify species with significant differences.

Results

Alpha diversity analysis results showed that there was no statistically significant difference in Shannon index, Chao1 index and ACE index among the three groups (P>0.05). PCoA showed that the composition of intestinal flora varied across renal injury and non-renal injury subgroups and the control group (P<0.05). Further Adonis analysis revealed that the intestinal flora composition varied significantly between non-renal injury subgroup and the control group (F=2.172, P=0.006), and between renal injury subgroup and the control group (F=2.217, P=0.006), as well as between renal injury and non-renal injury subgroups (F=1.590, P=0.045). LEfSe analysis showed, compared with the control group, the test group had significantly decreased abundance of Blautia, Chryseobacterium, Agathobacter and Roseburia (P<0.05), and significantly increased abundance of Megamonas and Enterococcus (P<0.05). Compared with non-renal injury subgroup, renal injury subgroup had significantly reduced abundance of Christensenella and Bacteroides (P<0.05), and significantly increased abundance of Lactobacillus and Rothia (P<0.05) .

Conclusion

Intestinal flora disorders were found in HSP children, the intestinal flora of children with HSPN at the early stage of the disease were different from those of HSP children without renal injury, suggesting that the intestinal flora disorder at the early stage of the disease may be closely related to the development of HSPN.

Key words: Purpura, Henoch-Sch?nlein purpura, Intestinal flora, Flora structure, Flora diversity, Child, Henoch-Sch?nlein purpura nephritis, High-throughput sequencing technology, Prospective studies

张莉,张建江,窦文杰,等. 紫癜性肾炎患儿肠道菌群变化及临床意义研究[J]. 中国全科医学, 2023, 26(18): 2250-2255. DOI: 10.12114/j.issn.1007-9572.2022.0656.
ZHANG Li,ZHANG Jianjiang,DOU Wenjie, et al. Intestinal Flora in Children with Henoch-Schönlein Purpura Nephritis: Composition and Abundance Analysis and Clinical Significance[J]. Chinese General Practice, 2023, 26(18): 2250-2255. DOI: 10.12114/j.issn.1007-9572.2022.0656.

图/表 6

表1 三组一般资料比较

Table 1 Comparison of general data of children with Henoch-Sch?nlein purpura with and without renal injury，and control group

组别	例数	性别（男/女）	年龄（±s，岁）	BMI 〔M（P₂₅，P₇₅），kg/m²〕
对照组	12	6/6	8.5±4.4	16.4（15.3，18.4）
无肾损伤试验亚组	13	11/2	8.4±2.9	16.5（15.2，18.7）
肾损伤试验亚组	24	15/9	9.9±3.3	18.0（14.7，20.9）
检验统计量值		3.425^a	-0.974^b	0.847^c
P值		0.178	0.385	0.655

图1 三组稀释曲线

Figure 1 Dilution curves of non-renal injury and renal injury subgroups and the control group

表2 三组Alpha多样性指数比较（±s）

Table 2 Comparison of alpha-diversity indices in non-renal injury and renal injury subgroups and the control group

组别	例数	Shannon指数	Chao1指数	ACE指数
对照组	12	5.0±0.7	279.4±44.3	277.1±45.3
无肾损伤试验亚组	13	4.3±1.4	250.2±73.9	254.7±68.6
肾损伤试验亚组	24	4.3±1.1	264.7±62.3	265.3±57.1
F值		1.779	0.695	0.469
P值		0.180	0.504	0.628

图2 三组肠道菌群结构PCoA注：PCoA=主坐标分析

Figure 2 PCoA analysis of intestinal flora composition in non-renal injury and renal injury subgroups and the control group

图3 试验组与对照组肠道菌群的LEfSe分析注：柱状图显示线性判别分析（LDA）值>4的菌群，即有统计学差异的生物标志物；Clostridia=梭状芽孢杆菌纲，Clostridiales=梭菌属目，Lachnospiraceae=毛螺菌科，Blautia=布劳特氏菌属，Chryseobacterium=金黄杆菌属，Roseburia=罗斯伯里氏菌属，Megamonas=巨单胞菌属，Enterococcus_faecalis=粪肠球菌种，Enterococcus=肠球菌属，Enterococcaceae=肠球菌科

Figure 3 LEfSe analysis of intestinal flora in test group and control group

图4 无肾损伤试验亚组与肾损伤试验亚组肠道菌群的LEfSe分析注：柱状图显示LDA值>4的菌群，即有统计学差异的生物标志物；Bacilli=杆菌纲，Lactobacillales=乳杆菌目，Lactobacillaceae=乳杆菌科，Lactobacillus=乳杆菌属，Rothia=罗斯氏菌属，Lactobacillus_salivarius=唾液乳杆菌，Micrococcaceae=微球菌科，Micrococcales=微球菌亚目，Christensenella=纺锤状细菌属，Bacteroidetes=拟杆菌门，Bacteroidia=拟杆菌纲，Bacteroidales=拟杆菌目，Bacteroidaceae=拟杆菌科，Bacteroides=拟杆菌属

Figure 4 LEfSe analysis of intestinal flora in non-renal injury and renal injury subgroups

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