楂曲消风散治疗儿童特应性皮炎湿热蕴结证的临床疗效及其对皮损处皮肤菌群的影响

Clinical Efficacy and Regulation of Skin Microbiota in Children with Atopic Dermatitis and Damp-heat Accumulation Syndrome Treated with Zhaqu Xiaofeng Powder(楂曲消风散)

目的评价楂曲消风散治疗儿童特应性皮炎(AD)湿热蕴结证的临床疗效及其对皮肤菌群的调控作用。方法采用随机对照试验设计,将90例患儿随机分为治疗组和对照组各45例。治疗组内服楂曲消风散,对照组内服盐酸西替利嗪滴剂,疗程均为4周。分别记录治疗前及治疗2、4周两组患儿特应性皮炎积分指数(SCORAD)评分、瘙痒视觉模拟量表(VAS)评分、儿童皮肤病生活质量指数问卷(CDLQI)评分、中医证候(心烦、口渴、纳呆、小便短赤、大便干或黏)评分。分别于两组中各随机选择25例患儿,在治疗前后进行皮损处皮肤取样,提取皮肤样本DNA进行16S rRNA基因扩增子测序,检测皮肤菌群水平,对菌种分布、菌群多样性及组间差异进行比较。结果治疗组与对照组各脱落5例,最终两组各40例纳入分析。与治疗前比较,两组治疗2、4周SCORAD评分、VAS评分、CDLQI评分均较治疗前降低,且治疗4周改善程度均优于治疗2周(P<0.01)。治疗4周,治疗组SCORAD评分、VAS评分、CDLQI评分显著低于对照组(P<0.01)。与治疗前比较,治疗组心烦、口渴、纳呆、小便短赤、大便干或黏评分均降低(P<0.05);对照组口渴、纳呆、小便短赤评分均降低(P<0.05)。治疗后治疗组心烦、纳呆、小便短赤、大便干或黏评分低于对照组(P<0.05)。两组治疗前后菌种相对丰度比较,对照组缓症链球菌、大肠杆菌、金黄色葡萄球菌、溶血孪生球菌相对丰度下调,对照组缓症链球菌相对丰度下调(P<0.05)。治疗后治疗组金黄色葡萄球菌、缓症链球菌、大肠杆菌、溶血孪生球菌相对丰度低于对照组,且差异有统计学意义(P<0.05)。Alpha多样性分析发现,与治疗前比较,治疗组治疗后Chaol指数、Shannon指数均升高(P<0.05),对照组治疗前后Chaol指数、Shannon指数差异无统计学意义(P>0.05),且治疗后治疗组Chaol指数、Shannon指数均高于对照组(P<0.05)。Beta多样性分析显示,两组患儿治疗前皮损部位菌群结构差异较大,治疗后治疗组菌群结构相似,对照组与治疗前菌群结构无明显变化。菌属结构差异分析显示,两组治疗前后关键菌属均有显著性差异,且两组关键菌属亦不同。结论楂曲消风散可有效减轻AD湿热蕴结证患儿皮损严重程度、缓解皮肤瘙痒情况、提高患儿生活质量,其机制可能通过减少致病菌定植、增加有益菌定植,改变皮肤微生态发挥作用。

Objective To assess the clinical efficacy and regulation of skin microbiota in children with atopic dermatitis and damp-heat accumulation syndrome treated by Zhaqu Xiaofeng Powder(楂曲消风散,ZXP).Methods Ninety children were randomized into a treatment group and a control group,each with 45 children.The treatment group received ZXP orally,while the control group received levocetirizine hydrochloride syrup,both for 4 weeks.The atopic dermatitis severity index(SCORAD)score,visual analog scale(VAS)score for itching,children dermatology life quality index(CDLQI)score,and traditional Chinese medicine syndrome score were assessed before and after 2-and 4-week treatment.Simultaneously,adhering to the principles of sample size in microbial sequencing,25 children were randomly selected from each group(total 50 children);skin samples were collected before and after treatment,and skin specimen DNA was extracted for 16S rRNA gene amplifier sequencing;the skin microbiota levels were detected,and the distribution of bacteria,diversity of flora,and differences between groups were compared.Results There were five drop-outs in each group,and 40 cases in each group were included in final analysis.After 2-and 4-week treat⁃ment,both groups showed a significant reduction in SCORAD scores,VAS scores,and CDLQI scores,and more re⁃ductions were shown after 4-week treatment than 2-week treatment(P<0.01).The SCORAD score,VAS score,and CDLQI score of the treatment group were significantly lower than those in the control group after 4-week treat⁃ment(P<0.01).The scores of upset,thirsty,poor appetite,short red urine,and dry stool were reduced in the treatment group(P<0.05),while the scores of thirsty,poor appetite,short and red urine decreased after treatment(P<0.05).After 4 weeks of treatment,among the differential genera with abundances>0.5%in the treatment group,The cumulative relative abundances of Staphylococcus aureus,Streptococcus_mitis,Escherichia coli and Gemella_haemolysans in the treatment group were downregulated after treatment;in the control group,there was a relative cumulative decrease in the abundance of Streptococcus_mitis.The control group had reduced relative abun⁃dance of Streptococcus_mitis,Escherichia coli,Staphylococcus aureus and Gemella_haemolysans,after treatment(P<0.05).Alpha diversity analysis revealed an increase in both Chaol index and Shannon index after treatment(P<0.05),while there was no significant difference in Chaol index and Shannon index in the control group before and after treatment(P>0.05).Higher Chaol index and Shannon index were found in the treatment group(P<0.05).Beta diversity analysis showed that there were significant differences in the microbial community structure at the lesion site between the treatment group and the control group before treatment.The microbial community structure in the treatment group was similar after treatment,while there was no significant change in the microbial community struc⁃ture in the control group before and after treatment.There were significant structure differences of key bacteria genus in both groups before and after treatment.Conclusion ZXP used in the treatment of pediatric atopic dermatitis(AD)with the syndrome of damp-heat accumulation,has shown efficacy in reducing the severity of skin lesions,alle⁃viating itching,and enhancing the quality of life in children.This modulation aims to decrease the abundance of pathogenic bacteria while promoting the colonization of beneficial bacteria,thereby altering the skin microbiota and contributing to the treatment of pediatric AD.