肾虚型慢性再障患者肠道菌群结构变化及补肾生血方联合治疗的干预作用

Changes of Intestinal Flora Structure in Patients with Chronic Aplastic Anemia of Kidney Deficiency Type and Intervention Effect of Bushen Shengxue Formula Combined Therapy

摘要: 目的：分析肾虚型慢性再生障碍性贫血(AA)患者的肠道菌群特征，并评估补肾生血方联合免疫抑制治疗对肠道菌群结构的调节作用。方法：本研究将2024年1月至10月于山东中医药大学附属医院确诊的19例肾虚型慢性再障(AA)患者纳入研究。采用自身前后对照设计，在患者接受补肾生血方、环孢素及司坦唑醇的6个月联合治疗前，将其纳入A组(治疗前组)；治疗后，则作为B组(治疗后组)进行数据分析。并招募15例健康受试者作为C组(对照组)。采用Illumina NovaSeq 6000平台对粪便样本的16S rRNA V3~V4区进行测序，并通过QIIME2和R语言进行生物信息学分析，包括Alpha与Beta多样性及LEfSe差异物种筛选。结果：A组(治疗前组)与B组(治疗后组)的肠道菌群多样性存在显著差异(P < 0.05)，而A组与C组(健康对照组)无统计学差异(P > 0.05)。门属水平上，拟杆菌门、变形菌门、厚壁菌门、放线菌门、疣微菌门、埃希氏菌–志贺氏菌属、双歧杆菌属、链球菌属、粪杆菌属和罕见小球菌属为AA的优势菌群。AA患者Patescibacteria菌门、拟杆菌门、乳杆菌属、瘤胃球菌属相对丰度出现上升趋势，弯曲菌门、丹毒梭菌属相对丰度出现下降趋势，与C组间丰度差异具有统计学意义。经补肾生血方联合治疗后，拟杆菌门、Patescibacteria菌门、瘤胃球菌属丰度下降。结论：与健康对照相比，肾虚型慢性再障患者肠道菌群结构发生明显变化，补肾生血方联合治疗可调整其肠道菌群结构达到治疗目的。

Abstract: Objective: To analyze the characteristics of intestinal flora in patients with kidney deficiency-type chronic aplastic anemia (AA) and to evaluate the regulatory effect of Bushen Shengxue Formula combined with immunosuppressive therapy on the intestinal flora structure. Methods: This study enrolled 19 patients with kidney deficiency-type chronic AA diagnosed at the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from January to October 2024. A self-controlled before-and-after design was adopted: before receiving 6 months of combined treatment with Bushen Shengxue Formula, cyclosporine, and stanozolol, the patients were included in Group A (pre-treatment group, n = 19); after treatment, they were analyzed as Group B (post-treatment group, n = 19). Additionally, 15 healthy subjects were recruited as Group C (control group, n = 15). The V3~V4 region of the 16S rRNA gene in fecal samples was sequenced using the Illumina NovaSeq 6000 platform. Bioinformatics analysis was performed using QIIME2 and R software, including alpha diversity, beta diversity, and LEfSe differential species screening. Results: There was a significant difference in intestinal flora diversity between Group A (pre-treatment group) and Group B (post-treatment group) (P < 0.05), whereas no statistically significant difference was observed between Group A and Group C (healthy control group) (P > 0.05). At the phylum and genus levels, Bacteroidetes, Proteobacteria, Firmicutes, Actinobacteria, Verrucomicrobia, Escherichia-Shigella, Bifidobacterium, Streptococcus, Faecalibacterium, and Rarimicrobium were the dominant flora in AA patients. The relative abundance of Patescibacteria, Bacteroidetes, Lactobacillus, and Ruminococcus showed an upward trend in AA patients, while that of Campylobacterota and Erysipelatoclostridium showed a downward trend, with statistically significant differences in abundance compared with Group C. After combined treatment with Bushen Shengxue Formula, the abundance of Bacteroidetes, Patescibacteria, and Ruminococcus decreased. Conclusion: Compared with healthy controls, the intestinal flora structure of patients with kidney deficiency-type chronic AA undergoes significant changes, and combined treatment with Bushen Shengxue Formula can modulate the intestinal flora structure to achieve the therapeutic purpose.