壶腹周围憩室对胆总管结石患者结石成分及胆道菌群的影响

探究壶腹周围憩室(periampullary diverticulum, PAD)对胆总管结石成分及胆道菌群的影响。

收集我院行内镜下取石治疗患者44例, 其中14例伴有PAD、30例无PAD, 使用红外光谱法和分光光度法分析结石样本中胆固醇、胆红素含量。从入组患者中分别筛选8例伴有PAD的患者为PAD组, 8例无PAD的为对照组, 收集患者胆汁样本, 提取样本中菌群DNA进行扩增并构建细菌基因文库, 经Illumina MiSeq平台高通量测序后进行生物信息学分析。最后对差异菌群与胆红素、胆固醇进行相关性分析。

2组患者对比年龄、胆管直径、结石直径和总胆红素(TBIL)等资料差异均无统计学意义(均P > 0.05)。2组患者结石中胆固醇和胆红素含量[(39.77±13.00)%vs(61.80±26.00)%, Z=-2.973, P=0.003;(27.03±13.00)%vs(12.27±16.00)%, Z=-2.999, P=0.003)差异存在统计学意义。PAD组与对照组Alpha多样性差异无统计学意义(P > 0.05), 而在物种群落组成上存在差异。卟啉单胞菌属、嗜胆菌属等10种菌属在PAD组中的丰度高于对照组, 真杆菌属、乳杆菌属较对照组降低, 差异均有统计学意义(均P < 0.05)。相较于对照组, PAD组牛磺酸和次牛磺酸的代谢、钙信号通路等5条通路差异存在统计学意义(均P < 0.05)。嗜胆菌属和卟啉单胞菌属与胆红素具有相关性(R²=0.701 3, P < 0.000 1;R²=0.286 8, P=0.032 5)。

伴有PAD的患者结石含有更高的胆红素成分。PAD的存在改变胆道菌群多样性、构成及丰度, 并可能通过菌群变化影响结石成分。

The effect of periampullary diverticulum on the stone composition and biliary flora in patients with choledocholithiasis

ObjectiveTo observe the effect of periampullary diverticulum(PAD) on the composition of common bile duct stones and biliary tract flora.MethodsForty-four patients, 14 with PAD and 30 without, who underwent endoscopic retrograde cholangiopancreatography(ERCP) and lithotripsy at our hospital were enrolled, and the cholesterol and bilirubin components of the stone samples were analyzed qualitatively and quantitatively using infrared spectroscopy and spectrophotometry. Among the enrolled patients, 8 with PAD were selected as the PAD group and 8 without PAD were selected as the control group. Bile samples were collected, and bacterial DNA was extracted from the samples for amplification and construction of bacterial gene library, which was sequenced using Illumina MiSeq high throughput for bioinformatics analysis. Finally, the correlation between differential flora and bilirubin and cholesterol was analyzed.ResultsThere were no significant differences between the two groups in age, bile duct diameter, stone diameter, TBIL, AST, ALT and ALP(all P > 0.05). There were significant differences in cholesterol content(39.77%±13.00% vs 61.80%±26.00%, Z=-2.973, P=0.003) and bilirubin content(27.03%±13.00% vs 12.27%±16.00%, Z=-2.999, P=0.003) in the stones between the two groups. Alpha diversity was not statistically different between the two groups(all P > 0.05), while differences existed in species community composition. Compared with the control group, the abundances of Porphyromonas, Bilophila, Rothia, Streptococcus, Clostridium, Bdellovibrio, Micrococcaceae, and Streptococcaceae increased and those of Eubacterium and Lactobacillus decreased in the PAD group, and the differences were statistically significant(all P < 0.05). Five functional pathways(Taurine and hypotaurine metabolism, Calcium signaling pathway, Biosynthesis and biodegradation of secondary metabolites, Cysteine and methionine metabolism, Lipid metabolism) were significantly different between the two groups(all P < 0.05). Bilophila and Porphyromonas were correlated with bilirubin(R2=0.701 3, P < 0.000 1; R2=0.286 8, P=0.032 5).ConclusionPatients with PAD have higher levels of bilirubin in stone components. PAD alters the diversity, composition and abundance of the biliary flora and may influence the composition of stones.