School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
J Tradit Chin Med. 2020 Feb;40(1):137-143.
To investigate the fecal microbiota changes in patients with spleen (Pi)-deficiency (SPD), a common Chinese medicine syndrome with digestive and absorptive disturbances and to provide insight into the relationship between Chinese medicine syndrome and gut dysbiosis.
Fecal microbiotas from the stool samples of 53 SPD patients and 35 healthy subjects were analyzed via 16S rRNA gene polymerase chain reaction (PCR)-denaturation gradient gel electrophoresis (DGGE). SPD-related marker genes from 20 SPD patients and 20 healthy subjects were identified through gene sequencing, while some genes were quantified using quantitative PCR (qPCR). Discriminant analysis was conducted using SPSS software, and the canonical discriminant function formula for Pi-deficiency was established.
Alterations in microbiota diversity and composition between the SPD and healthy groups were demonstrated via 16S rRNA gene PCR-DGGE combined with multivariate statistical analysis. Fecal microbiota changes were also observed among different SPD-subtype patients. Eight SPD-related markers were found, and putative species corresponding to these markers were identified through gene sequencing, which may have potential associations with the common digestive dysfunctions in SPD patients. qPCR methods were established for two of these markers, which were significantly altered in the SPD patients. The canonical discriminant function formula was calculated for SPD, and the validity rates of these markers were over 85%.
Fecal microbiotas are altered in patients with SPD, which may provide insight for further studies on clinically diagnosing and treating SPD. The results may also provide data to gain a better understanding of Traditional Chinese Medicine syndrome and gut dysbiosis.
研究脾虚证（SPD）患者粪便微生物群的变化，脾虚证是一种常见的中医证候，伴有消化和吸收障碍，并深入了解中医证候与肠道菌群失调的关系。
采用 16S rRNA 基因聚合酶链反应（PCR）-变性梯度凝胶电泳（DGGE）分析 53 例 SPD 患者和 35 例健康对照者的粪便微生物群。通过基因测序从 20 例 SPD 患者和 20 例健康对照者中鉴定 SPD 相关标记基因，同时使用定量 PCR（qPCR）定量部分基因。使用 SPSS 软件进行判别分析，并建立脾虚证的典型判别函数公式。
16S rRNA 基因 PCR-DGGE 结合多变量统计分析表明，SPD 组和健康组之间的微生物多样性和组成发生了变化。不同 SPD 亚型患者的粪便微生物群也发生了变化。发现了 8 个 SPD 相关标记物，并通过基因测序鉴定了这些标记物对应的假定种，这些种可能与 SPD 患者常见的消化功能障碍有关。为其中两个标记物建立了 qPCR 方法，这些标记物在 SPD 患者中明显改变。计算了 SPD 的典型判别函数公式，这些标记物的有效率超过 85%。
SPD 患者粪便微生物群发生改变，这可能为进一步研究 SPD 的临床诊断和治疗提供依据。结果还可能为更好地理解中医证候和肠道菌群失调提供数据。
