基于中医辨证分型的乳腺增生病肠道菌群研究

Research on Intestinal Flora in People with Cyclomastopathy Based on Syndrome Differentiation of Traditional Chinese Medicine

目的探究肝郁痰凝型与冲任失调型乳腺增生病患者肠道菌群结构特征及性激素水平差异,以期为中医证型及性激素与乳腺增生病相关性研究提供肠道微生物层面的依据。方法纳入肝郁痰凝(G)组、冲任失调(C)组及健康对照(N)组各30例,测定血清雌二醇(E_(2))、孕酮(P)及泌乳素(PRL)水平,采集粪便样本,提纯总RNA,用Miseq测序仪进行测序,对测序结果运用R(v3.1.1)软件等进行统计分析。结果(1)G组 E_(2) 水平高于C组及N组(P<0.01),C组 E_(2)水平高于N组(P<0.05);G组及C组P水平均低于N组(P<0.01),PRL水平均高于N组(P<0.01),G组PRL水平高于C组(P<0.01)。(2)G组及C组的observed species、chao及ACE指数均小于N组(P<0.01),G组的shannon指数小于N组(P<0.01),simpson指数大于N组(P<0.05)。(3)各组样品主要构成菌种均为拟杆菌门、厚壁菌门、变形菌门及放线菌门。(4)G组广古菌门、黏胶球形菌门、互养菌门、软壁菌门及疣微菌门相对丰度均低于N组,C组拟杆菌门及梭菌门相对丰度高于N组,互养菌门及疣微菌门相对丰度低于N组,差异有统计学意义(P<0.05)。结论(1) E_(2) 、PRL升高及P降低为乳腺增生病患者特征性性激素水平变化,且肝郁痰凝型患者 E_(2)、PRL升高较冲任失调型显著。(2)肝郁痰凝型及冲任失调型乳腺增生病患者肠道菌群丰度低于健康对照组,且前者肠道菌群多样性亦低于健康对照组。(3)肝郁痰凝型及冲任失调型乳腺增生病患者分别有其特征性肠道菌群。

Objective To explore the structure characteristics of intestinal flora and the difference of sex hormone levels of liver depression and phlegm coagulation type and Chong-Ren disorder type of cyclomastopathy,in order to provide evidence for the correlation among TCM syndrome differentiation,sex hormone and cyclomastopathy at the intestinal flora level.Methods Totally 30 cases with liver depression and phlegm coagulation type,30 cases with chongren disorder type and 30 healthy persons without cyclomastopathy were included separately in group G,group C and group N.The serum levels of estradiol( E_(2)),progesterone(P)and prolactin(PRL)were measured.The fecal samples were collected and the total RNA was purified.The biodiversity sequencing was carried out by Miseq sequencing technology,and the results were analyzed by R(v3.1.1)software.Results(1)The level of E_(2)in group G was higher than that in group C and group N(P<0.01),and that in group C was higher than that in group N(P<0.05),the level of P in group G and group C was lower than that in group N(P<0.01),the level of PRL in group G and group C was higher than that in group N(P<0.01),and the level of PRL in group G was higher than that in group C(P<0.01).(2)The observed species,chao and ACE index of group G and group C were all smaller than those of group N(P<0.01),the shannon index of group G was smaller than that of group N(P<0.01)and the simpson index of group G was larger than that of group N(P<0.05).(3)The main flora were Bacteroidetes,Firmicutes,Proteobacteria and Actinobacteria in each group.(4)The relative abundance of Euryarchaeota,Lentisphaerae,Synergistetes,Tenericutes and Verrucomicrobia in group G were lower than those in group N,the relative abundance of Bacteroidetes and Fusobacteria in group C were higher than those in group N,while the relative abundance of Synergistetes and Verrucomicrobia in group C were lower than that in group N and the difference was statistically significant(P<0.05).Conclusion(1)The increase of E_(2)and PRL and the decrease of P were characteristic changes of sex hormone levels in patients with cyclomastopathy,and the increase of E_(2)and PRL in patients with liver depression and phlegm coagulation type were more significant than that in patients with Chong-Ren disorder type.(2)The intestinal flora richness of patients with liver depression and phlegm coagulation and Chong-Ren disorder type were lower than that of healthy control group and the intestinal flora diversity of the former was lower than that of healthy control group.(3)There were separately characteristic intestinal flora in cyclomastopathy patients with liver depression and phlegm coagulation and Chong-Ren disorder type.