基于microRNA表达的多囊卵巢综合征患者痰湿证生物标志物的初筛

Screening the biomarkers of phlegm-dampness syndrome in patients with polycystic ovary syndrome based on microRNA expression

目的探索多囊卵巢综合征(PCOS)患者痰湿证的特征性生物标志物,为中医证候诊断的标准化提供理论依据。方法采用队列研究的方法,收集2018年1月至2019年1月期间在山东中医药大学附属医院行体外受精(IVF)助孕治疗,年龄在20~37岁之间的不孕症患者。将患者分为对照组(A组,20例)、PCOS痰湿组(B组,20例)、PCOS非痰湿组(C组,20例)。采用qRT-PCR法检测3组患者血清、卵泡液、颗粒细胞中miR-183/200/223的表达水平,并用相关性分析评价其与痰湿证的关系。结果血清中miR-223-3p在B组的相对表达量高于A组和C组,差异均有统计学意义(P均<0.001),并与胰岛素抵抗指数(HOMA-IR)呈正相关(r=0.595,P=0.04)。卵泡液中miR-223-3p在B组的相对表达量高于A组(P<0.001)及C组(P=0.002),差异均有统计学意义。颗粒细胞中miR-183-5p在B组的相对表达量均高于A组(P=0.004)和C组(P=0.012),差异均有统计学意义,并与HOMA-IR呈正相关(r=0.732,P=0.007);miR-200c-3p在B组的相对表达高于A组(P=0.002)和C组(P=0.001),差异均有统计学意义,并与HOMA-IR呈正相关(r=0.704,P=0.016);miR-223-3p在B组的相对表达量高于A组(P<0.001),在C组的相对表达量也高于A组(P=0.005),差异均有统计学意义,且在B组的相对表达量与HOMA-IR呈正相关(r=0.547,P=0.042);miR-223-5p在B组的相对表达量明显低于A组(P=0.001),在C组的相对表达量也低于A组(P<0.001),差异均有统计学意义,且B组的相对表达量与HOMA-IR呈负相关(r=-0.671,P=0.024)。结论PCOS痰湿证有明显的胰岛素抵抗特征,microRNA在PCOS痰湿证患者体内表达存在显著性差异。miR-183-5p、miR-200c-3p与miR-223-5p可能成为诊断PCOS痰湿证卵巢局部的特异性生物标志物。

Objective To explore the characteristic biomarkers of phlegm-dampness syndrome in patients with polycystic ovary syndrome(PCOS)to provide a theoretical basis for the standardization of Traditional Chinese Medicine(TCM)syndrome diagnosis.Methods This study used the method of cohort study.From January 2018 to January 2019,female infertility patients aged between 20 years and 37 years were collected and treated with in vitro fertilization(IVF)in the Affiliated Hospital of Shandong University of Traditional Chinese Medicine.The cases were divided into control group(group A,n=20),PCOS phlegm and dampness group(group B,n=20),and PCOS non-phlegm and dampness group(group C,n=20).qRT-PCR was used to detect the expression levels of miR-183/200/223 in serum,follicular fluid and granulosa cells among the three groups,and the relationship between miR-183/200/223 and phlegm-dampness syndrome was evaluated by correlation analysis.Results In serum,the relative expression of miR-223-3p in group B was higher than that in group A(P<0.001)and group C(P<0.001),and the differences were statistically significant,and it was positively correlated with homeostasis model assessment of insulin resistance(HOMA-IR)(r=0.595,P=0.04).In follicular fluid,the relative expression of miR-223-3p in group B was higher than that in group A(P<0.001)and group C(P=0.002),and the differences were statistically significant.In granulosa cells,the relative expression of miR-183-5p in group B was higher than that in group A(P=0.004)and group C(P=0.012),and the differences were statistically significant,and it was positively correlated with HOMA-IR(r=0.732,P=0.007);the relative expression of miR-200c-3p in group B was higher than that in group A(P=0.002)and group C(P=0.001),and the differences were statistically significant,and it was positively correlated with HOMA-IR(r=0.704,P=0.016);the relative expression of miR-223-3p in group B was higher than that in group A(P<0.01),and the relative expression in group C was also higher than that in group A(P=0.005),and the relative expression of group B was positively correlated with HOMA-IR(r=0.547,P=0.042);the relative expression of miR-223-5p in group B was significantly lower than that in group A(P=0.001),and the relative expression in group C was also lower than that in group A(P<0.001),and the differences were statistically significant,and the relative expression of group B was negatively correlated with HOMA-IR(r=-0.671,P=0.024).Conclusion PCOS phlegm-dampness syndrome has obvious insulin resistance characteristics,and microRNA expression in PCOS phlegm-dampness syndrome patients has significant differences.miR-183-5p,miR-200c-3p and miR-223-5p may become specific biomarkers for the diagnosis of PCOS phlegm-dampness syndrome.