TGF-β1诱导的Postn、α-SMA和CollagenⅠ在宫腔粘连内膜组织中高表达与粘连严重程度相关

Upregulation of Postn,α-SMA and CollagenⅠ Induced by TGF-β1 was Associated with the Severity of Intrauterine Adhesions

骨膜蛋白(periostin,Postn)与转化生长因子-β1(transforming growth factor-beta1,TGF-β1)信号通路活动密切相关,并参与多种纤维化疾病的病理形成,但在宫腔粘连(intrauterine adhesions,IUAs)形成过程中的作用机制尚不清楚。为了解Postn与纤维标志物(α-平滑肌肌动蛋白:alpha smooth muscle actin,α-SMA,Ⅰ型胶原:CollagenⅠ)在宫腔粘连发病中的作用,本研究收集宫腔粘连(实验组)轻、中、重度各20名患者的内膜组织,非宫腔粘连者20名(对照组),利用实时荧光定量PCR技术、Western印迹技术和免疫组化染色证明:Postn与α-SMA、CollagenⅠ在mRNA和蛋白质表达水平的趋势一致,且与宫腔粘连严重程度呈正相关;实验组Postn、α-SMA和CollagenⅠ较对照组明显升高,其中以中、重度宫腔粘连患者内膜组织Postn、α-SMA和CollagenⅠ表达有非常显著的统计学差异(P<0.01)。同时,为探究TGF-β1与Postn等纤维标志物的关系,利用重组人转化生长因子TGF-β1刺激原代子宫内膜基质细胞后,Western印迹结果显示,TGF-β1与Postn、α-SMA和CollagenⅠ蛋白质表达变化呈现剂量和时间依赖关系。上述结果证实,内膜纤维化是宫腔粘连的主要特征,由Postn等纤维产物参与形成,且与粘连程度相关。推测该病理过程可能与内膜损伤后,TGF-β1促使子宫内膜基质细胞成纤维分化、诱导Postn持续高表达有关。

Periostin（ Postn） has a key role in the transforming growth factor-beta 1（ TGF-β1） signaling pathway and the pathological process of various fibrotic diseases. However,the function of Postn in intrauterine adhesions（ IUAs） was unclear. To investigate the roles of Postn and fibrotic markers（ alpha smooth muscle actin,α-SMA and type Ⅰ Collagen） in IUAs,a total of sixty human endometrial biopsy samples（ mild IUAs： 20,moderate IUAs： 20,severe IUAs： 20） were collected with confirmed IUAs by hysteroscope（ experimental groups）,plus 20 patients without IUAs（ control group）. Quantitative real-time PCR（ qRT-PCR）,Western blotting and immunohistochemistry（ IHC） staining analysis showed that the mRNA and protein expression of Postn was positively correlated with the expression of α-SMA and Collagen Ⅰ and the severity of intrauterine adhesions. We also found that the levels of Postn,α-SMA and Collagen Ⅰin the experimental groups,especially the moderate and severe IUAs groups,were higher than the control group（ P 0. 01）. In order to investigate the relationship between TGF-β1 and fibrosis such as postn,we detected the protein levels of Postn,α-SMA and Collagen Ⅰ in primary endometrial stromal cells（ ESCs）treated with TGF-β1. Western blot analysis revealed that there was a time-and dose-dependent manner between TGF-β1 stimulation and the protein levels of Postn,α-SMA and CollagenⅠ. In contrast,the results confirmed that endometrial fibrosis was the main pathological feature of IUAs,and the up-regulation of Postn participated in the fibrotic process and was related to degree of adhesions. We speculated that the pathomechanism of IUAs in response to endometrial injury may be related to myofibrosis of ESCs,which could be promoted by continuous high expression of Postn upon TGF-β1 stimulation.