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腺病毒介导的环氧化酶-2反义表达逆转人黏附性成纤维细胞的异常基因谱。

Adenovirus-mediated expression of cyclooxygenase-2 antisense reverse abnormal genetic profile of human adhesion fibroblasts.

作者信息

Saed Ghassan M, Al-Hendy Ayman, Salama Salama A, Diamond Michael P

机构信息

Department of Obstetrics and Gynecology, C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.

出版信息

Fertil Steril. 2008 May;89(5 Suppl):1455-60. doi: 10.1016/j.fertnstert.2007.04.061. Epub 2007 Jul 10.

Abstract

OBJECTIVE

To determine the effects of blocking the translation of cyclooxygenase-2 (COX-2) mRNA on the mRNA levels of type I collagen, type III collagen, fibronectin, and transforming growth factor-beta (TGF-beta1) in fibroblasts obtained from normal peritoneal and adhesion tissues.

DESIGN

Prospective experimental study.

SETTING

University medical center.

PATIENT(S): Fibroblasts established from peritoneal and adhesion tissue of the same patients.

INTERVENTION(S): Adenovirus with COX-2 treatment of the primary cultured fibroblasts.

MAIN OUTCOME MEASURE(S): Fibroblasts of normal peritoneal and adhesion tissues were isolated from the same patients. Adhesion and normal peritoneal fibroblasts were transfected with an adenovirus COX-2 mRNA in sense or antisense orientation. RNA was extracted from each treatment and subjected to real time reverse transcriptase-polymerase chain reaction to quantify change in mRNA levels of type I collagen, type III collagen, fibronectin, and TGF-beta1.

RESULT(S): Type I collagen, type III collagen, fibronectin, and TGF-beta1 mRNAs were present in both normal peritoneal and adhesion fibroblasts with significantly higher levels in adhesion fibroblasts. Normal fibroblasts transfected with the COX-2 sense virus exhibited an elevated mRNA level in those molecules that reached the mRNA levels seen for adhesion fibroblasts. There were no effects seen on the mRNA levels in those molecules when adhesion fibroblasts were transfected with COX-2 sense virus. Normal fibroblasts transfected with COX-2 antisense virus exhibited no difference in mRNA levels in those molecules as compared with untransfected controls. In contrast, adhesion fibroblasts transfected with COX-2 antisense exhibited markedly reduced mRNA levels in those molecules to levels that were seen in normal peritoneal fibroblasts.

CONCLUSION(S): Our data suggest that inhibition of COX-2 may reduce the development of postoperative adhesions by preventing the formation of the adhesion phenotype.

摘要

目的

确定阻断环氧合酶-2(COX-2)信使核糖核酸(mRNA)的翻译对从正常腹膜组织和粘连组织获取的成纤维细胞中Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、纤连蛋白和转化生长因子-β1(TGF-β1)mRNA水平的影响。

设计

前瞻性实验研究。

地点

大学医学中心。

患者

从同一患者的腹膜和粘连组织中分离培养的成纤维细胞。

干预措施

用携带COX-2的腺病毒处理原代培养的成纤维细胞。

主要观察指标

从同一患者分离出正常腹膜组织和粘连组织的成纤维细胞。用正义或反义方向的腺病毒COX-2 mRNA转染粘连组织和成纤维细胞。从每种处理中提取RNA,进行实时逆转录聚合酶链反应,以量化Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、纤连蛋白和TGF-β1 mRNA水平的变化。

结果

正常腹膜组织和成纤维细胞中均存在Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、纤连蛋白和TGF-β1 mRNA,粘连组织中的成纤维细胞水平显著更高。用COX-2正义病毒转染的正常成纤维细胞在这些分子中的mRNA水平升高,达到粘连组织中成纤维细胞的mRNA水平。当用COX-2正义病毒转染粘连组织中的成纤维细胞时,这些分子中的mRNA水平未受影响。用COX-2反义病毒转染的正常成纤维细胞在这些分子中的mRNA水平与未转染的对照相比无差异。相比之下,用COX-2反义转染的粘连组织中的成纤维细胞在这些分子中的mRNA水平显著降低,降至正常腹膜组织中成纤维细胞的水平。

结论

我们的数据表明,抑制COX-2可能通过阻止粘连表型的形成来减少术后粘连的发生。

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