Oshitari Toshiyuki, Brown David, Roy Sayon
Department of Medicine, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118, USA.
Exp Eye Res. 2005 Jul;81(1):32-7. doi: 10.1016/j.exer.2005.01.006.
Increased synthesis of extracellular matrix (ECM) contributes to the development of vascular BM thickening, a prominent abnormality in diabetic retinopathy. RNA interference (RNAi) approach was used in this study to examine the effect of small interfering RNAs (siRNAs) for their ability to inhibit ECM-specific gene overexpression under high glucose condition in rat microvascular endothelial cells (RMECs). Four fibronectin (FN)-siRNAs, three collagen IV (Coll IV)-siRNAs, and four laminin (LM)-siRNAs, a total of 11 siRNAs were screened. RMECs were transfected with 10, 30, or 100 nm of each siRNAs in the presence of 8 microm lipofectin and subjected to analysis 72 hr after transfection. In long-term studies siRNA-transfected cells were examined after 12 days. Two FN siRNAs, two Coll IV siRNAs, and two LM siRNAs significantly reduced the respective target expressions. Findings from this study indicate that high glucose-induced abnormal expression of BM components may contribute to increased vascular permeability. SiRNA may be a useful tool in preventing excess vascular permeability, a characteristic feature of early diabetic retinopathy.
细胞外基质（ECM）合成增加促使血管基底膜增厚，这是糖尿病视网膜病变的一个显著异常。本研究采用RNA干扰（RNAi）方法，检测小干扰RNA（siRNA）在高糖条件下抑制大鼠微血管内皮细胞（RMECs）中ECM特异性基因过表达的能力。共筛选了4种纤连蛋白（FN）-siRNA、3种IV型胶原（Coll IV）-siRNA和4种层粘连蛋白（LM）-siRNA，总计11种siRNA。在存在8微摩尔脂质体转染试剂的情况下，将每种siRNA以10、30或100纳米的浓度转染RMECs，并在转染后72小时进行分析。在长期研究中，对转染siRNA的细胞在12天后进行检测。两种FN siRNA、两种Coll IV siRNA和两种LM siRNA显著降低了各自的靶标表达。本研究结果表明，高糖诱导的基底膜成分异常表达可能导致血管通透性增加。SiRNA可能是预防早期糖尿病视网膜病变特征性的血管通透性过高的有用工具。
