Laboratory of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
Biol Pharm Bull. 2010;33(2):307-10. doi: 10.1248/bpb.33.307.
Tribenoside has been used clinically for hemorrhoidal disease associated with coagulation, inflammation, and wounds. However, the pharmacological mechanism of tribenoside activity has never been clear. In this study we examined whether tribenoside affected expression and deposition of laminins that are required for reconstruction of basement membranes (BMs) during wound healing in hemorrhoidal disease. HaCaT cells, which are derived from human epidermis, were treated in growth media supplemented with tribenoside. Reverse transcriptase-polymerase chain reaction (RT-PCR) using primers specific for laminin chains showed that HaCaT cells constitutively expressed laminin alpha3, alpha5, beta1, beta3, gamma1, and gamma2 chains. Tribenoside treatment of HaCaT cells did not induce expression of other laminin chains. We also quantified the expression of laminin chains in tribenoside-treated cells using real-time PCR. The expression level of laminin alpha3, beta1, beta3, gamma1, and gamma2 chains was not affected. In contrast, the expression of laminin alpha5 in the tribenoside-treated cells was four times higher than that of control cells. Immunocytochemistry also showed that tribenoside accelerated the focal deposition of laminin-332 (alpha3, beta3, gamma2). These results suggest that tribenoside interacts with epidermal cells and regulates the expression and localization of laminins to help reconstruct BMs in wound healing of hemorrhoids.
地奥司明临床上用于治疗与凝血、炎症和创伤相关的痔疮疾病。然而，地奥司明的药理机制尚不清楚。在这项研究中，我们研究了地奥司明是否影响层粘连蛋白的表达和沉积，层粘连蛋白是在痔疮疾病的伤口愈合过程中重建基底膜（BMs）所必需的。HaCaT 细胞来源于人表皮，在补充有地奥司明的生长培养基中进行处理。使用针对层粘连蛋白链的特异性引物的逆转录-聚合酶链反应（RT-PCR）显示，HaCaT 细胞持续表达层粘连蛋白 alpha3、alpha5、beta1、beta3、gamma1 和 gamma2 链。地奥司明处理 HaCaT 细胞不会诱导其他层粘连蛋白链的表达。我们还使用实时 PCR 定量测定了地奥司明处理细胞中层粘连蛋白链的表达。层粘连蛋白 alpha3、beta1、beta3、gamma1 和 gamma2 链的表达水平不受影响。相比之下，地奥司明处理细胞中层粘连蛋白 alpha5 的表达是对照细胞的四倍。免疫细胞化学也表明，地奥司明加速了层粘连蛋白-332（alpha3、beta3、gamma2）的焦点沉积。这些结果表明，地奥司明与表皮细胞相互作用，调节层粘连蛋白的表达和定位，以帮助重建痔疮伤口愈合中的 BM。
