Susomboon Pannapa, Maneerat Yaowapa, Dekumyoy Paron, Kalambaheti Thareerat, Iwagami Moritoshi, Komaki-Yasuda Kanako, Kawazu Shin-Ichiro, Tangpukdee Noppadon, Looareesuwan Sornchai, Kano Shigeyuki
Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchavithi Road, Bangkok 10400, Thailand.
Parasitol Int. 2006 Jun;55(2):107-12. doi: 10.1016/j.parint.2005.11.054. Epub 2006 Jan 4.
To understand the mechanism of sequestration in the microvasculature of patients with falciparum malaria, we examined the patterns of expression of mRNAs for adhesion molecules (ICAM-1, VCAM-1, and E-selectin) and tight junction molecules (occludin, vinculin, and ZO-1) in human umbilical vein endothelial cells (HUVECs) co-cultured with Plasmodium falciparum-parasitized red blood cells (PRBCs) in vitro. The PRBCs were collected from patients with uncomplicated, severe, or cerebral malaria (CM). Patterns of mRNA expression in HUVECs co-cultured with PRBCs were examined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Levels of mRNAs for all the three adhesion molecules increased with increased culture time within 3 h, regardless of the source of the PRBCs. In contrast, the patterns of mRNA expression for the tight junction molecules varied between the different co-cultures. When HUVECs were cultured with PRBCs from uncomplicated malaria patients, levels of mRNAs for tight junction molecules increased according to the culture time. HUVECs co-cultured with PRBCs from severe malaria patients showed no change in the mRNAs levels during 3 h of observation. When HUVECs were cultured with PRBCs from CM patients, levels of mRNAs for tight junction proteins decreased according to the culture time. Although the mechanisms underlying these phenomena are not clear, our results suggest that PRBCs can alter expression of tight junction proteins in endothelial cells at the site of sequestration and thereby influence disease severity.
为了解恶性疟患者微血管中隔离的机制，我们检测了体外与人恶性疟原虫感染的红细胞（PRBCs）共培养的人脐静脉内皮细胞（HUVECs）中黏附分子（ICAM-1、VCAM-1和E-选择素）和紧密连接分子（闭合蛋白、纽蛋白和ZO-1）的mRNA表达模式。PRBCs取自非重症、重症或脑型疟（CM）患者。通过实时定量逆转录聚合酶链反应（RT-PCR）检测与PRBCs共培养的HUVECs中的mRNA表达模式。在3小时内，所有三种黏附分子的mRNA水平均随培养时间增加而升高，与PRBCs的来源无关。相比之下，不同共培养组中紧密连接分子的mRNA表达模式有所不同。当HUVECs与非重症疟疾患者的PRBCs共培养时，紧密连接分子的mRNA水平随培养时间增加。与重症疟疾患者的PRBCs共培养的HUVECs在观察的3小时内mRNA水平无变化。当HUVECs与CM患者的PRBCs共培养时，紧密连接蛋白的mRNA水平随培养时间下降。尽管这些现象背后的机制尚不清楚，但我们的结果表明，PRBCs可改变隔离部位内皮细胞中紧密连接蛋白的表达，从而影响疾病严重程度。
