N-辛酰多巴胺通过平滑肌细胞保护机制减轻大鼠主动脉同种异体移植血管病变的发展。

N-octanoyl Dopamine Attenuates the Development of Transplant Vasculopathy in Rat Aortic Allografts Via Smooth Muscle Cell Protective Mechanisms.

作者信息

Wedel Johannes, Hottenrott Maximilia C, Bulthuis Marian, Huitema Sippie, Yard Benito A, Hillebrands Jan-Luuk

机构信息

1 Department of Pathology and Medical Biology, Pathology Section, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. 2 Department of Cardiothoracic Surgery, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. 3 Department of Medicine, Nephrology, Endocrinology, Diabetology, Rheumatology, Universitätsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.

出版信息

Transplantation. 2016 Jan;100(1):80-90. doi: 10.1097/TP.0000000000000870.

DOI:10.1097/TP.0000000000000870

PMID:26674731

Abstract

BACKGROUND

Transplant vasculopathy (TV) is a major cause for late graft loss after cardiac transplantation. Endothelial damage and T cell infiltration play a pivotal role in the development of TV. Because N-octanoyl dopamine (NOD) inhibits vascular inflammation and suppresses T cell activation in vitro, we here tested the hypothesis that NOD treatment ameliorates TV.

METHODS

Aortic grafts were orthotopically transplanted in the Dark Agouti to Brown Norway strain combination. Recipient rats were treated with NOD or vehicle administered via osmotic minipumps. Histology and quantitative polymerase chain reaction (qPCR) were performed on nontransplanted aortas and grafts explanted 2 and 4 weeks after transplantation to assess the degree of TV, inflammation, apoptosis, and number of (proliferating) α smooth muscle actin (αSMA) neointimal cells. In vitro analyses of human aortic smooth muscle cells were performed to test the effect of NOD on proliferation (WST-1 assay), cell cycle (flow cytometry and qPCR), and cytokine-induced apoptosis (flow cytometry).

RESULTS

Allografts from vehicle-treated recipients developed neointimal lesions predominantly consisting of αSMA-expressing cells. NOD treatment significantly reduced neointima formation and neointimal αSMA cells. In situ, smooth muscle cell proliferation (Ki67) was not influenced by NOD. Macrophage (CD68), T (CD3), and Natural Killer (ANK61) cell infiltration as well as intragraft TNFα and IFNγ mRNA expression were similar in both groups. Medial apoptosis (cleaved caspase-3) was significantly reduced by NOD. In vitro, NOD inhibited proliferation of human aortic smooth muscle cells by causing a G1-arrest and protected from TNFα-induced apoptosis.

CONCLUSIONS

This study identified NOD as potential treatment modality to attenuate TV. Our data clearly support a vasculoprotective effect of NOD by reducing smooth muscle cell proliferation and inflammation-induced apoptosis.

摘要

背景

移植血管病变（TV）是心脏移植后晚期移植物丢失的主要原因。内皮损伤和T细胞浸润在TV的发展中起关键作用。由于N-辛酰多巴胺（NOD）在体外可抑制血管炎症并抑制T细胞活化，我们在此测试了NOD治疗可改善TV的假说。

方法

将主动脉移植物原位移植到暗褐鼠与棕色挪威大鼠品系组合中。通过渗透微型泵给受体大鼠注射NOD或赋形剂。在移植后2周和4周，对未移植的主动脉和取出的移植物进行组织学检查和定量聚合酶链反应（qPCR），以评估TV的程度、炎症、细胞凋亡以及（增殖的）α平滑肌肌动蛋白（αSMA）新内膜细胞的数量。对人主动脉平滑肌细胞进行体外分析，以测试NOD对增殖（WST-1试验）、细胞周期（流式细胞术和qPCR）以及细胞因子诱导的细胞凋亡（流式细胞术）的影响。

结果

接受赋形剂治疗的受体的同种异体移植物形成了主要由表达αSMA的细胞组成的新内膜病变。NOD治疗显著减少了新内膜形成和新内膜αSMA细胞。在原位，平滑肌细胞增殖（Ki67）不受NOD影响。两组的巨噬细胞（CD68）、T细胞（CD3）和自然杀伤细胞（ANK61）浸润以及移植物内TNFα和IFNγ mRNA表达相似。NOD显著减少了中膜细胞凋亡（裂解的半胱天冬酶-3）。在体外，NOD通过导致G1期阻滞抑制人主动脉平滑肌细胞增殖，并保护细胞免受TNFα诱导的细胞凋亡。