Paulino Ellena C, de Souza Lourenilson J, Molan Nilza A T, Machado Marcel C C, Jancar Sonia
Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, Brazil.
Pancreas. 2007 Jul;35(1):37-41. doi: 10.1097/MPA.0b013e31805c177b.
There is evidence that endothelin (ET) 1 affect neutrophil functions and that patients with severe acute pancreatitis have increased plasma levels of ETs. Under appropriate conditions, neutrophils are able to injure the endothelium. In the present study, we compared healthy donors with acute pancreatitis patients for neutrophil degranulation and its ability to injure the endothelium and the contribution of ET-1 to this injury.
Injury was evaluated by measuring the detachment of endothelial cells (ECV-304) growing in monolayer in coculture with human neutrophils for 4 hours. Neutrophil degranulation was assessed by myeloperoxidase (MPO) activity in coculture supernatants. In some experiments, neutrophils were pretreated with the antagonist of ET(A) receptor (BQ-123, 10(-6) M), which has high affinity for ET-1.
Neutrophils from both healthy donors and acute pancreatitis patients caused detachment of endothelial cells, and levels of MPO activity were increased in coculture supernatants. Neutrophils from acute pancreatitis patients caused significantly higher levels of detachment and MPO in the supernatants. Pretreatment of neutrophils with BQ-123 inhibited the detachment caused by neutrophils from healthy donors but not by neutrophils from acute pancreatitis patients.
These results show that neutrophils taken from healthy donors damage the endothelium by a mechanism dependent on ETs acting via ET(A) receptor, whereas neutrophils from acute pancreatitis patients cause more severe damage that is not dependent on ETs in the in vitro system used.
有证据表明内皮素（ET）-1影响中性粒细胞功能，且重症急性胰腺炎患者血浆中ET水平升高。在适当条件下，中性粒细胞能够损伤内皮。在本研究中，我们比较了健康供体和急性胰腺炎患者中性粒细胞的脱颗粒情况、其损伤内皮的能力以及ET-1对这种损伤的作用。
通过测量与人类中性粒细胞共培养4小时的单层生长内皮细胞（ECV-304）的脱离情况来评估损伤。通过共培养上清液中的髓过氧化物酶（MPO）活性评估中性粒细胞脱颗粒情况。在一些实验中，中性粒细胞用ET(A)受体拮抗剂（BQ-123，10(-6) M）预处理，该拮抗剂对ET-1具有高亲和力。
健康供体和急性胰腺炎患者的中性粒细胞均导致内皮细胞脱离，且共培养上清液中MPO活性水平升高。急性胰腺炎患者的中性粒细胞导致的脱离水平和上清液中的MPO水平显著更高。用BQ-123预处理中性粒细胞可抑制健康供体中性粒细胞引起的脱离，但不能抑制急性胰腺炎患者中性粒细胞引起的脱离。
这些结果表明，从健康供体获取的中性粒细胞通过依赖ET通过ET(A)受体发挥作用的机制损伤内皮，而在所用的体外系统中，急性胰腺炎患者的中性粒细胞造成的损伤更严重，且不依赖ET。
