NADPH oxidase plays a crucial role in the activation of pancreatic stellate cells
NADPH oxidase plays a crucial role in the activation of pancreatic stellate cells
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DOI：
10.1152/ajpgi.00272.2007
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发表时间：
2008-01-01
影响因子：
4.5
通讯作者：
Shimosegawa, Tooru
中科院分区：
文献类型：
--
作者：
Masamune, Atsushi;Watanabe, Takashi;Shimosegawa, Tooru
活化的胰腺星状细胞（PSC）在胰腺纤维化和炎症中起重要作用，其中氧化应激参与发病机制。NADPH氧化酶可能是胰腺损伤后活性氧的来源之一。本研究旨在阐明NADPH氧化酶在PSC中的表达及其对细胞功能的调控。从大鼠和人胰腺组织中分离PSC。通过逆转录-PCR和免疫染色评估NADPH氧化酶的表达。使用2 '，7'-二氯荧光素二乙酸酯评估细胞内ROS产生。在体外研究了NADPH氧化酶抑制剂二苯基碘鎓（DPI）和夹竹桃苷对PSC激活关键参数的影响。在体内，DPI（在lmg.对10周龄雄性Wistar Bonn/Kobori大鼠和二丁基二氯化锡（DBTC）诱发的慢性胰腺炎大鼠，饮水给予10 wk（kg体重·天）。PSC表达NADPH氧化酶的关键组分（p22（phox）、p47（phox）、NOX 1、gp 91（phox）/NOX 2、NOX 4和NOX激活剂1）。PDGF-BB、IL-1 β和血管紧张素II诱导ROS产生，DPI和夹竹桃麻素可抑制ROS产生。DPI抑制PDGF诱导的增殖、IL-1 β诱导的趋化因子产生以及α-平滑肌肌动蛋白和胶原蛋白的表达。DPI抑制新鲜分离的细胞转化成肌纤维母细胞样表型。此外，DPI抑制Wistar Bonn/Kobori大鼠和DBTC诱导的慢性胰腺炎大鼠的胰腺纤维化发展。总之，PSC表达NADPH氧化酶以产生ROS，其介导关键细胞功能和PSC的活化。NADPH氧化酶可能是治疗胰腺纤维化的潜在靶点。
Activated pancreatic stellate cells (PSCs) play an important role in pancreatic fibrosis and inflammation, where oxidative stress is implicated in the pathogenesis. NADPH oxidase might be a source of reactive oxygen species (ROS) in the injured pancreas. This study aimed to clarify the expression and regulation of cell functions by NADPH oxidase in PSCs. PSCs were isolated from rat and human pancreas tissues. Expression of NADPH oxidase was assessed by reverse transcription-PCR and immunostaining. Intracellular ROS production was assessed using 2', 7'-dichlorofluorescin diacetate. The effects of diphenylene iodonium (DPI) and apocynin, inhibitors of NADPH oxidase, on key parameters of PSC activation were evaluated in vitro. In vivo, DPI (at 1 mg . kg body wt(-1).day(-1)) was administered in drinking water to 10-wk-old male Wistar Bonn/Kobori rats for 10 wk and to rats with chronic pancreatitis induced by dibutyltin dichloride (DBTC). PSCs expressed key components of NADPH oxidase (p22(phox), p47(phox), NOX1, gp91(phox)/ NOX2, NOX4, and NOX activator 1). PDGF-BB, IL-1 beta, and angiotensin II induced ROS production, which was abolished by DPI and apocynin. DPI inhibited PDGF-induced proliferation, IL-1 beta- induced chemokine production, and expression of alpha-smooth muscle actin and collagen. DPI inhibited transformation of freshly isolated cells to a myofibroblast-like phenotype. In addition, DPI inhibited the development of pancreatic fibrosis in Wistar Bonn/Kobori rats and in rats with DBTC-induced chronic pancreatitis. In conclusion, PSCs express NADPH oxidase to generate ROS, which mediates key cell functions and activation of PSCs. NADPH oxidase might be a potential target for the treatment of pancreatic fibrosis.