Ventimiglia María Silvia, Najenson Ana Clara, Perazzo Juan Carlos, Carozzo Alejandro, Vatta Marcelo S, Davio Carlos A, Bianciotti Liliana G
Cátedra de Fisiopatología, Instituto de Inmunología, Genética y Metabolismo (INIGEM-CONICET), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
Laboratorio de Farmacología de Receptores, Cátedra de Química Medicinal, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
Mol Med. 2015 Jan 6;21(1):58-67. doi: 10.2119/molmed.2014.00166.
We previously reported that atrial natriuretic factor (ANF) stimulates secretin-evoked cAMP efflux through multidrug resistance-associated protein 4 (MRP4) in the exocrine pancreas. Here we sought to establish in vivo whether this mechanism was involved in acute pancreatitis onset in the rat. Rats pretreated with or without probenecid (MRPs general inhibitor) were infused with secretin alone or with ANF. A set of these animals were given repetitive cerulein injections to induce acute pancreatitis. Plasma amylase and intrapancreatic trypsin activities were measured and histological examination of the pancreas performed. Secretin alone activated trypsinogen but induced no pancreatic histological changes. Blockade by probenecid in secretin-treated rats increased trypsin and also induced vacuolization, a hallmark of acute pancreatitis. ANF prevented the secretin response but in the absence of probenecid. In rats with acute pancreatitis, pretreatment with secretin aggravated the disease, but ANF prevented secretin-induced changes. Blockade of MRPs in rats with acute pancreatitis induced trypsinogen activation and larger cytoplasmic vacuoles as well as larger areas of necrosis and edema that were aggravated by secretin but not prevented by ANF. The temporal resolution of intracellular cAMP levels seems critical in the onset of acute pancreatitis, since secretin-evoked cAMP in a context of MRP inhibition makes the pancreas prone to injury in normal rats and aggravates the onset of acute pancreatitis. Present findings support a protective role for ANF mediated by cAMP extrusion through MRP4 and further suggest that the regulation of MRP4 by ANF would be relevant to maintain pancreatic acinar cell homeostasis.
我们之前报道过,心房利钠因子(ANF)通过外分泌胰腺中的多药耐药相关蛋白4(MRP4)刺激促胰液素诱发的cAMP流出。在此,我们试图在体内确定该机制是否参与大鼠急性胰腺炎的发病过程。预先用丙磺舒(MRPs的通用抑制剂)处理或未处理的大鼠,分别单独输注促胰液素或与ANF一起输注。对其中一组动物进行重复的雨蛙素注射以诱导急性胰腺炎。测量血浆淀粉酶和胰腺内胰蛋白酶活性,并对胰腺进行组织学检查。单独的促胰液素可激活胰蛋白酶原,但未引起胰腺组织学变化。丙磺舒对促胰液素处理的大鼠的阻断作用增加了胰蛋白酶水平,还诱导了空泡化,这是急性胰腺炎的一个标志。ANF可预防促胰液素反应,但这是在没有丙磺舒的情况下。在患有急性胰腺炎的大鼠中,用促胰液素预处理会加重病情,但ANF可预防促胰液素诱导的变化。对患有急性胰腺炎的大鼠中的MRPs进行阻断会诱导胰蛋白酶原激活、更大的细胞质空泡以及更大面积的坏死和水肿,促胰液素会加重这些变化,但ANF无法预防。细胞内cAMP水平的时间分辨率在急性胰腺炎的发病过程中似乎至关重要,因为在MRP抑制的情况下,促胰液素诱发的cAMP会使正常大鼠的胰腺易于受损,并加重急性胰腺炎的发病。目前的研究结果支持ANF通过MRP4介导的cAMP外排发挥保护作用,并进一步表明ANF对MRP4的调节与维持胰腺腺泡细胞内稳态相关。