Department of Medicine, Mayo Clinic, Scottsdale, Arizona.
Department of Lab Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.
Am J Pathol. 2019 Jun;189(6):1226-1240. doi: 10.1016/j.ajpath.2019.02.015. Epub 2019 Apr 5.
Acute lipolysis of visceral fat or circulating triglycerides may worsen acute pancreatitis (AP)-associated local and systemic injury. The pancreas expresses pancreatic triacylglycerol lipase (PNLIP), pancreatic lipase-related protein 2 (PNLIPRP2), and carboxyl ester lipase (CEL), which may leak into the visceral fat or systemic circulation during pancreatitis. We, thus, aimed to determine the pancreatic lipase(s) regulating lipotoxicity during AP. For this AP, associated fat necrosis was analyzed using Western blot analysis. Bile acid (using liquid chromatography-tandem mass spectrometry) and fatty acid (using gas chromatography) concentrations were measured in human fat necrosis. The fat necrosis milieu was simulated in vitro using glyceryl trilinoleate because linoleic acid is increased in fat necrosis. Bile acid requirements to effectively hydrolyze glyceryl trilinoleate were studied using exogenous or overexpressed lipases. The renal cell line (HEK 293) was used to study lipotoxic injury. Because dual pancreatic lipase knockouts are lethal, exocrine parotid acini lacking lipases were used to verify the results. PNLIP, PNLIPRP2, and CEL were increased in fat necrosis. Although PNLIP and PNLIPRP2 were equipotent in inducing lipolysis and lipotoxic injury, CEL required bile acid concentrations higher than in human fat necrosis. The high bile acid requirements for effective lipolysis make CEL an unlikely mediator of lipotoxic injury in AP. It remains to be explored whether PNLIP or PNLIPRP2 worsens AP severity in vivo.
内脏脂肪或循环中的甘油三酯的急性脂解可能会加重急性胰腺炎(AP)相关的局部和全身损伤。胰腺表达胰腺三酰基甘油脂肪酶(PNLIP)、胰腺脂肪酶相关蛋白 2(PNLIPRP2)和羧基酯脂肪酶(CEL),这些酶在胰腺炎期间可能会渗漏到内脏脂肪或全身循环中。因此,我们旨在确定在 AP 期间调节脂毒性的胰腺脂肪酶。为此,使用 Western blot 分析来分析与 AP 相关的脂肪坏死。使用液相色谱-串联质谱法测量胆汁酸(使用液相色谱-串联质谱法)和脂肪酸(使用气相色谱法)在人脂肪坏死中的浓度。使用甘油三油酸酯模拟脂肪坏死环境,因为在脂肪坏死中油酸增加。使用外源性或过表达的脂肪酶研究了胆汁酸有效水解甘油三油酸酯的要求。使用肾细胞系(HEK 293)研究脂毒性损伤。由于双重胰腺脂肪酶敲除是致命的,因此使用缺乏脂肪酶的外分泌腮腺腺泡来验证结果。在脂肪坏死中增加了 PNLIP、PNLIPRP2 和 CEL。尽管 PNLIP 和 PNLIPRP2 在诱导脂肪分解和脂毒性损伤方面具有同等效力,但 CEL 需要高于人脂肪坏死中的胆汁酸浓度才能发挥作用。有效脂肪分解所需的高胆汁酸要求使 CEL 不太可能成为 AP 中脂毒性损伤的介质。尚待探讨 PNLIP 或 PNLIPRP2 是否会在体内加重 AP 的严重程度。