丹皮酚对L-精氨酸诱导的小鼠急性胰腺炎氧化损伤的影响

Effects of Paeonol on the Oxidative Damage of Acute Pancreatitis in Mice Induced by L-Arginine

为了探究丹皮酚对急性胰腺炎(acute pancreatitis,AP)小鼠氧化损伤的影响,本试验选取50只SPF级雄性小鼠,随机分成5组,每组10只,分别为空白对照组、AP模型组和丹皮酚高、中、低剂量组。丹皮酚组小鼠分别灌胃100、50和25 mg·kg^(-1)丹皮酚,同时空白对照组和AP模型组小鼠给予等体积生理盐水。连续灌胃5 d后,采用20%L-精氨酸腹腔注射AP模型组和不同剂量丹皮酚组小鼠,6 h后采集各组小鼠血清测定氧化指标(MDA、SOD),取小鼠胰腺组织肉眼观察病理学变化,并制作胰腺组织髓过氧化物酶(myeloperoxidase,MPO)免疫组化切片,观察其组织病理学变化和MPO的分布及表达。采用荧光定量PCR检测各组胰腺组织中HO-1、Keap1以及Nrf2 mRNA表达水平,采用Western blot检测HO-1、Keap1以及Nrf2蛋白表达量。结果发现,不同剂量的丹皮酚极显著降低急性胰腺炎氧化损伤下小鼠机体的MDA含量,并极显著提高抗氧化酶SOD活力(P<0.01);免疫组化结果显示,高、中剂量丹皮酚有效减少MPO表达;与AP模型组相比,中、高剂量组丹皮酚极显著升高HO-1和Nrf2 mRNA及蛋白表达(P<0.01),低剂量组丹皮酚极显著提高Nrf2 mRNA及蛋白表达(P<0.01),不同剂量组丹皮酚极显著降低Keap1 mRNA和蛋白表达(P<0.01)。综上,丹皮酚可能通过激活Nrf2/HO-1信号通路缓解L-精氨酸诱导的小鼠急性胰腺炎氧化损伤,研究结果为急性胰腺炎发病机制及相关药物的研发提供理论参考。

To investigate the effect of paeonol on oxidative damage in mice with acute pancreatitis(AP),50 SPF male mice were selected and randomly divided into 5 groups with 10 each,namely the blank control group,AP model group and paeonol high-,medium-and low-dose groups.The mice in the paeonol groups were given 100,50,and 25 mg·kg^(-1) paeonol intragastrically,and the mice in the blank control group and the AP model group were given equal volumes of saline.After continuously intragastric administration for 5 days,mice in the AP model group and different doses of paeonol groups were injected intraperitoneally with 20%L-arginine.After 6 hours of injection,serum samples of mice were taken to measure the oxidation indexes(MDA,SOD),and pancreatic tissues of mice were taken to observe the pathological changes.At the same time,myeloperoxidase(MPO)immunohistochemical sections of pancreatic tissue were made to observe the histopathological changes and the distribution and expression of MPO.The mRNA expression levels of HO-1,Keap1 and Nrf2 in pancreatic tissues of each group were detected by fluorescence quantitative PCR,and the protein expression levels of HO-1,Keap1 and Nrf2 were detected by Western blot.The results showed that different doses of paeonol significantly reduced the MDA content and increased the activity of SOD in mice with acute pancreatitis(P<0.01).Immunohistochemical results showed that high and medium dose of paeonol effectively reduced the expression of MPO.Compared with the model group,medium and high dose of paeonol significantly increased the mRNA and protein expression of HO-1 and Nrf2(P<0.01),low dose of paeonol significantly increased the mRNA and protein expression of Nrf2(P<0.01).Different doses of paeonol significantly decreased the expression of Keap1 mRNA and protein(P<0.01).In conclusion,paeonol may alleviate oxidative damage induced by L-arginine in mice with acute pancreatitis by activating Nrf2/HO-1 signaling pathway.This study provides theoretical reference for the pathogenesis of acute pancreatitis and the development of related drugs.