HO-1在重症急性胰腺炎引起的肠黏膜损伤中的作用及其机制

Effects and Mechanism of HO-1 in Intestinal Mucosal Injury Caused by Severe Acute Pancreatitis

摘要: 目的：本研究旨在探究HO-1对SAP大鼠肠黏膜的保护作用及机制。方法：选取SPF级健康雄性SD大鼠32只，随机分为假手术组(SO组)、重症急性胰腺炎组(SAP组)、HO-1激活组(SAP + Hemin组)、HO-1抑制组(SAP + Znpp组)，每组8只。采用5%牛磺胆酸钠逆行注入胆胰管的方法建立大鼠SAP模型，SO组胆胰管注入等量生理盐水。干预组大鼠分别在建模前24 h腹腔注射Hemin和Znpp，其余组腹腔注射等量生理盐水。各组大鼠在手术后24 h检测大鼠血清淀粉酶、TNF-α、IL-6、IL-10、DAO活性以及D-LA、FABP2和内毒素水平；检测回肠组织MDA含量以及SOD和GPX的活性水平；苏木精–伊红(HE)染色观察胰腺、回肠病理变化；免疫荧光染色检测HO-1及Occludin、ZO-1的表达情况；TUNEL检测肠上皮细胞凋亡水平。结果：SAP组大鼠血清AMY、TNF-α、IL-6、DAO活性以及D-LA、FABP2和内毒素水平明显高于SO组，IL-10显著低于SO组(均P < 0.05)。与SAP组相比，SAP + Hemin组显著增加HO-1的表达，减轻了SAP全身炎症反应、MDA含量以及SOD和GPX的活性明显降低，肠上皮细胞凋亡减少(均P < 0.05)。同时，上调HO-1的表达减轻了对肠紧密连接蛋白表达的抑制作用，血清DAO活性及D-LA、FABP2和内毒素水平显著降低(均P < 0.05)。相反，SAP + Znpp组则表现出相反的实验结果。结论：H0-1通过减轻炎症、氧化应激及抗凋亡的作用来减轻重症急性胰腺炎肠粘膜损伤。

Abstract: Objective: To investigate the protective effect and mechanism of HO-1 on intestinal mucosa in SAP rats. Methods: Thirty-two SPF-grade healthy male SD rats were selected and randomly divided into sham-operated group (SO group), severe acute pancreatitis group (SAP group), HO-1 activation group (SAP + Hemin group), and HO-1 inhibition group (SAP + Znpp group), eight rats in each group. The rat SAP model was established through retrograde injection of 5% sodium taurocholate into the biliopancreatic duct, and the SO group was injected with equal amounts of saline into the bili-opancreatic duct. Hemin and Znpp were injected intraperitoneally 24 h before modeling in the in-tervention group, while equal amounts of saline were injected intraperitoneally in the other group. At 24 hours after surgery, serum amylase, TNF-α, IL-6, IL-10, DAO, D-LA, FABP2 and endotoxin lev-els were measured in rats; the MDA content and the activity levels of SOD and GPX in ileal tissue were measured; Hematoxylin and eosin (HE) staining for pathological changes in the pancreas and ileum; Immunofluorescence staining detection of HO-1 and Occludin, ZO-1 expression; TUNEL de-tection of apoptosis level in intestinal epithelial cells. Results: The serum AMY, TNF-α, IL-6, DAO, D-LA, FABP2 and endotoxin levels of rats in SAP group were significantly higher than those in SO group, and IL-10 was significantly lower than that in SO group (all P < 0.05). Compared with the SAP group, the SAP + Hemin group significantly increased HO-1 expression, attenuated systemic in-flammatory response, MDA content, and significantly reduced SOD and GPX activities, and de-creased intestinal epithelial cell apoptosis (all P < 0.05). Meanwhile, upregulation of HO-1 expres-sion attenuated the inhibitory effect on intestinal tight junction protein expression, and serum DAO, D-LA, FABP2 and endotoxin levels were significantly reduced (all P < 0.05). However, the SAP + Znpp group showed the opposite experimental results. Conclusion: HO-1 attenuates intestinal mu-cosal injury in severe acute pancreatitis by reducing inflammation, oxidative stress and an-ti-apoptotic effects.