|
|
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
|
Abstract:
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。
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
| [1] | Petrov, M.S., Shanbhag, S., Chakraborty, M., Phillips, A.R. and Windsor, J.A. (2010) Organ Failure and Infection of Pancreatic Necrosis as Determinants of Mortality in Patients with Acute Pancreatitis. Gastroenterology, 139, 813-820.
https://doi.org/10.1053/j.gastro.2010.06.010 |
| [2] | Liu, J.B., Huang, L., Luo, M. and Xia, X.M. (2019) Bacterial Translocation in Acute Pancreatitis. Critical Reviews in Microbiology, 45, 539-547. https://doi.org/10.1080/1040841X.2019.1621795 |
| [3] | Huang, L.Q., Jiang, Y.J., Sun, Z.Q., Gao, Z.Y., Wang, J. and Zhang, D.L. (2018) Autophagy Strengthens Intestinal Mucosal Barrier by Attenuating Oxidative Stress in Severe Acute Pancreatitis. Digestive Diseases and Sciences, 63, 910-919. https://doi.org/10.1007/s10620-018-4962-2 |
| [4] | Ma, D.L., Jiang, P.L., Jiang, Y.J., Li, H.B. and Zhang, D.L. (2021) Effects of Lipid Peroxidation-Mediated Ferroptosis on Severe Acute Pancreatitis-Induced Intestinal Barrier Injury and Bacterial Translocation. Oxidative Medicine and Cellular Longevity, 2021, Article ID: 6644576. https://doi.org/10.1155/2021/6644576 |
| [5] | 田锐, 许飞, 王瑞兰, 谢晖, 孟潇潇, 钱永兵, 金卫, 胡家昌, 周志刚, 俞康龙. 氧化应激及凋亡与重症急性胰腺炎肠屏障功能障碍[J]. 中华急诊医学杂志, 2012, 21(10): 1088-1092. |
| [6] | Ryter, S.W. (2021) Heme Oxgenase-1, a Cardinal Modulator of Regulated Cell Death and Inflamma-tion. Cells, 10, Article No. 515. https://doi.org/10.3390/cells10030515 |
| [7] | 张雁林, 赵金垣. 血红素加氧酶-1的细胞保护作用研究进展[J]. 环境与职业医学, 2008, 25(2): 197-202.
https://doi.org/10.13213/j.cnki.jeom.2008.02.020 |
| [8] | Puentes-Pardo, J.D., Moreno-SanJuan, S., Carazo, á. and León, J. (2020 Heme Oxygenase-1 in Gastrointestinal Tract Health and Disease. Antioxidants, 9, Article No. 1214. https://doi.org/10.3390/antiox9121214 |
| [9] | Zhang, Z.L., Zhang, Q.P., Li, F., Xin, Y. and Duan, Z.J. (2021) Con-tributions of HO-1-Dependent MAPK to Regulating Intestinal Barrier Disruption. Biomolecules & Therapeutics, 29, 175-183.
https://doi.org/10.4062/biomolther.2020.112 |
| [10] | 任萍萍, 段志军. 血红素氧合酶-1在胃肠道疾病中的作用研究进展[J]. 大连医科大学学报, 2022, 44(1): 69-74. |
| [11] | Okumura, R. and Takeda, K. (2017) Roles of Intestinal Epi-thelial Cells in the Maintenance of Gut Homeostasis. Experimental & Molecular Medicine, 49, e338. https://doi.org/10.1038/emm.2017.20 |
| [12] | Murray, M.J., Barbose, J.J. and Cobb, C.F. (1993) Serum D(-)-Lactate Levels as a Predictor of Acute Intestinal Ischemia in a Rat Model. The Journal of Surgical Research, 54, 507-509. https://doi.org/10.1006/jsre.1993.1078 |
| [13] | Luk, G.D., Bayless, T.M. and Baylin, S.B. (1980) Diamine Oxidase (Histaminase). A Circulating Marker for Rat Intestinal Mucosal Maturation and Integrity. The Journal of Clinical Inves-tigation, 66, 66-70.
https://doi.org/10.1172/JCI109836 |
| [14] | Funaoka, H., Kanda, T., Kajiura, S., Ohkaru, Y. and Fujii, H. (2011) De-velopment of a High-Specificity Sandwich ELISA System for the Quantification of Human Intestinal Fatty Acid-Binding Protein (I-FABP) Concentrations. Immunological Investigations, 40, 223-242. https://doi.org/10.3109/08820139.2010.534216 |
| [15] | Sun, Z., Wang, X., Wallen, R., Deng, X., Du, X., Hallberg, E. and Andersson, R. (1998) The Influence of Apoptosis on Intestinal Barrier Integrity in Rats. Scandinavian Journal of Gastroenterology, 33, 415-422.
https://doi.org/10.1080/00365529850171053 |
