Department of Physiology, Jagiellonian University Medical College, 16 Grzegorzecka Street 31-531 Cracow, Poland.
Department of Physiology, Jagiellonian University Medical College, 16 Grzegorzecka Street 31-531 Cracow, Poland.
Biochem Pharmacol. 2018 Mar;149:131-142. doi: 10.1016/j.bcp.2017.11.020. Epub 2017 Dec 1.
Hydrogen sulfide (HS) and carbon monoxide (CO) exert gastroprotection against acute gastric lesions. We determined the cross-talk between HS and CO in gastric ulcer healing process and regulation of gastric blood flow (GBF) at ulcer margin. Male Wistar rats with acetic acid-induced gastric ulcers were treated i.g. throughout 9 days with vehicle (control), NaHS (0.1-10 mg/kg) +/- zinc protoporphyrin (ZnPP, 10 mg/kg), d,l-propargylglycine (PAG, 30 mg/kg), CO-releasing CORM-2 (2.5 mg/kg) +/- PAG. GBF was assessed by laser flowmetry, ulcer area was determined by planimetry/histology. Gastric mucosal HS production was analysed spectrophotometrically. Protein and/or mRNA expression at ulcer margin for vascular endothelial growth factor (VEGF)A, epidermal growth factor receptor (EGFr), cystathionine-γ-lyase (CSE), cystathionine-β-synthase (CBS), 3-mercaptopyruvate sulfurtransferase (3-MST), heme oxygenases (HOs), nuclear factor (erythroid-derived 2)-like 2 (Nrf-2), cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), IL-1β, TNF-α and hypoxia inducible factor (HIF)-1α were determined by real-time PCR or western blot. IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IFN-γ, TNF-α, GM-CSF plasma concentration was assessed using Luminex platform. NaHS dose-dependently decreased ulcer area and increased GBF but ZnPP attenuated these effects. PAG decreased HS production but failed to affect CORM-2-mediated ulcer healing and vasodilation. NaHS increased Nrf-2, EGFr, VEGFA and decreased pro-inflammatory markers expression and IL-1β, IL-2, IL-13, TNF-α, GM-CSF plasma concentration. CORM-2 decreased IL-1β and GM-CSF plasma levels. We conclude that NaHS accelerates gastric ulcer healing increasing microcirculation and Nrf-2, EGFr, VEGFA expression. HS-mediated ulcer healing involves endogenous CO activity while CO does not require HS. NaHS decreases systemic inflammation more effectively than CORM-2.
硫化氢 (HS) 和一氧化碳 (CO) 对急性胃损伤具有胃保护作用。我们确定了 HS 和 CO 在胃溃疡愈合过程中的相互作用以及溃疡边缘胃血流 (GBF) 的调节。用乙酸诱导胃溃疡的雄性 Wistar 大鼠通过口服给予载体 (对照)、NaHS (0.1-10mg/kg) +/- 锌原卟啉 (ZnPP，10mg/kg)、d,l-炔丙基甘氨酸 (PAG，30mg/kg)、CO 释放 CORM-2 (2.5mg/kg) +/- PAG，持续 9 天。通过激光血流仪评估 GBF，通过平面图/组织学确定溃疡面积。用分光光度法分析胃黏膜 HS 生成。通过实时 PCR 或 Western blot 测定溃疡边缘血管内皮生长因子 (VEGF)A、表皮生长因子受体 (EGFr)、胱硫醚-γ-裂解酶 (CSE)、胱硫醚-β-合酶 (CBS)、3-巯基丙酮酸硫转移酶 (3-MST)、血红素加氧酶 (HOs)、核因子 (红系衍生 2)-样 2 (Nrf-2)、环氧化酶 (COX)-2、诱导型一氧化氮合酶 (iNOS)、白细胞介素-1β (IL-1β)、肿瘤坏死因子-α (TNF-α) 和缺氧诱导因子 (HIF)-1α 的蛋白和/或 mRNA 表达。使用 Luminex 平台评估白细胞介素-1α (IL-1α)、白细胞介素-1β (IL-1β)、白细胞介素-2 (IL-2)、白细胞介素-4 (IL-4)、白细胞介素-5 (IL-5)、白细胞介素-6 (IL-6)、白细胞介素-10 (IL-10)、白细胞介素-12 (IL-12)、白细胞介素-13 (IL-13)、干扰素-γ (IFN-γ)、肿瘤坏死因子-α (TNF-α)、粒细胞-巨噬细胞集落刺激因子 (GM-CSF) 血浆浓度。NaHS 剂量依赖性地减少溃疡面积并增加 GBF，但 ZnPP 减弱了这些作用。PAG 减少 HS 生成，但未能影响 CORM-2 介导的溃疡愈合和血管扩张。NaHS 增加 Nrf-2、EGFr、VEGFA 并降低促炎标志物的表达和白细胞介素-1β、白细胞介素-2 (IL-2)、白细胞介素-13 (IL-13)、肿瘤坏死因子-α (TNF-α)、GM-CSF 血浆浓度。CORM-2 降低白细胞介素-1β 和 GM-CSF 血浆水平。我们得出结论，NaHS 通过增加微循环和 Nrf-2、EGFr、VEGFA 的表达来加速胃溃疡愈合。HS 介导的溃疡愈合涉及内源性 CO 活性，而 CO 不需要 HS。NaHS 比 CORM-2 更有效地降低全身炎症。
