Hydrogen gas and preservation of intestinal stem cells in mesenteric ischemia and reperfusion.
Hydrogen gas and preservation of intestinal stem cells in mesenteric ischemia and reperfusion.
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DOI：
10.4240/wjgs.v14.i12.1329
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发表时间：
2022-12-27
影响因子：
2
通讯作者：
Sasaki, Junichi
中科院分区：
文献类型：
--
作者：
Yamamoto, Ryo;Suzuki, Sayuri;Homma, Koichiro;Yamaguchi, Shintaro;Sujino, Tomohisa;Sasaki, Junichi
关键词：
肠系膜缺血的患者即使在血运重建后也经常出现肠坏死。在各种动物和临床研究中，氢气通过减少氧自由基而显示出良好的治疗缺血再灌注损伤的效果。我们观察了在3%氢气持续引发下的缺血再灌注对肠组织的损伤。阐明氢气对肠系膜缺血的治疗作用及靶细胞。3组大鼠分别行肠系膜动脉阻断60min(缺血组)、阻断60min后再灌流60min(再灌注组)和持续3%氢气吸入(氢气)60min再灌注组。切取远端回肠。用caspase-3和肠道干细胞特异性标志物富含亮氨酸重复序列的G蛋白偶联5(LGR5)进行免疫荧光染色，评价氢对损伤部位和细胞类型的保护作用。用定量聚合酶链式反应检测LGR5、OLFM4、Split 1的毛发和增强子、Jagge2和Neurogenic Locus Noch同源蛋白1的表达。8-羟基-2‘-脱氧鸟苷(8-OHdG)免疫组织化学染色分析组织氧化应激反应。用血浆8-OHdG评价全身氧化应激反应。缺血损伤绒毛顶端上皮层，再灌流引起隐窝基底部细胞广泛凋亡，免疫荧光双重染色鉴定为肠干细胞。氢气可减轻隐窝底部的这种细胞凋亡，氢气组组织中LGR5的表达高于再灌注组。在氢气组中，Olfm4也相对较高，而其他测量到的RNA在两组之间是相似的。再灌注组8-OHdG浓度较高，氢气可使8-OHdG浓度降低，尤其是在隐窝底部。再灌注组和氢化组的血清8-OHdG浓度均相对较高，但无统计学意义。本研究证明氢气吸入可保护肠干细胞，减轻肠系膜缺血再灌注引起的氧化应激。
Patients with mesenteric ischemia frequently suffer from bowel necrosis even after revascularization. Hydrogen gas has showed promising effects for ischemia-reperfusion injury by reducing reactive oxygen species in various animal and clinical studies. We examined intestinal tissue injury by ischemia and reperfusion under continuous initiation of 3% hydrogen gas. To clarify the treatment effects and target cells of hydrogen gas for mesenteric ischemia. Three rat groups underwent 60-min mesenteric artery occlusion (ischemia), 60-min reperfusion following 60-min occlusion (reperfusion), or ischemia-reperfusion with the same duration under continuous 3% hydrogen gas inhalation (hydrogen). The distal ileum was harvested. Immunofluorescence staining with caspase-3 and leucine-rich repeat-containing G-protein-coupled 5 (LGR5), a specific marker of intestinal stem cell, was conducted to evaluate the injury location and cell types protected by hydrogen. mRNA expressions of LGR5, olfactomedin 4 (OLFM4), hairy and enhancer of split 1, Jagged 2, and Neurogenic locus notch homolog protein 1 were measured by quantitative polymerase chain reaction. Tissue oxidative stress was analyzed with immunostaining for 8-hydroxy-2'-deoxyguanosine (8-OHdG). Systemic oxidative stress was evaluated by plasma 8-OHdG. Ischemia damaged the epithelial layer at the tip of the villi, whereas reperfusion induced extensive apoptosis of the cells at the crypt base, which were identified as intestinal stem cells with double immunofluorescence stain. Hydrogen mitigated such apoptosis at the crypt base, and the LGR5 expression of the tissues was higher in the hydrogen group than in the reperfusion group. OLFM4 was also relatively higher in the hydrogen group, whereas other measured RNAs were comparable between the groups. 8-OHdG concentration was high in the reperfusion group, which was reduced by hydrogen, particularly at the crypt base. Serum 8-OHdG concentrations were relatively higher in both reperfusion and hydrogen groups without significance. This study demonstrated that hydrogen gas inhalation preserves intestinal stem cells and mitigates oxidative stress caused by mesenteric ischemia and reperfusion.
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