还原型烟酰胺腺嘌呤二核苷酸磷酸对肠缺血再灌注损伤的影响  

Effect of reduced nicotinamide adenine dinucleotide phosphate on intestinal ischemia-reperfusion injury

作　　者：陈苏颖 徐慧[1] 何天琦 邢雨润 刘以勤 何伯圣[2] 顾锦华[1] CHEN Suying;XU Hui;HE Tianqi;XING Yurun;LIU Yiqin;HE Bosheng;GU Jinhua(Nantong Institute of Genetics and Reproductive Medicine,Affiliated Maternity&Child Healthcare Hospital of Nantong University,Nantong 226007,Jiangsu,China;Department of Radiology,Affiliated Hospital 2 of Nantong University,Medical School of Nantong University,Nantong 226000,Jiangsu,China)

机构地区：[1]南通大学附属妇幼保健院药学部,南通226007 [2]南通大学第二附属医院影像科,南通226000

出　　处：《医学研究与战创伤救治》2025年第2期120-126,共7页Journal of Medical Research & Combat Trauma Care

基　　金：国家科学自然基金(81870941)。

摘　　要：目的研究还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)对小鼠肠缺血再灌注(IR)中的影响及潜在机制。方法将小鼠随机数字表法分为Sham组、IR组、IR+NADPH(10 mg/kg)组和IR+NADPH(20 mg/kg)组。体内构建短暂性肠IR损伤小鼠模型,通过激光散斑血流仪判断模型是否构建成功;通过苏木素-伊红(HE)染色、湿干重比观察肠道黏膜损伤和肠道水肿程度;采用RT-PCR测定炎症因子白细胞介素(IL)-6、IL-1β及肿瘤坏死因子(TNF)-α的表达;通过丙二醛和还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)的水平探索NADPH的抗氧化能力;通过Western blot分析检测Bcl-2关联X蛋白(Bax)、B-细胞淋巴瘤因子(Bcl)-2及半胱氨酸蛋白酶(Cleaved caspase)-3蛋白的表达。结果体内肠IR损伤引起肠道血供显著减少,再灌注后肠道血流再通,肠道组织红肿瘀血。与IR组相比,NADPH加药组显著减轻肠IR小鼠肠道黏膜损伤和肠道水肿程度(P<0.05)。与IR组相比,NADPH加药组可以降低促炎因子IL-6、IL-1β和TNF-α的表达(P<0.05),显著减轻IR损伤后的炎症反应。NADPH加药组有效降低肠道中的丙二醛水平,增高GSH/GSSG比值(均P<0.05),显著减轻损伤后的氧化应激。NADPH加药组可以增加抗凋亡蛋白Bcl-2的表达,降低促凋亡蛋白Bax及Cleaved caspase-3的表达,显著减轻IR损伤后的凋亡水平(P<0.05)。结论NADPH可以通过减轻氧化应激、炎症水平及凋亡水平,从而发挥保护作用。Objective To investigate the effects of reduced nicotinamide adenine dinucleotide phosphate(NADPH)on intestinal ischemia reperfusion(IR)in mice and its potential mechanism.Methods Mice were divided into Sham group,IR group,IR+NADPH(10mg/kg)group and IR+NADPH(20mg/kg)group by random number table method.A mouse model of transient intestinal IR injury was constructed in vivo,and the success of the model was judged by laser speckle blood flow analyzer.The intestinal mucosal injury and intestinal edema were observed by hematoxylin eosin(HE)staining and wet-dry weight ratio.RT-PCR was used to determine the expression of inflammatory cytokines interleukin(IL)6,IL1βand tumor necrosis factor(TNF)α.The antioxidant capacity of NADPH was investigated by the levels of malondialdehyde and reduced glutathione/oxidized glutathione(GSH/GSSG).Western blot analysis was used to detect the expression of Bcl2 associated X protein(Bax),B-cell lymphoma factor(BCL2)and Cleaved caspase 3 protein.Results In vivo intestinal IR injury caused a significant decrease in intestinal blood supply,intestinal blood flow recirculation after reperfusion,and intestinal tissue redness and stasis.Compared with IR group,NADPH group significantly alleviated intestinal mucosal injury and intestinal edema in IR mice(P<0.05).Compared with IR group,NADPH group could decrease the expressions of pro-inflammatory factors IL6,IL1βand TNF-α(P<0.05),and significantly reduce the inflammatory response after IR injury.The levels of malondialdehyde in intestinal tract and the ratio of GSH/GSSG were increased in NADPH group(all P<0.05),and the oxidative stress after injury was significantly alleviated.In NADPH group,the expression of anti-apoptotic protein Bcl2 was in-creased,the expression of pro-apoptotic protein Bax and Cleaved caspase3 were decreased,and the apoptosis level after IR injury was significantly reduced(P<0.05).Conclusion NADPH can play a protective role by alleviating oxidative stress,inflammation level and apoptosis level.

关 键 词：还原型烟酰胺腺嘌呤二核苷酸磷酸 肠缺血再灌注损伤 氧化应激 炎症 凋亡 

分 类 号：R574[医药卫生—消化系统]