NADPH氧化酶-4和酸敏感受体在心理应激小鼠食管中的表达及作用  

The expression of NADPH oxidase-4 and acid sensitive receptors in the esophagus of chronic restraint mice

作　　者：买买提·依斯热依力 阿孜古丽·阿力木江 李义亮[1,2] 艾克拜尔·艾力[1,2] 赵新胜 王志[2] 闫晶[2] 克力木·阿不都热依木[1,2] Maimaiti Yisireyili;Aziguli Alimujiang;Li Yiliang;Aikebaier Aili;Zhao Xinsheng;Wang Zhi;Yan Jing;Kelimu Abudureyimu(Research Institute of General and Minimally Invasive Surgery,People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China;Department of Minimally Invasive Surgery,People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China;Department of Obstetrics and Gynecology Clinic,People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院普外微创研究所,乌鲁木齐830001 [2]新疆维吾尔自治区人民医院微创外科,乌鲁木齐830001 [3]新疆维吾尔自治区人民医院妇产科,乌鲁木齐830001

出　　处：《中国医师杂志》2020年第7期1000-1004,共5页Journal of Chinese Physician

基　　金：新疆维吾尔自治区自然科学基金(2018D01C134)。

摘　　要：目的探讨心理应激诱导酸敏感受体在小鼠食管中的表达及氧化应激在食管炎症发生中的作用.方法选取雄性无特定病原体(SPF)级20只昆明小鼠随机分2组(每组10只),即慢性束缚应激(Stress)组和正常对照(Control)组.应激组小鼠每天在自制式束缚器中限制活动2h,其余时间两组小鼠在相同环境中自由饮水摄食,实验持续14 d.光镜下观察苏木精-伊红(HE)染色后食管黏膜组织病理学改变,采用免疫组化、实时荧光定量PCR(qRT-PCR)、酶联免疫吸附测定方法检测烟酰胺腺嘌呤二核苷酸磷酸氧化酶-4(Nox-4)在小鼠食管及血清中的表达.进一步通过qRT-PCR方法检测食管组织中酸敏感受体的表达水平.结果光镜下观察发现,HE染色后应激组小鼠食管黏膜组织中性粒细胞、嗜酸性粒细胞及浆细胞浸润反应和炎症性改变,而对照组小鼠食管全层未发现明显异常.两组小鼠炎症评分结果显示,应激组小鼠食管黏膜损伤评分均明显高于对照组,差异有统计学意义(P<0.001).免疫组化结果显示,Nox-4主要表达于食管固有层、黏膜及黏膜下层,应激组Nox-4阳性表达显著高于对照组.应激组小鼠食管黏膜组织中Nox-4 mRNA表达水平是对照组的(2.67±0.62)倍,差异有统计学意义(P<0.001);Nox-4在应激组小鼠血清中的表达水平显著高于对照组[(0.42±0.01)ng/ml vs(2.13±0.35)ng/ml],差异具有统计学意义(P<0.001).应激组酸敏感受体如瞬时感受器电位通道-1(TRPV-1)、TRPV-4、酸敏感离子通道-1(ASIC-1)、ASIC-2和ASIC-3的mRNA表达明显高于对照组,差异有统计学意义(P<0.001).Pearson相关性分析结果显示,应激组Nox-4 mRNA表达与TRPV-1、TRPV-4、ASIC-1、ASIC-2、ASIC-3 mRNA表达之间呈正相关(r=0.97、0.94、0.98、0.95、0.99,P<0.01).结论心理应激引起酸敏感受体的异常表达,并诱导炎症和氧化应激产生,从而导致食管高敏感性的产生.Objective To investigate the chronic restraint stress induced expressions of acid sensitive receptors and its role in the esophageal inflammation and oxidative stress.Methods Twenty male specific pathogen free(SPF)Kunming mice were randomly divided into two groups:stress group and control group(each group,n=10).Stress mice were restrained in self-made restraint device for 2 hours per day and lasted for total 14 days.The histopathological changes of esophageal mucosa were observed by hematoxylin eosin(HE)staining under light microscope.The expression of nicotinamide adenine dinucleotide phosphate(Nox-4)was detected by immunohistochemistry,real time fluorescent quantitative polymerase chain reaction(qRT-PCR)and enzyme linked immunosorbent assay(ELISA).The mRNA expressions of acid sensitive receptors were detected by qRT-PCR.Results HE staining showed that stress mice had obvious infiltrations of neutrophils and eosinophils,and also showed inflammatory change in esophgus,while no significant abnormality was found in the esophagus of control mice.The inflammotory scores in stress group were significantly higher than that in control group(P<0.001).Immunohistochemistry showed that Nox-4 was mainly expressed in the lamina propria,mucosa and submucosa of esophagus.The mRNA expression levels of Nox-4 in stress group was(2.67±0.62)times higher than control group,with statistically significant difference(P<0.001).In addition,the plasma concentration of Nox-4 in stress group was significantly higher than that of control group[(0.42±0.01)ng/ml vs(2.13±0.35)ng/ml,P<0.001].The transcription levels of acid sensitive receptors in stressed mice,such as transient receptor potential vanilloid-1(TRPV-1),TRPV-4,acid-sensing ion channel-1(ASIC-1),ASIC-2 and ASIC-3 were significantly higher than those in the control group,with statistically significant difference(P<0.001).Pearson correlation analysis showed that there was a positive correlation between Nox-4 mRNA expression and TRPV-1,TRPV-4,ASIC-1,ASIC-2,ASIC-3 mRNA expression in st

关 键 词：应激 心理学 酸敏感受体 NADPH氧化酶-4 食管黏膜 炎症 

分 类 号：R571[医药卫生—消化系统]