乙酰唑胺对低氧应激下SD大鼠肺泡膜通透性的影响  

Effects of acetazolamide on alveolar membrane permeability in rats under hypoxic stress

作　　者：杜忠蕾 Muhammad Shoaib Tahir 高源 马倩倩 朱沁芳 刘杰[1] 刘美恒 张欢[2] 张伟[1,2] DU Zhonglei;Muhammad Shoaib Tahir;GAO Yuan;MA Qianqian;ZHU Qinfang;LIU Jie;LIU Meiheng;ZHANG Huan;ZHANG Wei(Medical College of Qinghai University,Xining 810001,Qinghai Province,China;Research Center for High Altitude Medicine,Qinghai University,Xining 810001,Qinghai Province,China;Mudanjiang Second People's Hospital,Mudanjiang 157000,Heilongjiang Province,China;Huai'an First People's Hospital,Huai'an 223300,Jiangsu Province,China)

机构地区：[1]青海大学医学院,青海西宁810001 [2]高原医学教育部重点实验室,青海西宁810001 [3]黑龙江省牡丹江市第二人民医院,黑龙江牡丹江157000 [4]江苏省淮安市第一人民医院,江苏淮安223300

出　　处：《解放军医学院学报》2022年第3期340-346,共7页Academic Journal of Chinese PLA Medical School

基　　金：国家自然科学基金项目(81560301);青海省科技厅项目(2022-ZJ-905)

摘　　要：背景急进高原容易产生高原肺水肿(high altitude pulmonary edema,HAPE)。乙酰唑胺作为国际通用预防急性高原病的首选药,在低氧应激过程中的调节作用尚不清楚,尤其对肺泡膜通透性的影响还不明确。目的研究乙酰唑胺对低氧应激下SD大鼠肺泡膜通透性的影响。方法将雄性SD大鼠随机分为4组(每组12只):对照组(CN组,西宁海拔2260 m)、低氧组(H7K组,模拟海拔7000 m)、乙酰唑胺组(NAZ组,西宁海拔2260 m)、低氧+乙酰唑胺组(HAZ组,模拟海拔7000 m)。NAZ、HAZ组给予1%乙酰唑胺(50 mg/kg),CN、H7K组给予等容量的0.9%氯化钠注射液(45 mg/kg),灌胃(Bid,共5 d)。灌胃3 d后将H7K、HAZ组大鼠置于低压氧舱中2 d(模拟海拔7000 m,大气压305 mmHg,PO_(2)63.7 mmHg;1 mmHg=0.133 kPa)。测定大鼠动脉pH、二氧化碳分压(PaCO_(2))、氧分压(PaO_(2))、碳酸氢盐(HCO_(3)^(-))等;测定右肺组织湿重/干重(W/D);ELISA测定支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)及血清白细胞介素6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)含量;qRT-PCR检测左肺组织水通道蛋白1(aquaporin 1,AQP1)、AQP5的mRNA表达量;免疫组化检测肺组织中AQP1和AQP5的表达水平。观察各组肺组织形态结构变化。结果与CN组比较,NAZ组pH、PaCO_(2)下降;H7K组动脉血pH值下降,PaO_(2)降低(P<0.05),右肺组织的W/D升高;BALF及血清中IL-6、TNF-α含量增高;左肺组织的AQP1、AQP5 mRNA及蛋白表达量均升高(P<0.05)。H7K组的肺组织切片HE染色见肺泡上皮细胞肿胀明显,肺泡间隔增宽伴渗出。与H7K组比较,HAZ组动脉血PaO_(2)升高(P<0.05);W/D比值降低(P<0.05);BALF中IL-6含量降低,血清中IL-6、TNF-α含量降低(P<0.05)。左肺组织的AQP1、AQP5 mRNA及蛋白表达量减少(P<0.05)。HAZ组的肺组织切片HE染色可见肺泡上皮细胞偶有肿胀,肺泡间隔略增宽,肺水肿程度减轻。结论乙酰唑胺预处理能减少IL-6、TNF-α的释放和AQP1、AQP5的Background Acute exposed to high altitude areas can result in high altitude pulmonary edema(HAPE).Acetazolamide,as the internationally accepted drug of the first choice for the prevention of acute mountain sickness,has unclear regulatory effects in the process of hypoxic stress,especially its effect on alveolar membrane permeability.Objective To observe the effect of acetazolamide on alveolar capillary membrane permeability in SD rats with acute hypoxia.Methods Male SD rats were randomly divided into control group(CN group,Xining altitude 2260 m,n=12),hypoxia 7 km group(H7K group,simulated altitude of 7000 m,n=12),acetazolamide group(NAZ group,Xining altitude 2260 m,n=12),hypoxia and acetazolamide group(HAZ group,simulated altitude of 7000 m,n=12).NAZ group and HAZ group were given 1%acetazolamide(50 mg/kg).CN and H7K groups were given equal volume of 0.9%sodium chloride injection(45 mg/kg)twice a day for 5 days.The H7K and HAZ groups were placed in a low pressure simulation chamber in an altitude of 7000 m(Barometric pressure 305mmHg,PO_(2)63.7 mmHg)for 2 days.Arterial blood gas analysis was performed to measure arterial blood[pH,partial pressure of carbon dioxide(PaCO_(2)),partial pressure of oxygen(PaO_(2))and bicarbonate(HCO_(3)^(-))].The wet/dry weight ratio(W/D)of right lung tissue was determined.The contents of interleukin(IL-6)and tumor necrosis factor(TNF-α)in bronchoalveolar lavage fluid(BALF)and serum were tested.The mRNA expression levels of aquaporin 1(AQP1)and aquaporin 5(AQP5)in lung tissue were detected by qRT-PCR.The morphological and structural changes of lung tissues were compared,and the expression levels of AQP1 and AQP5 in lung tissues of rats in each group were detected by immunohistochemistry.Results Compared with the CN group,the arterial blood pH and PaO_(2) in NAZ group and H7K group decreased significantly(P<0.05),while the W/D increased significantly(P<0.05).The contents of IL-6 and TNF-αin BALF and serum increased,and AQP1,AQP5 mRNA and protein expressions in the right lung tissue

关 键 词：低氧 应激 肺泡膜 通透性 乙酰唑胺 水通道蛋白 

分 类 号：R743.3[医药卫生—神经病学与精神病学]