CAJ | 학술논문

目的评价Toll样受体4(TLR4)在机械通气致糖尿病小鼠肺毛细血管通透性增加中的作用.方法选择雄性野生型小鼠C3H/HeN和TLR4基因突变型小鼠C3H/HeJ各28只,8～ 10周龄,体重20 ～ 25 g,腹腔注射链脲佐菌素150 mg/kg的方法制备糖尿病模型.采用随机数字表法,将2种小鼠各随机分为2组(n=14),假手术组(DS组)糖尿病小鼠行气管插管术自主呼吸4h;机械通气组(DM组)糖尿病小鼠进行机械通气4h,吸入氧浓度50％,通气频率70次/min,潮气量15 ml/kg,PEEP 2cmH2O.自主呼吸或机械通气4h时采集颈动脉血样行血气分析,记录PaO2和PaCO2,收集血清,测定血清一氧化氮(NO)浓度.随后处死,取肺组织测定肺组织湿/干重比(W/D比值)、伊文思蓝(EB)含量,Western blot法测定小凹蛋白-1、内皮型一氧化氮合成酶(eNOS)和磷酸化内皮型一氧化氮合成酶(p-eNOS)的蛋白表达水平,计算p-eNOS/eNOS比值.结果与DS组比较,DM组小鼠PaO2、PaCO2、p-eNOS/eNOS比值降低,肺组织W/D比值、EB含量、小凹蛋白-1表达水平明显升高(P＜0.05);与C3H/HeN小鼠比较,DM组C3H/HeJ小鼠肺组织W/D比值、EB含量和小凹蛋白-1表达水平明显降低,p-eNOS/eNOS比值和血清NO浓度升高(P＜0.05),PaO2和PaCO2差异无统计学意义(P＞0.05).结论 TLR4通过与小凹蛋白-1有效结合,减少NO生物利用度,增加糖尿病小鼠机械通气后肺毛细血管通透性.
목적 평개Toll양수체4(TLR4)재궤계통기치당뇨병소서폐모세혈관통투성증가중적작용.방법 선택웅성야생형소서C3H/HeN화TLR4기인돌변형소서C3H/HeJ각28지,8～ 10주령,체중20 ～ 25 g,복강주사련뇨좌균소150 mg/kg적방법제비당뇨병모형.채용수궤수자표법,장2충소서각수궤분위2조(n=14),가수술조(DS조)당뇨병소서행기관삽관술자주호흡4h;궤계통기조(DM조)당뇨병소서진행궤계통기4h,흡입양농도50％,통기빈솔70차/min,조기량15 ml/kg,PEEP 2cmH2O.자주호흡혹궤계통기4h시채집경동맥혈양행혈기분석,기록PaO2화PaCO2,수집혈청,측정혈청일양화담(NO)농도.수후처사,취폐조직측정폐조직습/간중비(W/D비치)、이문사람(EB)함량,Western blot법측정소요단백-1、내피형일양화담합성매(eNOS)화린산화내피형일양화담합성매(p-eNOS)적단백표체수평,계산p-eNOS/eNOS비치.결과 여DS조비교,DM조소서PaO2、PaCO2、p-eNOS/eNOS비치강저,폐조직W/D비치、EB함량、소요단백-1표체수평명현승고(P＜0.05);여C3H/HeN소서비교,DM조C3H/HeJ소서폐조직W/D비치、EB함량화소요단백-1표체수평명현강저,p-eNOS/eNOS비치화혈청NO농도승고(P＜0.05),PaO2화PaCO2차이무통계학의의(P＞0.05).결론 TLR4통과여소요단백-1유효결합,감소NO생물이용도,증가당뇨병소서궤계통기후폐모세혈관통투성.
Objective To evaluate the role of Toll-like receptor 4 (TLR4) in mechanical ventilationinduced increase in pulmonary capillary permeability in diabetic mice.Methods Two types of mice,aged 8-10 weeks,weighing 20-25 g,were used in this study:free wild type mice (C3H/HeN) and TLR4 gene mutation type (C3H/HeJ).Diabetes was induced by intraperitoneal streptozotocin 150 mg/kg (in citric acid buffer solution 0.1 mol/L) and confirmed by blood glucose level ＞ 16 mmol/L.Each type of diabetic mice was randomly divided into 2 groups (n =14 each):group diabetes + sham operation (group DS) and group diabetes + mechanical ventilation (group DM).The animals were anesthetized with intraperitoneal ketamine,midazolam and atropine.Tracheal intubation was performed and the animals kept spontaneous breathing in group DS.The animals were mechanically ventilated (FiO2 50％,RR 70 bpm,VT 15 ml/kg,PEEP 2 cmH2O) for 4 h in group DM.Arterial blood samples were obtained at 4 h of ventilation for blood gas analysis,PaO2 and PaCO2 were recorded,and NO concentration in the serum was measured.Then the animals were sacrificed and the lungs were removed for determination of W/D lung weight ratio (W/D ratio),content of evans blue (EB),and expression of caveolin-1,endothelial nitric oxide synthase (eNOS) and phosphor-eNOS (p-eNOS) (by Western blot).The ratio of p-eNOS/eNOS was calculated.Results Compared with group DS,PaO2,PaCO2 and p-eNOS/eNOS ratio were significantly decreased,and W/D ratio,EB content and caveolin-1 expression were increased in group DM (P ＜ 0.05).Compared with C3H/HeN mice,W/D ratio,caveolin-1 expression and EB content were significantly decreased,peNOS/eNOS ratio and serum NO concentrations were increased (P ＜ 0.05),and no significant changes in PaO2 and PaCO2 were found in C3H/HeJ mice in group DM (P ＞ 0.05).Conclusion TLR4 increases the pulmonary capillary permeability after mechanical ventilation in diabetic mice through effectively combining with caveolin-1 and reducing NO bioavailability.