摘要
背景:1,6-二磷酸果糖是细胞内糖代谢的中间产物,具有改善细胞能量代谢、稳定生物膜、抑制炎症细胞因子释放和抗氧化等多种生物学作用。目的:观察内毒素致大耳白兔急性肺损伤过程中氧化应激反应的变化和1,6-二磷酸果糖的拮抗作用,探讨1,6-二磷酸果糖对急性肺损伤的治疗性作用。设计:完全随机分组设计,对照实验。单位:解放军总医院呼吸科。材料:实验于2003-05/2003-12在解放军总医院呼吸科实验室完成。选用清洁级雄性大耳白兔24只。随机将兔分为3组:对照组、致伤组和干预组,每组8只。方法:①对照组:静脉注射生理盐水2mL/kg。致伤组:将内毒素按500μg/kg溶于2mL生理盐水后于5min内经颈静脉插管一次注射完毕,随后注射生理盐水1次(两次液体总量为2mL/kg)。干预组:内毒素使用剂量和方法同致伤组,随后缓慢经静脉注射1,6-二磷酸果糖(300mg/kg)的生理盐水溶液(2次液体总量为2mL/kg)。②各组均于实验0,0.5,2,4,6h后采外周血和动脉血,采用自动血细胞分析仪进行血细胞计数,采用血气分析仪进行血气分析。测定实验6h末,分别采用巴比妥酸比色法和邻苯三酚自氧化抑制法测定兔肺组织中脂质过氧化物的含量和超氧化物歧化酶的活性。另取右侧膈叶肺组织,采用透射电镜做病理学观察。③符合正态分布和方差齐性等条件者做t检验,否则,做秩和检验。主要观察指标:①实验前及实验0.5,2,4,6h后各组兔血气分析和血细胞计数测定结果比较。②实验后6h兔肺组织中脂质过氧化物含量、超氧化物歧化酶活性和肺组织病理学变化。结果:①实验0.5h后,致伤组动物血氧分压和外周血白细胞计数明显低于对照组(t=-4.27,P<0.01,z=2.64,P<0.01);干预组血氧分压和外周血白细胞计数与对照组差异不明显(P>0.05)鸦干预组血氧分压明显高于致伤组(t=4.32,P<0.01)。②实验4h后,干预组动脉血氧分压明显高于对照组和致伤组(t=4.98,2.40,P<0.01,0.05),致伤组外周血白细胞计数明显低于对照组(z=2.42,P<0.05)。③实验6h后,干预组动脉血氧分压明显高于对照组(t=3.39,P<0.01),致伤组外周血白细胞计数明显低于对照组(z=2.16,P<0.05)。致伤组肺组织脂质过氧化物含量明显高于对照组(t=3.70,P<0.01)、超氧化物歧化酶活性明显低于对照组(t=-4.12,P<0.01),肺组织出现明显的病理损害。实验6h后,干预组肺组织脂质过氧化物含量和超氧化物歧化酶活性与对照组比较,差异不明显(P>0.05),肺组织病理损害较轻。结论:机体氧化应激反应能力的相对不足可以加重氧化损伤,是急性肺损伤发病过程中的重要致病因素;1,6-二磷酸果糖可改善机体的氧化应激反应能力、抑制氧化损伤,对内毒素所致的大耳白兔急性肺损伤具有一定的保护作用。
BACKGROUND： Fructose-1, 6-diphosphate （FDP） is an intermediate product in the course of cellular glycometabolism, and has many biological effects, such as improving cellular energy metabolism, stabilizing biological membrane, inhibiting inflammation intermedium release, antagonizing oxidation, and so on. OBJECTIVE： To observe the changes of oxidative stress reaction in the course of endotoxin （ET） induced acute lung injury （ALI） in flap-eared white rabbits and the antagonistic action of FDP, and to explore the therapeutic effects of FDP on ALI. DESIGN： Complete randomized group design, controlled experiment. SETTING： Department of Respiratory Medicine, General Hospital of Chinese PLA. MATERIALS： The experiment was conducted in the laboratory of Department of Respiratory Medicine, General Hospital of Chinese PLA between May and December 2003, Twenty-four cleaning male flap-eared white rabbits were selected and randomly assigned into control group, injured group and interventional group with 8 animals in each group. METHODS： ①Control group： The rabbits were injected with saline （2 mL/kg） through vein. Injured group： The rabbits were injected with ET （500μg/kg） solution dissolved in 2 mL saline through cannulation in cervical vein once within 5 minutes, and then saline was injected once （Total amount of the solution was 2 mL/kg）. Interventional group： The rabbits were injected with the same amounts of ET solution within 5 minutes as that in the injured group, but FDP （300 mg/kg） solution was injected later （Total amount of solution was also 2 ml/kg）. ②Peripheral blood and arterial blood were collected at 0, 0.5, 2, 4 and 6 hours in each group. Peripheral blood cell count and arterial blood gas were measured with autoblood cell analyzer and blood gas analyzer, respectively. After rabbits were sacrificed through bloodletting at the 6^th hour, content of lipid peroxide （LPO） and activity of superoxide dismutase （SOD） in the pulmonary tissues of rabbits were measured with barbituric acid colorimetry and pyrogallol auto-oxidizing suppression, respectively. In addition, some right lower lobes of pulmonary tissues were observed on pathology with transmission electron microscope （TEM）.③Data accorded with normal distribution and homogeneity of variance were compared by t-test, or by rank sum test. MAIN OUTCOME MEASURES： ①Comparison of result of arterial blood gas and blood cell count measured before experiment, after 0.5, 2, 4 and 6 hours experiment in rabbits of each group. ②Rabbits were sacrificed after 6 hours experiment, the pulmonary contents of LPO and the activities of SOD were measured, and the pathologic changes were also examined. RESULTS： ①At 0.5 hour after ET injected, arterial blood oxygen pressure and white blood cell count in peripheral blood were lower significantly in the rabbits of the injured group as compared with those of the control group （t=-4.27, P 〈 0.01 ,z=2.64, P 〈 0.01 ）. Arterial blood oxygen pres- sure and white blood cell count in peripheral blood in the interventional group showed insignificant difference as compared with the control group （P 〉 0.05 ）. The arterial blood oxygen pressure in the interventional group was obviously higher than that in the injured group （t=4.32,P 〈 0.01 ）.② At the 4^th hour after experiment, the arterial blood oxygen pressure in the interventional group was distinctly higher than those of the control group and the injured group （t=4.98, 2.40, P 〈 0.01, 0.05）. The white blood cell count in the injured group was dramatically less than those of the control group （z=2.42,P 〈 0.05）. ③At the 6^th hours after experiment, the arterial blood oxygen pressure in the interventional group was markedly higher than those of the control group （t=3.39,P 〈 0.01）, The white blood cell count in the injured group was remarkably less than those of the control group （z=2.16,P〈 0.05）. Content of pulmonary LPO in the injured group was significantly higher than that in the control group （t=3.70,P 〈 0.01） while the activity of SOD was obviously lower than that in the control group （t=-4.12,P 〈 0.01）. Markedly pathological lesion appeared in the pulmonary tissue. Six hours later, the content of pulmonary LPO and the activity of pulmonary SOD in the interventional group had no remarkable difference from those in the control group （P 〉 0.05）, and the pulmonary pathologic injury showed less obvious. CONCLUSION： The relative deficiency of oxidative stress reaction can aggravate oxidative injury, which is an important role in pathogenesis of ET-induced ALl. FDP can improve abilities of oxidative stress reaction, inhibit oxidative injury and protect rabbits from ET-induced ALI in flapeared rabbits to a certain extent.
出处
《中国临床康复》
CSCD
北大核心
2006年第12期190-192,F0003,共4页
Chinese Journal of Clinical Rehabilitation
