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目的建立斑马鱼的急性缺氧模型,并对五环三萜类化合物的抗缺氧活性进行初筛。方法 64只斑马鱼放置低氧操作箱中建立斑马鱼的急性缺氧模型,并随机分成空白组、阳性对照组[槲皮素(QT)]、积雪草酸(AA)组、羟基积雪草酸(MA)组、积雪草苷(AS)组、羟基积雪草苷(MS)组、熊果酸(UA)组、甘草次酸(GA)组,每组8只。实验组和阳性对照组所加药物的终浓度均为2.5μg·mL~(-1),空白组为加入相同剂量的20%PEG-400溶液。比较各组斑马鱼的生理状态和半数死亡(4只)所用时间。结果斑马鱼的缺氧条件为20℃,3%的氧气浓度。空白组、阳性对照组、AA组、MA组、AS组、MS组、UA组、GA组的半数死亡所用时间分别为(126.22±4.92),(166.98±3.30),(157.02±4.62),(181.02±3.54),(118.98±3.78),(121.98±1.98),(127.98±2.82),(129.48±5.94)min,除AS组和GA组外,其他组斑马鱼半数死亡时间与空白组比较,差异均有统计学意义(P<0.05)。结论用斑马鱼急性缺氧模型对潜在抗缺氧活性物质的筛选评价建模容易、结果可靠;三萜类苷元(AA、MA、UA和GA)具有抗缺氧作用,且乌苏烷型三萜(AA、MA和UA)的抗缺氧作用更明显;三萜类皂苷(AS、MS)不具有抗缺氧活性。
Objective To establish a model of acute hypoxia in zebrafish and screen the antitumor activity of pentacyclic triterpenoids. Methods A total of 64 zebrafish were placed in hypoxia operation box to establish acute hypoxia model of zebrafish and randomly divided into blank group, positive control group (quercetin (QT), asiatic acid (AA) group, MA group, asiaticoside group, MS group, UA group and GA group. There were 8 rats in each group. The final concentration of drug added in experimental group and positive control group was 2.5μg · mL -1, while in blank group, the same dosage of 20% PEG-400 solution was added. The physiological status of zebrafish in each group and the time spent on half of them (4) were compared. Results The anaerobic condition of zebrafish was 20 ℃, 3% oxygen concentration. The half-time of death in blank group, positive control group, AA group, MA group, AS group, MS group, UA group and GA group were (126.22 ± 4.92), (166.98 ± 3.30), (157.02 ± 4.62) The mean time to death of zebrafish in the other groups was significantly higher than that in the blank group except for AS group and GA group (181.02 ± 3.54), (118.98 ± 3.78), (121.98 ± 1.98), (127.98 ± 2.82) and (129.48 ± 5.94) The differences were statistically significant (P <0.05). Conclusions The zebrafish acute hypoxia model can be used to model potential hypoxia-active substances easily and the results are reliable. Triterpene glycosides (AA, MA, UA and GA) have anti-hypoxia effect, Triterpenoids (AA, MA and UA) were more effective against hypoxia. Triterpenoid saponins (AS, MS) did not have anti-hypoxic activity.