玉竹提取物A对内毒素血症小鼠血清中肿瘤坏死因子α及一氧化氮水平影响的量效依赖性

Dose-dependent effect of fragrant solomonseal rhizome extract A on serum tumor necrosis factor alpha and nitrogen monoxide of mice with endotoxemia

目的:观察百合科黄精属中草药玉竹的醇提取物玉竹提取物A对内毒素血症小鼠72h存活率、血清中肿瘤坏死因子及一氧化氮水平的影响。方法:①实验于2005-01/03在锦州医学院免疫学实验室完成。选用6～8周龄雄性昆明种小鼠150只。150只小鼠被随机分为2组:Ⅰ组100只,Ⅱ组50只。②将Ⅰ组小鼠随机分为4组:模型组和玉竹提取物A(由锦州医学院潘兴瑜教授提供)低、中、高剂量组,每组25只。玉竹提取物A低、中、高剂量组腹腔注射玉竹提取物A0.5,1,2g/kg,1次/d,连续7d,模型组腹腔注射等量生理盐水。末次给药后1h模型组和玉竹提取物A组应用内毒素腹腔注射(10mg/kg)建立小鼠内毒素血症模型,记录小鼠72h存活率。③Ⅱ组小鼠随机分为5组:正常对照组,模型组和玉竹提取物A低、中、高剂量组,每组10只。模型组和玉竹提取物A低、中、高剂量组干预措施同Ⅰ组,正常对照组给予等量生理盐水。内毒素给药后6h采血并应用双抗体夹心酶联免疫吸附实验检测各组小鼠血清肿瘤坏死因子α水平,化学比色硝酸还原酶法检测血清一氧化氮水平。④组间显著性检验应用单向方差分析,组间比较方差齐时选用LSD法,方差不齐时用DunnetT3法。结果:昆明种小鼠150只均进入结果分析。①玉竹提取物A各剂量组72h生存率均明显高于模型组(P<0.05),玉竹提取物A各剂量组间72h生存率比较,差异不明显(P>0.05)。②模型组血清肿瘤坏死因子α和一氧化氮水平明显高于正常对照组(P<0.01)。玉竹提取物A低、中、高剂量组血清肿瘤坏死因子α和一氧化氮水平明显低于模型组,且呈量效依赖性(P<0.01),玉竹提取物A中、高剂量组血清一氧化氮水平明显低于模型组(P<0.01),但玉竹提取物A低剂量组血清一氧化氮水平与模型组比较,差异不明显(P>0.05)。结论:玉竹提取物A能够显著提高内毒素血症小鼠72h存活率;玉竹提取物A可明显降低血清中肿瘤坏死因子α和一氧化氮水平,且呈量效依赖性。

AIM： To observe the effect of liliaceous herb ethanol extraction, fragrant solomonseal rhizome extract A （EA-PAOA）, on the 72 hour survival rate, serum tumor neerasis factor alpha （TNF-α） and nitrogen monoxide （NO） levels of mice with endotoxemia. METHODS： The experiment was completed in the Laboratory of Immunology, Jinzhou Medical College from January to March 2005. （1） Totally 150 male kunming mice aged 6-8 weeks were divided into group I with 100 mice and group Ⅱ with 50 mice.（2） Mice in group I were divided into 4 groups： model group and EA-PAOA （provided by Pro. Pan Xing-yu from Jinzhou Medical College） groups of low, moderate and high dosage with 25 in each group. Mice in EA-PAOA groups were intravenously injected with 0.5, 1 and 2 g./kg EA-PAOA once a day for 7 successive days respectively, and mice in model group were intravenously injected with the equal volume of saline. One hour after the last medication, mice in model group and EA-PAOA group were intravenously injected with 10 mg/kg endotoxin to establish endotoxemia model so as to record 72-hour survival rate of mice. （3） Mice in group 11 were divided randomly into 5 groups： normal control group, model group and EA-PAOA groups of low, moderate and high dosages with 10 in each group. Administration in model group and EA-PAOA groups were the same as that in group I, and mice in normal control group were treated with the equal volume of saline. Levels of TNF-α in serum were quantified by sandwich enzyme-linked immunoadsordent assay （ELISA） method, and NO level was detected by nitrate reductase chemical colorimetry 6 hours after injection of endotoxin. （4） One way analysis of variance was employed in significance test of different groups, followed by LSD analysis with equal variances and Dunnet T3 test with unequal variances. RESULTS：All 150 kunming mice entered the final analysis. （1） 72-hour survival rates in EA-PAOA grouPs were higher than those in model group （P 〈 0.05）, and no significant difference at three EA-PAOA dose groups was shown （P 〉 0.05）. （2） In model group, levels of TNF-ct and NO were higher than those in normal control group （P 〈 0.01）, but those in EA- PAOA dosage groups were lower than those in model group which showed dosage dependence （P 〈 0.01）. NO levels in moderate and high EA-PAOA groups were lower than those in model group （P 〈 0~01）, but difference of NO level in low EA-PAOA group was not significant （P 〉 0.05）. CONCLUSION： EA-PAOA can increase 72-hour survival rate of mice with endotoxemia, and decrease levels of TNF-ct and NO, which shows dose dependence.