天麻种子抗疲劳和耐缺氧作用及其脂溶性成分研究

Anti-fatigue and anti-hypoxia ability and liposoluble constituents of Gastrodia elata seeds

【目的】对天麻种子脂溶性成分进行分析并研究天麻种子的抗疲劳和耐缺氧作用,为其开发利用提供依据。【方法】采用索氏提取法提取天麻种子的脂溶性成分,用GC-MS对天麻种子脂溶性成分进行分析。将100只昆明种小鼠,随机分为阴性对照组(每10g体质量灌胃5g/L羧甲基纤维素钠溶液0.2mL)、刺五加颗粒组(灌胃刺五加颗粒0.75g/kg)及天麻种子高剂量组(灌胃天麻种子4g/kg)、中剂量组(灌胃天麻种子2g/kg)、低剂量组(灌胃天麻种子1g/kg),每天08:00灌胃给药1次,连续给药21d。通过负重游泳试验测定小鼠力竭游泳时间,并检测小鼠血清乳酸(BLA)、尿素氮(BUN)、肝糖原浓度和乳酸脱氢酶(LDH)活性。另将120只小鼠随机分为阴性对照组和天麻种子高、中、低剂量组,相同剂量连续灌胃给药28d后,测定小鼠常压耐缺氧存活时间及血清中超氧化物歧化酶(SOD)质量浓度、丙二醛(MDA)浓度、亚硝酸钠中毒存活时间和急性脑缺血性缺氧存活时间。【结果】采用GC-MS对天麻种子脂溶性成分进行分析,共鉴定出醇、醛、酯、烷烃和脂肪酸等5类25种物质,其中2-甲基十八烷、3-乙基-5-(2-乙基丁基)-十八烷、2-甲基-1-十一烷醇、己醛、己酸和鱼鲨烯相对含量较高,分别为20.79%,15.10%,14.77%,11.69%,7.96%和4.79%。抗疲劳试验表明,天麻种子可以极显著延长小鼠力竭游泳时间(P<0.01);与阴性对照组相比,天麻种子高、中剂量组小鼠血清中的BLA、BUN浓度显著降低(P<0.05),高剂量组小鼠血清中的LDH活性和肝糖原浓度极显著升高(P<0.01)。耐缺氧试验显示,与阴性对照组相比,天麻种子高、中剂量组小鼠亚硝酸钠中毒和急性脑缺血性缺氧存活时间显著延长,MDA浓度显著降低(P<0.05),小鼠常压耐缺氧存活时间极显著延长(P<0.01),且天麻种子高剂量组小鼠血清中的SOD质量浓度显著提高(P<0.05)。【结论】天麻种子具有抗疲劳和耐缺氧作用。

[Objective] This study analysis the liposoluble constituents of Gastrodia clara seeds and in- vestigated the anti-fatigue and anti-hypoxia effect in mice to provide the basis for its development and utili- zation. [Method] The liposoluble constituents of Gastrodia clara seeds were extracted by soxhlet extrac- tion,and their constituents were determined by GC-MS. A total of 100 mice were randomly divided into negative control group （5 g/L carboxymethyl cellulose sodium solution was 0.2 mL per 10 body mass） ,ac-anthopanax particles group （Acanthopanax particles,0. 75 g/kg）,Gastrodia elata seeds high dose group （Gastrodia elata seeds,4 g/kg）, medium dose group （Gastrodia elata seeds, 2 g/kg） and low dose group （Gastrodia elata seeds,1 g/kg）. Drug was administrated to mice once every day for 21 days. The exhaus- tive swimming time and the concentration of BLA,BUN,hepatic glycogen and the activity of LDH of mice were determined by weight loading swimming experiment. Another 120 mice were randomly divided into negative control group,and Gastrodia elata seeds high,medium and low dose groups,and same drug was administrated to mice once every day for 28 days. The hypoxia tolerant survival time of mice and the con- centration of SOD and MDA, sodium nitrite poisoning survival time and acute cerebral hypoxia survival time were measured. ~Resultl A total of 25 compounds liposoluble constituents of Gastrodia elata seeds were found by GC-MS including alcohols, aldehydes, esters, alkane and fatty acid. The 2-methyl-octade- cane, 3-ethyl-5-（2-ethylbutyl）-octadecane, 2-methyl-l-undecanol, hexanal, hexanoic acid and squalene had high relative contents of 20.79% ,15.10% ,14.77% ,11.69% ,7.96% and 4.79%. Anti-fatigue experiment showed that Gastrodia elata seeds could prolong the exhaustive swimming time significantly （P〈0.01）. The high dose group and medium dose group reduced the concentrations of serum BLA and BUN markedly （P〈0.05）, and the high dose group increased LDH activity and hepatic glycogen concentration significant- ly （P〈0.01）. Anti-hypoxia experiment showed that high and medium dose groups could markedly prolong the sodium nitrite poisoning survival time and survival time in acute cerebral hypoxia, markedly reduce MDA concentration （P〈 0. 05）, and prolong the survival time in hypoxia tolerance significantly （P 0.01）. The high dose group could also increase SOD concentration markedly （P〈0.05）.[ Conclusion] The Gastrodia elata seeds has anti-fatigue and anti-hypoxia function.