冬虫夏草氨基酸成分的药理作用分析

Analysis of amino acid composition and the pharmacology of cordyceps sinensis

目的分析冬虫夏草氨基酸成分药理作用。方法取中药研究院人工培育的冬虫夏草,常规预处理,取药粉。健康雄性清洁小鼠,7 w龄,进行训练,取出与均值差异较大个体。取药粉,烘干,烘烤水解,预处理后以全自动氨基酸分析仪检查,天冬氨酸、丝氨酸等含量在0.5%~3.5%之间。取运动能力无明显差异的小鼠100只,随机分为5组,分别为模型组,空白对照组,高、中、低剂量氨基组。除模型组外,其余均灌胃给药4 w,每次0.4 ml;空白对照组为生理盐水;模型组仅进行训练,不进行灌胃。高、中、低氨基组分别给予高、中、低剂量氨基酸0.4、0.2、0.1 ml;氨基酸参照前文提到的氨基酸配比,混悬在羧甲基纤维素钠中。4 w后,末次给药前进行游泳力竭训练,继续灌胃2 d,末次灌胃30 min,进行游泳40 min,休息15 min,摘眼球采血0.1 ml,进行全血乳酸含量检测。同法分组,除空白对照组外,其余腹腔注射环磷酰胺50 mg/kg,制作小鼠免疫低下模型; 1次/d,连续2 d,灌胃2 w后,末次给药20 min,称重,摘除眼球取血,脱颈法处死;取出脾脏、胸腺称重,计算脏器指数,检测小鼠白细胞介素(IL-2)、肿瘤坏死因子α(TNF-α)水平。结果氨基酸组可明显增强小鼠的抗疲劳能力,力竭游泳时间从0.7 min增加到10.4 min。小鼠游泳时间明显延长,同时血清乳酸水平下降,从5.1 mmol/L下降到1.3 mmol/L,与此同时脾脏指数、胸腺指数、IL-2、TNF-α水平均有所改善。5组小鼠力竭游泳时间、血乳酸水平、脾脏指数、胸腺指数、IL-2、TNF-α水平比较,差异有统计学意义(均P<0.05),高、中、低氨基酸组指标呈梯度变化,逐渐接近空白对照组,差异有统计学意义(均P<0.05)。结论冬虫夏草氨基酸具有抗疲劳、增强免疫作用。

Objective To analyze the amino acid composition of Nordyceps pharmacology. Methods Cordyceps sinensis,which bred by the traditional Chinese medicine research institute,was routinely pretreated and powder extracted. The healthy male clean mice were trained at 7 weeks old,and the individuals with large differences were taken out. The powder was dried,baked,hydrolyzed and pretreated,followed by automatic amino acid analyzer. The contents of aspartic acid and serine were between 0.5% and 3.5%. 100 mice with no obvious difference in motor ability were randomly divided into 5 groups： model group,blank control group,high,middle and low dose amino group,except the model group,all of them were given intragastric administration for 4 weeks. Each time 0.4 ml,normal saline,model group only for training,not for oral administration,high,medium and low amino groups,respectively,given high,medium and low doses of amino acid0.4 ml,0.2 ml,0.1 ml. According to the proportion of amino acids mentioned above,after suspension in sodium carboxymethyl cellulose for 4 weeks,swimming exhaustion training was performed before the last administration,and continued to be administered by stomach for 30 minutes on 2 nd and at the last time. After swimming for 40 minutes,rest for 15 minutes,blood was taken from eyeball 0.1 ml,the content of lactic acid in whole blood was detected. In the same method,in addition to the blank control group,the other mice were injected cyclophosphamide 50 mg/kg intraperitoneally to make the model of immunosuppression in mice. 1 st once,two days in a row,after 2 weeks of gavage,the mice were given the last dose of cyclophosphamide for 20 minutes. Weighing,removing eyeball and blood,removing neck,removing spleen,weighing thymus,calculating viscera index,examining interleukin-2（ IL-2）,tumor necrosis factor α（ TNF-α）. Results In the amino acid group,the anti-fatigue ability of mice was significantly enhanced. the exhaustive swimming time increased from 0.7 min to 10.4 min. the swimming time of mice was significantly prolonged,while the serum lactic acid level decreased from 5.1 mmol/L to 1.3 mmol/L. meanwhile,the spleen index,thymus index,il-2,TNF-α levels were improved.The levels of lactic acid,spleen index,thymus index,interleukin-2（ IL-2） and interleukin-α（ TNF-α） in five groups of mice were statistically significant（ P〈0.05）,and the difference was statistically significant（ P 0.05）.Conclusion Cordyceps sinensis amino acids have anti-fatigue and immunity enhancement.