佛甲草抗疲劳作用的动物实验

An animal experiment on fatigue-resisting action of sedum lineare

目的观察佛甲草提取液对小鼠综合体能及其小鼠血清、肝组织丙二醛、超氧化物歧化酶的影响,从而探讨该提取液的抗疲劳作用。方法①实验于2005-04/05在赣南医学院机能实验室完成。选用昆明种小鼠100只,雌雄不拘,体质量(20±2)g。②观察佛甲草提取液(由佛甲草提取所得)对小鼠耐寒能力的影响取雄性小鼠20只,随机分成2组空白对照组和佛甲草实验组,每组10只。空白对照组腹腔注射生理盐水10m L/kg;佛甲草组腹腔注射佛甲草提取液5g/kg。给药后1h,将小鼠置于-20°C低温冰箱中,记录小鼠存活时间。③观察佛甲草提取液对小鼠耐热能力的影响取雌性小鼠20只,随机分为2组,每组10只。空白对照组腹腔注射空白对照10m L/kg;佛甲草组腹腔注射佛甲草提取液5g/kg。给药后1h,将小鼠置于50℃恒温箱中,记录小鼠存活时间。④观察佛甲草提取液对小鼠负重游泳时间的影响取雄性小鼠20只,随机分为2组空白对照组和佛甲草组,每组10只。空白对照组腹腔注射空白对照10m L/kg;佛甲草组腹腔注射佛甲草提取液5g/kg。给药后1h,小鼠按体质量的5%尾根铜丝负重,置25℃恒温水箱中,记录小鼠从入水至沉入水中不再浮出水面为止所需时间,即为小鼠游泳时间。⑤观察佛甲草提取液对小鼠血清、肝组织丙二醛含量和超氧化物歧化酶活性的影响将其余40只小鼠随机分为4组空白对照对照组、维生素E对照组,佛甲草低剂量组和佛甲草高剂量组,每组10只。空白对照组小鼠灌胃空白对照10m L/(kg·d);维生素E对照组灌胃维生素E2.5g/(kg·d);佛甲草低剂量组灌胃佛甲草提取液5g/(kg·d);佛甲草高剂量组灌胃佛甲草提取液10g/(kg·d),连续14d。末次给药后2h,将各组小鼠断头取血,1500r/m in,离心5m in,取血清;取适量肝组织,用冰冷空白对照在冰浴中制成10%组织匀浆,4℃,3500r/m in,离心10min,取上清液。均采用硫代巴比妥酸比色法和黄嘌呤氧化酶法分别测定血清、肝组织丙二醛含量和超氧化物歧化酶的活性。⑥组间计量资料差异比较采用双侧t检验。结果小鼠100只均进入结果分析。①耐寒存活时间佛甲草组明显长于空白对照组[(58.30±6.88),(48.40±7.93)m in,t=2.9819,P<0.01]。②耐热存活时间佛甲草组明显长于空白对照组[(35.60±6.38),(22.80±6.30)m in,t=4.507,P<0.01]。③负重游泳时间佛甲草组明显长于空白对照组[(74.10±15.47),(43.60±13.62)m in,t=4.6779,P<0.01]。④血清、肝组织丙二醛含量佛甲草提取液低剂量和高剂量组明显低于空白对照组[血清(8.41±1.45),(7.65±1.32),(10.05±1.59)μm ol/L;肝组织(334.12±58.48),(315.13±55.65),(407.82±60.29)nm ol/g,t=2.41,3.6923,2.7748,3.5719,P<0.05～0.01];与维生素E的作用效果基本一致。⑤血清、肝组织超氧化物歧化酶活性佛甲草提取液低剂量和高剂量组明显高于空白对照组[血清(287.63±23.33),(300.86±26.35),(262.52±29.47)kNU/L;肝组织(783.59±27.07),(810.30±38.72),(741.04±45.92)kNU/g,t=2.112,3.0672,2.5237,3.6472,P<0.05～0.01];与维生素E的作用效果基本一致。结论佛甲草提取液具有增强机体活力和适应能力及对抗机体疲劳的作用。

AIM： To observe the effects of sedum lineare extract on the comprehensive physical fitness, the malondialdehyde （MDA） and superoxide dismutase （SOD） in serum and liver of mice, so as to probe into the fatigue-resisting role of sedum lineare extract. METHODS： （1） The experiment was performed in the Functional Laboratory of Gannan Medical College in April and May 2005. A total of 100 Kunming mice （either male or female） weighing （20±2） g were provided by the Experiment Animal Center of Gannan Medical College, the certificated number was JXA-2005019. （2） Observation on the effect of sedum lineare extract on the cold-resistant ability of mice： Twenty male mice were randomly divided into blank control group （n=10） . and sedum lineare extract group （n=10）. Normal saline （10 mL/kg） and sedum lineare extract （5 g/kg） were intraperitoneally injected in the blank control group and sedum lineare extract group respectively. After treatment for 1 hour, each mouse was housed into -20 ℃ refrigerator, and then the survival times were recorded. （3）Observation on the effect of sedum lineare extract on the hot-resistant ability of mice： Twenty female mice were randomly divided into blank control group （n=10） and sedum lineare extract group （n=10）. Normal saline （10 mg/kg） and sedum lineare extract （5 g/kg） were intraperitoneally injected in the two groups respectively. After 1 hour, each mouse was housed into 50 ℃ constant temperature oven, and then the survival times were recorded. （4） Observation on the effect of sedum lineare extract on the loaded swimming time of mice： Twenty male mice were randomly divided into blank control group （n=10） and sedum lineare extract group （n=10）. Normal saline （10 mL/kg） and sedum lineare extract （5 g/kg） were intraperitoneaUy injected in the two groups respectively. After 1 hour, each mouse was leaded with copper wire （5% body mass） at tail, and then placed into 25 ℃ thermostatic water tank, and then the swimming time （from entering water to sinking into water and no longer floating on the water surface） was recorded. （5） Observation on the effect of sedum lineare extract on the MDA content and SOD activity in serum and liver of mice： The rest 40 mice were randomly divided into 4 groups with 10 mice in each group： blank control group, vitamin E control group, sedum. lineare extract high and low dosage groups, and the rats were treated by gastric perfusions of normal saline 10 mL/kg, vitamin E 2.5 g/kg, sedum lineare extract 5 and 10 g/kg per day respectively. At 2 hours after the last administration, the heads of all the mice were cut down to take blood, and then the blood was centrifuged （1 500 r per minute, 5 minutes） to take serum. Certain liver tissue was taken to make 10% tissue homogena± in ice bath with cold saline, and then centrifuged （3 500 r per minute, 10 minutes, at 4 ℃） to take the supernatant. The MDA content and SOD activity were measured with the techniques of thibabituric acid （TBA） fluorescence and xanthine oxidase respectively in serum and liver tissue. （6） The intergroup difference of measurement .data were compared with bilateral t test. RESULTS： All the 100 mice were brought into the analysis of results. （1） The survival time in the cold tolerance test： It was obviously longer in the sedum lineare extract group than in the blank control group [（58.30±6.88）, （48.40±7.93） minutes, t=2.981 9, P 〈 0.01]. （2） The survival time in the hot tolerance test： It ＂was obviously longer in the sedum lineare extract group than in the blank control group [（35.60±6.38）, （22.80±6.30） minutes, t=4.507, P 〈 0.01]. （3） loaded swimming time： It was obviously longer in the sedum lineare extract group than in the blank control group [（74.10 ±15.47）, （43.60±13.62） minutes, t=4.677 9, P 〈 0.01]. （4） MDA content in serum and＇liver tissue： It was obviously lower in the sedum lineare extract low and high dosage groups than in the blank control group [serum： （8.41 ±1.45）, （7.65±1.32）, （10.05±1.59） p, moL/L; （334.12±58.48）, （315.13±55.65）, （407.82±60.29） nmol/g, t=2.41, 3.692 3, 2.774 8, 3.571 9, P 〈 0.05-0.01], which were similar to the results in the vitamin E control group. （5） SOD activity in serum and liver tissue： It was obviously higher in the sedum lineare extract low and high dosage groups than in the blank control group [serum： （287.63±23.33）, （300.86±26.35）, （262.52±29.47） kNU/L; liver tissue： （783.59±27.07）, （810.30±38.72）, （741.04±45.92） kNU/g, t=2.112, 3.067 2, 2.523 7, 3.647 2, P 〈 0.05-0.01], which were similar to the results in the vitamin E control group. CONCLUSION： Sedum lineare extract can enhance the activity and adaptive ability of organ, it also has the effects of anti-fatigue.