摘要
目的：研究小分子多肽化合物CMS 008和CMS 001抗小鼠运动性疲劳的作用及并初步探讨其作用机理；观察小分子多肽化合物CMS 001对慢性疲劳综合症的治疗作用。
方法：1．选用雄性Balb／c小鼠，根据抗疲劳作用的检验方法，观察CMS008，CMS001对小鼠负重游泳时间，肝糖原，尿素氮，血乳酸水平的影响。2．电镜观察CMS 008对运动后小鼠心肌，骨骼肌超微结构的影响，探讨药物对心肌，骨骼肌线粒体，肌浆网的影响。3．观察CMS 008、CMS 001对力竭运动小鼠血清MDA、SOD及AST、ALT水平的影响。4．观察CMS 008、CMS 001对体外淋巴细胞转化的影响。5．通过红外光传感器计数与单片机集中管理数据相结合，设计出24小时记录小鼠日常活动的仪器——动物旋转轮式运动计量系统。6．雌性C57BL／6小鼠，腹腔注射短棒杆菌0.1ml(6.0×10~9个菌／ml)，诱发CFS模型。动物旋转轮式运动计量系统记录CMS 001对CFS小鼠日常活动量的影响。
结果：1．CMS 008给药剂量为125-500μg／kg／d，CMS 001给药剂量为5-20μg／kg／d时，可以明显延长小鼠负重游泳时间(p＜0.05)。2．CMS008给药剂量为125-500μg／kg／d，CMS 001给药剂量为5-20μg／kg／d时，可以明显降低小鼠运动后血清尿素氮水平(p＜0.05)。3．CMS 008给药剂量为125-500μg／kg／d，CMS 001给药剂量为20μg／kg／d时，可以明显增加小鼠肝糖原的储备(p＜0.05)。4．CMS 008给药剂量为125-500μg／kg／d、CMS 001给药剂量为5-20μg／kg／d时，能够抑制运动后乳酸升高并加速机体对乳酸的消除(p＜0.05)。5．CMS 008可以明显减轻运动造成的小鼠心肌，骨骼肌线粒体明显减少，残存线粒体空泡样变，髓磷体沉积及“钙超载”等损伤，使大部分组织接近正常。6．CMS 008给药剂量为500μg／kg／d，CMS 001给药剂量为20μg／kg／d时，可以降低力竭运动时MDA的升高(p＜0.05)，并且能够增加抗氧化酶SOD的活性(p＜0.05)。CMS 008给药剂量为500μg／kg／d，CMS 001给药剂量为20μg／kg／d时，可以
沪丈藏要
降低力竭运动时AST和ALT的升高(p<0.05)。7.CMS 001在2林g/ml，
0.4林创ml，0.08林留ml，0.016林创ml浓度时能够抑制体外淋巴细胞转化
(P<0.ool)，相同浓度的CMS 008对体外淋巴细胞的转化无明显影响
(p>.05)。8.动物旋转轮式运动计量系统可有效记录小鼠的日常活动量。9.
腹腔注射CP后，模型组小鼠在以后的19天内日常活动量下降至原来的50%
以下。CMS 001组与生理盐水组在注射CP后，活动量下降，两组日常活动量
均降到原来的加%以下，两者比较无显著性差异(P>0.05)。
结论:1.cMs 008，cMs 001具有抗小鼠运动性疲劳的作用。2.抗疲劳
作用是通过增加肝糖原储备，抑制运动引起乳酸的堆积，加速乳酸的清除，抑
制蛋白质的分解实现的。另外，药物对心肌、骨骼肌线粒体的保护作用以及减
少线粒体钙超载促进了其抗疲劳作用。药物亦通过减轻运动引起的氧化损伤、
增强机体抗氧化系统酶的活性发挥抗疲劳作用。3.动物旋转轮式运动计量系
统能够记录小鼠24小时日常活动量，可靠、灵敏、简便。CMS 001对CFS小
鼠无治疗作用。
OBJECTION To investigate the anti-fatigue effects and the mechanism of
small molecular peptides CMS 008 and CMS 001 on Balb/c mice, and the effect on CFS of small peptide CMS001.
METHODS 1. According to the anti-fatigue test, the male Balb/c mice were tested for the exhaustive swimming time, liver starch, urea nitrogen and lactic acid in the blood serum. 2. To research the ultrastructure especially mitochondria and sarcoplasmic reticulum of the fatigued mice and the molecular peptide's effect, electron microscope was used to discover the ultrastructure difference between peptide group and saline group. 3. The MDA, SOD, AST, ALT levels in the blood serum were also detected to find the effects of CMS 008and CMS 001 on the exhaustive mice. 4. The effects of CMS008 and CMS001 on T lymphocyte transformation in vitro were tested to reflect their immune function. 5. Wheel System to Count Mice Activity was designed to count the mice's daily activity. 6. CP 0.1ml(6.0X 109 /ml) was intraperitoneally injected to female C57BL/6 mice to induce immunologically mediated fatigue. What we do is to investigate if CMS 001 had some therapeutic effect on CFS mice.
RESULTS 1. CMS 008 can remarkably prolong exhaustive swimming time (p<0.05). CMS 001 high and low dose groups also can prolong exhaustive swimming time of mice. 2. All the CMS 008 and CMS001 groups can decline the BUN significantly after 90min swimming (p<0.05). 3. CMS 008 high and low dose groups enhance liver starch level significantly contrast to saline group (p<0.05). CMS001 high dose group can enhance liver starch (p<0.05), but the low dose group not. 4. As to lactic acid level in blood serum, it increased after acute short time exercise. All of the experiment groups can inhibit the produce of lactic acid and decrease the increased lactic acid (p<0.05). 5. Electron microscope gave a
direct damage impression of acute long exercise. Skeletal muscle and myocardium's mitochondria and sarcoplasmic reticulum of saline group were damaged by piles and piles of myelin, calcium-overload can also be detected. Mitochondria turned into many vacuoles, sarcoplasmic reticulum lysised. CMS 008 group damaged little at the same time, and pathological changes were minimal to adapt to exercise. 6. After acute exercise, MDA and SOD gained positive findings statistically for the two peptides can decrease MDA and increase SOD contrast to saline group; After exhaustive exercise, AST and ALT levels in the blood were higher than the normal control group, but CMS 008(500ug/kg/d) group was relatively lower than saline group (p<0.05). CMS 001 (20ug/kg/d) group can also decreased AST and ALT levels of blood serum compared with saline group(P< 0.05). 7. CMS 001 can inhibit T lymphocyte transformation^ 0.001), but CMS008 hasn't the immune function.8. Wheel System to Count Mice Activity can count mice's daily activity.
9. Through CP injection, the CFS model mice declined their daily exercise below to 50% of their baseline, and the model control group kept their normal activity, CMS 001 and saline control's activity dropped radically to 20% of the normal activity, and the two groups were in the same percentage (p>0.05).
CONCLUTIONS 1. CMS 008, CMS 001 have anti-fatigue effects on Balb/c mice. 2. Inhabiting protein lysis, protecting skeletal muscle and myocardium, cleaning out free radicals contribute to anti-fatigue of the small peptides. 3. Wheel System to Count Mice Activity can sensitively and precisely counts daily activity of mice, CMS 001 has little therapeutic effect on CFS.
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