【作者】 牛放；

【导师】 曲晓波；

【作者基本信息】 长春中医药大学 ， 药理学， 2011， 硕士

【摘要】 目的:探讨梅花鹿鹿角脱盘水溶性总蛋白的抗疲劳作用及治疗乳腺增生作用;研究梅花鹿鹿角脱盘胶原蛋白的抗炎、镇痛作用。方法:1样品制备采用缓冲溶液浸提和中空纤维膜分离系统制备梅花鹿鹿角脱盘水溶性总蛋白;应用加热提取和酶解技术制备梅花鹿鹿角脱盘胶原蛋白。2抗疲劳作用选用雄性小鼠分为三个剂量组和空白对照组,灌胃给药不同剂量的梅花鹿鹿角脱盘水溶性总蛋白30d后,测定小鼠力竭游泳时间、血乳酸含量、肝糖原储备以及血清尿素氮等指标。3治疗乳腺增生作用大鼠注射苯甲酸雌二醇,连续25d,再注射黄体酮注射液5d复制大鼠乳腺增生模型。应用鹿角脱盘水溶性总蛋白(高、中、低3个剂量组)腹腔注射给药30d,测定第2对乳房高度及直径,采血测定血清中雌激素和孕激素水平,取胸腺、脾脏、子宫、卵巢称重,计算脏器系数;并将胸前第2对乳腺、子宫及卵巢置于10%甲醛溶液中固定,做病理组织检查。4抗炎作用通过小鼠足肿胀、耳廓肿胀的实验研究梅花鹿鹿角脱盘胶原蛋白的抗炎作用。5镇痛作用采用热板法和醋酸扭体法研究梅花鹿鹿角脱盘胶原蛋白的镇痛作用。结果:1采用缓冲溶液浸提和中空纤维膜分离系统制备梅花鹿鹿角脱盘水溶性总蛋白,收率为1%,蛋白含量为66.29%;采用水煮和酶解法制备梅花鹿鹿角脱盘胶原蛋白,收率为22.31%,蛋白含量为66.32%。2与对照组比较,鹿角脱盘水溶性总蛋白组小鼠负重游泳时间延长,血乳酸水平降低,肝糖原和血清尿素氮含量与空白组无明显差异。3鹿角脱盘水溶性总蛋白可改善乳腺增生病大鼠乳腺的组织形态学结构,缩小乳房大小和乳腺体积,减少乳腺小叶腺泡数、腺泡腔、分泌物,并可使血清雌激素水平降低。4梅花鹿鹿角脱盘胶原蛋白能缓解角叉菜胶引起的足肿胀并抑制二甲苯引起的小鼠耳廓肿胀。5梅花鹿鹿角脱盘胶原蛋白可提高热板实验小鼠的痛域,并能缓解醋酸引起的小鼠扭体反应次数,延长疼痛潜伏期。结论:采用中空纤维膜分离法和酶解法分别制备梅花鹿鹿角脱盘水溶性总蛋白和梅花鹿鹿角脱盘胶原蛋白各50g。应用此方法样品提取率高,损失少,且可保持样品的生物活性。方便易行,应用领域广,可应用于大规模工业生产。梅花鹿鹿角脱盘水溶性总蛋白具有抗疲劳的作用,并能通过调节乳腺增生大鼠的性激素水平,改善增生乳腺的形态学变化而有效地对抗雌激素引起的大鼠乳腺增生。梅花鹿鹿角脱盘胶原蛋白具有一定的抗炎镇痛作用,对风湿、类风湿性关节炎、风湿骨痛的治疗具有广阔的前景,为以鹿角脱盘为基源的新药开发提供了科学依据。更多还原

【Abstract】 Purpose: Discussion the anti-fatigue and hyperplasia effect of Sika antlerbase water-soluble protein; Research the effect of Sika antlerbase collagen on the anti-inflammatory and analgesic.Methods:1 Sample preparationExtract Sika antlerbase water-soluble protein by buffer solution and hollow fiber membrane separation system;get Sika antlerbase collagen by heating and enzyme technology.2 The effect of anti-fatigueMice were divided into 3 dose groups and control group,different doses of Sika antlerbase extraction were applied to mice.Loaded swimming time,blood lactic acid,liver hepatin and blood urea were detemined after 30 days.3 The treatment of hyperplasiaRats inject estradiol benzoate 25 days,then inject progesterone 5 days continue.Measure height and diameter of the second pair of nipple,determine estrogen and progesterone in serum,take thymus,spleen,uterus,ovarian and calculation the organ coefficient,at the same time,fixed the organs in 10%formaldehyde solution to do pathology organization test.4 The effect of anti-inflammatoryThe experiments of anti-inflammatory adopt paw swelling model and ear edema model of mice.5 The effect of analgesicAnalgesia use hot-plate test and acetic acid writhing method.Results:1 Extract Sika antlerbase water-soluble protein by buffer solution and hollow fiber membrane separation system,extraction rate is 1%,protein content is 66.29%;get Sika antlerbase collagen by heating and enzyme technology,extraction rate is 22.31%,protein content is 66.32%.2 The loaded swimming time was prolonged,blood lactic acid level was decreased,but liver hepatin and blood urea level have no significantly difference compared with the control group.3 Sika antlerbase protein can improve the morphology structure of hyperplasia rats, narrow the size and volume of nipple and breast,reduct the quantity of lobular acinar and secretion,at the same time, make the estrogen level in serum lower.4 Sika antlerbase collagen can ease the carrageenan-induced paw swelling and the xylene-induced ear edema.5 Sika antlerbase collagen can improve the pain threshold of the hot-plate test, reduce intra-abdominal inflammation caused by acetic acid exudation, prolong the incubation period pain.Conclusion:Get Sika antlerbase water-soluble protein and Sika antlerbase collagen 50g each by hollow fiber membrane separation system and enzyme technology.We have high extraction rate and less loss by use these methods,at the same time maintain the biological activity of sample.Facilitate the easy,wide application field,can be applied to large scale industrial production.Sika antlerbase protein have effect of anti-fatigue,at the same time,can improve morphological changes of breast by regulation the sex hormones of hyperplasia rats,to effective against rats hyperplasia caused by estrogen.Sika antlerbase collagen have some effect of anti-inflammatory and analgesic,have broad prospects on rheumatoid rheumatoid arthritis and rheumatism,provide scientific basis as new drugs to sources of Sika antlerbase.更多还原