铁皮石斛水提物对糖尿病小鼠血糖和肠道菌群的影响

发布时间：2018-01-12 05:17

本文关键词：铁皮石斛水提物对糖尿病小鼠血糖和肠道菌群的影响　出处：《大连医科大学》2017年硕士论文　论文类型：学位论文

更多相关文章： 铁皮石斛 糖尿病 降血糖 肠道菌群

【摘要】：目的:研究人工培育的铁皮石斛水提取物,对链脲霉素(STZ)所致的高血糖小鼠血糖、血脂、机体的脂肪和肌肉组织成分比例和肝脏抗氧化能力的影响,分析对肠道菌群影响。通过多种途径分析高血糖对机体的影响,以及铁皮石斛水提物对机体的保护作用和可能的机制。方法:腹腔注射STZ,制备小鼠糖尿病模型。实验分为铁皮石斛水提物高、中、低剂量组、正常对照组、模型对照组、阳性药物组。观察实验动物的一般情况,记录小鼠体重,用核磁共振仪测定小鼠机体组织成分比例,用血糖试纸测空腹血糖值,血清TC、TG、HDL值按照试剂盒说明检测,利用分光光度计测肝脏SOD含量(黄嘌呤氧化酶法)、TBA法测MDA含量,通过PCR-DGGE凝胶电泳,quantity one软件分析小鼠肠道菌群多样性。结果:(1)正常对照组小鼠在整个实验过程中,生长情况良好,没有死亡现象。成模小鼠精神不振、反应慢、不爱活动,每日喝水量增加,排尿次数明显增加、体重明显下降(P0.05)、毛发松驰无光泽、好激怒、粪便变小,变黄且发干,模型对照组存活状态最差,存活率仅为50%。石斛水提物三个剂量组和阳性药物组小鼠的一般状态均明显改善,毛色和精神状态有所恢复,饮水量和尿量减少,粪便着色变深,存活率分别为80%、60%、60%、80%。模型对照组机体脂肪与肌肉的比例:正常对照组小鼠,随体重的增加,脂肪组织明显增加,明显高于其他各组,脂肪组织所占比例肌肉组织。模型对照组小鼠,随体重的下降,脂肪组织也减少,与其他各组相比有很大差别,脂肪组织所占比例肌肉组织。石斛水提物三个剂量组及阳性药物组,脂肪组织的变化与体重的变化一致,不论是脂肪组织的占比例还是肌肉组织所占比例均高于模型组,并且肌肉组织所占比例高于正常对照组。(2)与正常对照组比较,模型对照组血糖水平显著上升,在建模第28d后开始下降,3个石斛水提物组血糖水平在第7d起与模型对照组比有所减少,14~28d减少更加明显,差异有显著性(p0.01),这一结果与阳性药物组的血糖的变化一致,并且各组彼此没有显著差异(p0.05)。(3)建模28d后,与正常对照组比较,模型对照组小鼠血清tc、tg含量明显高于正常对照组小鼠(p0.05),hdl低于正常对照组(p0.05)。与模型对照组比较,石斛水提物三个剂量组和阳性药物组的血清tc含量明显减少(p0.05),hdl含量明显增加(p0.05)。石斛低剂量组、阳性药物组tg含量低于模型对照组(p0.05),石斛中、高剂量组的血清tg含量高于模型对照组(p0.05)。(4)与正常对照组比较,模型对照组小鼠肝组织sod活性显著降低(p0.05),mda水平显著上升(p0.01)。与模型对照组比,三个剂量石斛水提物的肝组织sod活性均明显升高、mda含量均减少(p0.05);阳性药物组sod活力有所增加(p0.05),但是mda含量高于模型对照组小鼠。(5)与正常对照组比较,阳性药物组和模型对照组粪便中菌群条带模糊不清,细菌表达量较低。石斛水提物中、高剂量组菌群多样性好且表达量较高。聚类分析结果表明,石斛水提物低、中剂量组为一簇,菌群相似度最高,石斛高剂量组、模型对照组,阳性药物组和正常对照组为一簇,菌群相似度高,其中阳性药物组与正常对照组菌群高度相似,石斛水提物治疗组中以高剂量组的菌群与正常对照组最为相似。以上结果表明,铁皮石斛水提物中、高剂量均能改善糖尿病小鼠的肠道菌群数量和多样性,在提高菌群数量上比阳性药物作用效果好。结论:(1)石斛水提物能够改善糖尿病小鼠机体的健康情况和体重急剧下降的并发症;(2)石斛水提物对stz诱导的糖尿病小鼠有显著的降血糖功效,和阳性药物(盐酸二甲基双胍)效果相差不多;(3)石斛水提物能调节stz糖尿病小鼠机体脂肪与肌肉组织的比例,提示其可能调节了机体脂代谢,从而影响血糖水平;(4)石斛水提物能够调节糖尿病导致的tc、tg升高、hdl降低,改善糖尿病小鼠的血脂混乱;(5)石斛水提物能够通过升高糖尿病小鼠肝组织sod活性、降低mda水平,来增强肝脏抗氧化能力;(5)石斛水提物能够调节糖尿病所导致小鼠肠道菌群失调、菌群数量减少。综上所述,铁皮石斛水提物通过调节机体血脂、肝脏抗氧化能力和肠道菌群,改善了糖尿病小鼠的脂代谢,增强机体免疫能力,进而间接调节了血糖水平的稳定。所以,人工培育的铁皮石斛可以作为治疗糖尿病的辅助药物,能够降低血糖水平及改善糖尿病所导致的部分并发症,为未来糖尿病的预防和治疗提供新思路。
[Abstract]:Objective: To study the artificial cultivation of Dendrobium candidum water extract, on streptozotocin (STZ) hyperglycemia mice induced by glucose, lipid, effect of adipose and muscle composition of body and liver antioxidant ability, analysis of effect on the intestinal flora. Through a variety of ways to analysis the effects of high blood glucose on the body, and Dendrobium the water extract of protective effect on the body and the possible mechanism. Methods: intraperitoneal injection of STZ, the preparation of the diabetic mouse model. The experiment was divided into Dendrobium candidum water extract high, low dose group, normal control group, model control group, positive drug group. General observation of experimental animal, records of body weight of mice determination of body composition of mice, using nuclear magnetic resonance instrument, with blood glucose test strips, fasting blood glucose, serum TC, TG, HDL value detection in accordance with the kit, using the spectrophotometer to measure the content of liver SOD (xanthine oxidase method), TBA method To measure the content of MDA, PCR-DGGE by gel electrophoresis, analysis of mouse intestinal flora diversity of quantity one software. Results: (1) normal control mice during the whole experiment, growth is good, there is no death phenomenon. Model mice Jingshenbuzhen, slow response, not love activities, daily water quantity increased, a significant increase in the number of urination body weight, decreased significantly (P0.05), hair loose without luster, good anger, stool becomes small, yellow and dry, the model control group survived the worst state, the general state of survival rate is only 50%. from aqueous extracts of three dose group and positive drug group mice were significantly improved, the colour and the mental state recovery, water intake and urine volume decreased, fecal staining and survival rate were 80%, 60%, 60%, the proportion of 80%. in model control group, body fat and muscle of the mice in normal control group, with the increasing of body weight, fat tissue increased significantly, significantly higher than the other In each group, the proportion of adipose tissue, muscle tissue. The model control group, with the decrease of body weight, fat tissue also decreased compared with other groups, there is a great difference between the proportion of adipose tissue, muscle tissue. The water extract of Dendrobium three dose group and positive drug group, consistent changes in adipose tissue and body weight. Whether the proportion of adipose tissue or muscle tissue proportion is higher than the model group, and muscle tissue proportion is higher than the normal control group. (2) compared with the normal control group, model control group, blood glucose levels increased significantly in the model, 28d began to decline after 3 d. water extraction group blood glucose level in the 7d and the model control group decreased, 14~28d decreased more obviously, there was significant difference (P0.01) changes, the result was in line with the positive drug group the blood sugar level, and the groups had no significant difference each other (P0.05). (3) modeling 28 After D, compared with normal control group, model control group, serum TC, TG were higher than that of normal control mice (P0.05), HDL lower than the normal control group (P0.05). Compared with the model group, the serum TC content of water extract of Dendrobium three dose group and positive drug group decreased significantly (P0.05) and the content of HDL increased significantly (P0.05). From the low dose group, positive drug group TG was lower than that of model group (P0.05), Dendrobium, the content of serum TG in high dose group was higher than that of the model control group (P0.05). (4) compared with the normal control group, model control group of mice liver tissue SOD activity decreased significantly (P0.05), the level of MDA increased significantly (P0.01). Compared with model control group, three dose of water extract of Dendrobium liver SOD activity were significantly increased, MDA content decreased (P0.05); the positive drug group increased the activity of SOD (P0.05), but the content of MDA was higher than the model control group (5 mice. With normal) According to group, positive drug group and model control group of fecal bacteria with blurred, bacterial expression is relatively low. The water extract of Dendrobium in high dose group, the bacterial diversity and high expression level. The results of cluster analysis showed that Shi Hushui extract low, middle dose group as a cluster, flora similarity the highest Dendrobium high dose group, model control group, positive drug group and normal control group as a cluster of bacteria, high similarity, the positive drug group and normal control group of bacteria was highly similar to that of Dendrobium water extract in the treatment group with the flora of high dose group and normal control group. The above results most similar show that the Dendrobium candidum water extract and intestinal microflora in high dose could improve diabetic mice, increase in number of bacteria than the positive drug effect. Conclusion: (1) Shi Hushui extract can improve diabetic mice body health and weight urgently Play down the complications; (2) Shi Hushui extract had a significant hypoglycemic effect in STZ induced diabetic mice, and positive drug (two methyl guanidine hydrochloride) effect is not much; (3) Shi Hushui extract can regulate the STZ diabetic mice body fat and muscle tissue proportion, suggesting that it may regulate the body lipid metabolism, thus affecting the blood glucose level; (4) Shi Hushui extract can regulate the diabetes induced TC, TG increased, HDL decreased, improve blood lipid of diabetic mice confusion; (5) Shi Hushui extract can increase the activity of SOD in liver tissue of diabetic mice, decrease the level of MDA, to enhance the antioxidant capacity of the liver; (5) Shi Hushui extract can regulate diabetes caused by intestinal flora of mice, the number of bacteria decreased. In summary, the water extract of Dendrobium officinale by regulating blood lipid, antioxidant capacity of liver and intestinal microflora, improve lipid metabolism in diabetic mice Thanks, enhance the immune ability, and indirectly regulate the stability of blood glucose level. Therefore, the artificial cultivation of Dendrobium candidum can be used as auxiliary medicine for treating diabetes, can reduce the level of blood glucose and improve complications due to diabetes, for future diabetes prevention and treatment to provide new ideas.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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