电针促脂肪代谢的腺苷受体机制

发布时间：2017-12-28 23:17

本文关键词：电针促脂肪代谢的腺苷受体机制　出处：《南京中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:观察电针对高脂饮食诱导肥胖(diet induced obesity,DIO)小鼠白色脂肪组织(WAT)中腺苷受体的影响,探讨腺苷受体在电针减肥中的作用机制。方法:将63只3周龄雄性C57BL/6小鼠随机分为普食组(21只)和高脂组(42只),分别给予普通饮食和高脂饮食喂养12周后,以体质量超过正常小鼠平均体质量的20%作为肥胖模型标准,并最终将小鼠随机分为正常对照组(8只)、正常电针组(13只)、模型对照组(9只)、模型电针组(14只)。两个治疗组均电针"后三里"和"内庭"穴,每周治疗6次,共电针4周。测量小鼠体质量和附睾脂肪组织质量,计算小鼠体重变化以及脂肪组织脂体比;用代谢笼监测小鼠热量释放、摄食、活动量(XT+YT)和总移动路程;HE染色评价脂肪细胞的体积;用qPCR技术检测脂肪组织中A1R、A2AR、A2BR、A3R的mRNA表达以及棕色化相关基因Tbx1、Tmem26的表达;用Western Blot检测脂肪组织中腺苷受体、PRDM16、PGC-1α、UCP1、pTH、pHSL 的蛋白表达。结果:实验一:1.与普食组相比,高脂喂养12周能明显增加小鼠体质量(P0.01),其中,体质量超过普食组20%的小鼠有23只(占54.76%)。2.治疗结束后,与正常对照组相比,正常电针组小鼠体质量和附睾脂肪组织(Epi-WAT)脂体比下降(P0.05),模型对照组体质量依然维持在较高状态(P0.05)、体质量变化值差异有统计学意义(P0.01)、Epi-WAT质量和脂体比升高(P0.01,P0.05);与模型对照组相比,模型电针组体质量、Epi-WAT脂体比均降低(P0.05)、体质量变化值增大(P0.01)。3.代谢笼:与正常对照组相比,模型对照组VO2水平下降(P0.05)、呼吸交换率(Respiratory exchange rate,RER)升高(P0.05)、摄食量升高(P0.05)、活动量(XT+YT)和总移动路程无显著差异(P0.05);与模型对照组相比,模型电针组H1、H2、V02、VC02水平均升高(P0.05),RER无统计学差异(P0.05),摄食量下降(P0.05),XT+YT、总移动路程无显著差异(P0.05)。4.HE染色:与正常对照组相比,正常电针组附睾脂肪细胞面积减小(P0.05),模型对照组面积增大(P0.01);与模型对照组相比,模型电针组面积减小(P0.01)。实验二:1.A1RmRNA在四组中表达的差异无统计学意义(P0.05);与正常对照组相比,正常电针组A2AR和A2BR蛋白表达上调(P0.05,P0.01),模型对照组A3R mRNA的表达降低(P0.01)、A2AR蛋白表达降低(P0.05)、A2BR蛋白表达差异不明显(P0.05);与模型对照组相比,模型电针组A2AR、A2BR mRNA表达升高(P0.05),A2AR和A2BR蛋白表达升高(P0.01)。2.与正常对照组相比,模型对照组Tbx1、Tmem26 mRNA表达降低(P0.05),BAT中PRDM16蛋白水平下降(P0.05);与模型对照组相比,模型电针组Tbx1、Tmem26mRNA水平上调(P0.05),BAT中PRDM16、UCP1蛋白水平上调(P0.01),Epi-WAT 中 PRDM16、PGC-1α、UCP1 的蛋白表达升高(P0.05,P0.05,P0.01)。3.与正常对照组相比,正常电针组pTH蛋白表达升高(P0.05),模型对照组pTH、pHSL蛋白水平下降(P0.01,P0.05);与模型对照组相比,模型电针组pTH、pHSL蛋白水平均升高(P0.01)。结论:电针可能通过激活自主神经系统,调节WAT中A2AR、A2BR的表达,促进脂肪代谢和能量消耗,这可能是电针减肥的部分机制。
[Abstract]:Objective: To observe the effect of Electroacupuncture on adenosine receptor in white adipose tissue (WAT) of diet induced obesity (DIO) mice induced by high-fat diet, and to explore the mechanism of adenosine receptor in reducing weight by electroacupuncture. Methods: 63 3 week old male C57BL/6 mice were randomly divided into normal control group (21 rats) and high-fat group (42 rats), which were fed with normal diet and high-fat diet for 12 weeks, with the body mass of more than 20% of the average body weight of normal mice as obesity model standard, and finally the mice were divided into normal control group (8 rats), normal acupuncture group (13 rats), model group (9 rats), model group (14 rats) electroacupuncture. The two treatment groups were treated with electroacupuncture "after three li" and "inner court", 6 times a week for 4 weeks. Measurement of mice body weight and epididymal adipose tissue mass, calculate the weight changes of mice adipose tissue and body fat ratio; release, using metabolic cages of mice feeding and activity monitoring heat capacity (XT+YT) and the total moving distance; HE staining was used to evaluate the size of fat cells; using qPCR to detect the fat tissue of A1R, A2AR, A2BR, A3R the expression of mRNA and browning related gene Tbx1 and Tmem26 expression; expression of adenosine Blot receptors in the detection of adipose tissue Western, PRDM16, PGC-1, UCP1, pTH, alpha pHSL protein. Results: Experiment 1: 1., compared with the common food group, high fat diet can significantly increase body weight (P0.01) in 12 weeks, and the body weight of the mice is 20% higher than that of the common food group (23) (54.76%). 2. after the end of treatment, compared with the normal control group, normal acupuncture group of mice body weight and epididymal adipose tissue (Epi-WAT) decreased body fat (P0.05), model control group, the body mass is still maintained at a high state (P0.05), the mass variation value difference was statistically significant (P0.01), Epi-WAT quality and ratio of fat. Increased (P0.01, P0.05); compared with the model group, EA group model body mass, fat body were lower than Epi-WAT (P0.05), the changes of body weight increases (P0.01). 3. metabolic cages: compared with normal control group, model control group, VO2 decreased (P0.05), respiratory exchange ratio (Respiratory exchange rate, RER) (P0.05), increased food intake increased (P0.05), activity (XT+YT) and no significant difference between the total mobile distance (P0.05); compared with the model control group model H1, H2, V02 EA group, the level of VC02 increased (P0.05), RER (P0.05), there was no significant difference in food intake decreased (P0.05), XT+YT had no significant difference, the total mobile distance (P0.05). 4.HE staining: compared with the normal control group, the area of epididymal adipocytes in the normal electroacupuncture group decreased (P0.05), and the area in the model control group increased (P0.01). Compared with the model control group, the area of the EA group decreased (P0.01). Experiment two: there was no significant difference in the expression of 1.A1RmRNA in the four groups (P0.05); compared with the normal control group, normal acupuncture group of A2AR and A2BR protein expression (P0.05, P0.01), model control group A3R expression decreased mRNA (P0.01), lower expression of A2AR (P0.05) and A2BR protein expression difference obviously (P0.05); compared with the model group, the increased expression of A2AR, A2BR, mRNA model EA group (P0.05), the expression of A2AR and A2BR protein level (P0.01). 2. compared with normal control group, model control group Tbx1, Tmem26 decreased expression of mRNA (P0.05), PRDM16 protein level in BAT decreased (P0.05); compared with the model control group, model group, Tbx1 EA increased level of Tmem26mRNA (P0.05), the level of PRDM16 and UCP1 protein in BAT (P0.01), increased expression of Epi-WAT in PRDM16 PGC-1, alpha, UCP1 protein level (P0.05, P0.05, P0.01). 3., compared with the normal control group, the expression of pTH protein in the normal electroacupuncture group increased (P0.05), and the pTH and pHSL protein levels in the model control group decreased (P0.01, P0.05). Compared with the model control group, the pTH and pHSL protein levels in the EA group increased (P0.01). Conclusion: EA may regulate the expression of A2AR and A2BR in WAT and promote lipid metabolism and energy consumption by activating the autonomic nervous system, which may be a part of the mechanism of electro acupuncture for weight loss.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245

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