耳尖刺血对去卵巢拟阴虚内热大鼠神经内分泌的影响

【作者】 周冰；

【导师】 赵百孝；

【作者基本信息】 北京中医药大学 ， 针灸推拿学， 2007， 硕士

【摘要】 目的本研究旨在通过观察耳尖刺血对去卵巢拟阴虚内热大鼠神经内分泌的影响,来探讨耳尖刺血对围绝经期潮热的疗效及部分机理,为临床应用提供依据和指导。方法雌性Wistar大鼠60只(180～250g),随机分为5组:正常组、假手术组、模型对照组、雌激素对照组、耳尖刺血组。模型对照组、雌激素对照组、耳尖刺血组行双侧卵巢摘除术,假手术组行同样操作,只是不摘除卵巢。手术后第10天,模型组、雌激素对照组、耳尖刺血组采用50%的附子干姜液灌胃造模,正常组、假手术组则灌服等容积蒸馏水。用去卵巢加灌服热性中药的方法复制人工绝经的拟阴虚内热大鼠模型,以己烯雌酚为阳性对照药物,采取耳尖刺血为干预措施,其余组均采用灌服等容积蒸馏水进行对照。用电脑数字式体温计测定大鼠肛温;光镜下观察大鼠阴道脱落上皮细胞角化情况;摘取大鼠子宫,精密测量重量并计算子宫指数;采用放射免疫法测定血清E2(雌二醇)、FSH(促卵泡生成激素)、LH(促黄体生成激素);采用高效液相色谱-电化学检测器测定下丘脑NE(去甲肾上腺素)、5-HT(五羟色胺)、5-HIAA(五羟吲哚乙酸)。比较分析各组观测指标差异性。结果1.大鼠肛温的变化模型组大鼠肛温有上升趋势,在治疗第6天相对正常组、假手术组、雌激素素大鼠肛温显著上升(p<0.05)。雌激素组大鼠肛温在前3次测量中均显著低于模型组(p<0.05)。耳尖刺血组肛温与模型组无明显差异。2.大鼠阴道角化细胞情况正常组、假手术组、雌激素组大鼠涂片均可见大量上皮细胞、角化上皮细胞,模型组与耳尖刺血组则基本以大量白细胞、多核白细胞为主。模型组相对正常组、假手术组阴道上皮细胞角化程度明显下降。雌激素组阴道上皮细胞角化程度高。耳尖刺血组与模型组大鼠阴道上皮细胞角化程度无差异性。3.大鼠子宫重量指数的变化模型组大鼠子宫重量指数与正常组、假手术组相比下降极显著(p<0.001)。雌激素组与模型组相比升高极显著(p<0.001);耳尖刺血组与模型组相比有升高趋势。4.大鼠血清性激素的变化模型组较正常组、假手术组血清E2下降极显著(p<0.01),FSH上升极显著(p<0.01),LH有上升趋势。雌激素组与模型组相比E2有升高趋势,FSH显著下降(p<0.05),LH有下降趋势。耳尖刺血组与模型组相比E2有升高趋势,FSH显著下降(p<0.05),LH有下降趋势。5.大鼠下丘脑神经递质的变化模型组与正常组相比NE显著升高(p<0.05),5-HT、5-HIAA有下降趋势,5-HT/NE比值显著下降(p<0.05)。雌激素组与模型组相比,NE显著降低(p<0.05),5-HT下降极显著(p<0.01),5-HT/NE比值显著下降(p<0.05),5-HIAA/5-HT比值显著上升(p<0.05)。耳尖刺血组与模型组相比,NE显著降低(p<0.05),5-HT下降极显著(p<0.01),5-HIAA上升显著(p<0.05),5-HT/NE比值显著下降(p<0.05),5-HIAA/5-HT比值上升极显著(p<0.01)。结论耳尖刺血有降低肛温、LH的趋势,升高E2的趋势,降低FSH、NE、5-HT的含量,降低5-HT/NE的比值,提高5-HIAA/5-HT比值。耳尖刺血法能够调节去卵巢拟阴虚内热大鼠的神经内分泌功能,对围绝经期潮热有一定的治疗作用。更多还原

【Abstract】 Objective: Through observation of the effects on the neuroendocrine functions of climacteric rat model with interior heat syndrome due to deficiency of yin by ear-apex bloodletting method to investigate its therapeutic effects and mechanism on hot flush and support for the clinical practice.Method: 60 female Wistar rats (180～250g) were randomly divided into 5 groups of normal group, sham-operated group, model group, Diethylstibestrol (DES) group and ear-apex bloodletting(EAB) group. Rats in model group, DES group and EAB group were removed with the ovaries while the sham-operated group with the same operations but without removal of the ovaries. On the tenth day of post-operation, rats in the model group, DES group and EAB group were manipulated with lavage of 50% Fuzi Ganjiang Decoction (Prepared Common Monkshood Daughter Root & Dried Ginger) to prepare interior heat syndrome while the normal group and sham-operated group with lavage of distilled water as contrast. Climacteric rat model with interior heat syndrome due to yin deficiency by method of removal of ovaries and lavage of 50% Fuzi Ganjiang Decoction, were treated with different methods of ear-apex bloodletting, lavage of DES while the control groups with lavage of distilled water. Rat rectal temperature was measured by digital themometor; Rat desquamated Vagina Epithelial Cell was observed under light microscope; Rat uterus was precisely measured after removal and then for calculation of uterus weight index; Rat serum Estradiol (E2), Follicle-stimulating Hormone (FSH) and Luteinizing Hormone (LH) were measured by radio-immunity method; Rat hypothalamus NE, 5-HT and 5-HIAA were measured by High performance Liquid Chromatography with Electrochemical Detection (HPLC-ECD).Result: 1. Rat Rectal Temperature: Rectal temperature in model group had an increasing tendency and obviously increased higher than those in normal group, sham-operated group and DES group (p<0.05). Rat rectal temperature in DES group obviously decreased lower than those in model group (p<0.05). There were no obvious difference between ear-apex bloodletting group and model group. 2. Rat Vagina Epithelial Cell: Vaginal smear results showed a lot of epithelial cells and cuticular epithelial cells in normal group, sham-operated group and DES group while in model group and ear-apex bloodletting group with a lot of leucocytes and polycytes. The extent of cuticularization of the vaginal epithelial cells was obviously lower than that in normal group and sham-operated group while the DES group with higher extent. There were no difference between the EAB group and model group. 3. Rat Uterus Weight Index: Uterus weight index in model group descended extremely remarkable in contrast with normal group and sham-operated group (p<0.001). Index in DES group ascended extremely remarkable in contrast with that of model group (p<0.001). Index in EAB group had an increasing tendency in comparison with that of model group. 4. Rat Serum Hormones: E2 in model group descended obviously than those of normal group and sham-operated group (p<0.01), FSH ascended obviously (p<0.01) and LH had an increasing tendency. In contrast with model group, E2 in DES group and EAB group had an increasing tendency, FSH descended obviously (p<0.05) & LH had a decreasing tendency. 5. Rat Hypothalamus Neurotransmitters: In contrast with normal group, NE in model group obviously increased (p<0.05), ratio of 5-HT/ NE obviously descended (p<0.05) and 5-HT and 5-HIAA had a descending tendency. In contrast with model group, NE and ratio of 5-HT/NE in DES group obviously descended (p<0.05), 5-HT descended significantly (p<0.01) and 5-HIAA/5-HT ascended obviously (p<0.05). In comparison with model group, NE and ratio of 5-HT/NE in EAB group obviously descended (p<0.05), 5-HT descended significantly (p<0.01), 5-HIAA increased obviously (p<0.05) and 5-HIAA/5-HT ascended significantly (p<0.01).Conclusion: Ear-apex bloodletting has functions of lowering rat rectal temperature, LH, FSH, NE, 5-HT & ratio of 5-HT/NE and improving E2 and ratio of 5-HIAA/5-HT. Ear-apex bloodletting method can effect on the neuroendocrine functions of climacteric rat model and has certain effect on treating climacteric syndrome.更多还原