【作者】 周忠民；

【导师】 凃晋文；

【作者基本信息】 湖北中医学院 ， 中医内科， 2009， 博士

【摘要】 目的：本研究课题拟通过滋补肝肾方和乙烯雌酚对治疗去卵巢骨质疏松模型大鼠的对比研究，观察去卵巢骨质疏松模型大鼠血清性激素水平FSH、LH、E₂，血清骨钙素（BGP），胰岛素样生长因子-1（IGF-1）水平的变化、脏器指数测定、骨组织病理改变、RT-PCR方法观测IGF-1、OPG-mRNA的表达来探讨滋补肝肾方对去卵巢骨质疏模型大鼠的治疗作用及作用机制。方法：1血液性激素指标测定：摘眼球取血，分离血清，放射免疫法测定血清性激素指标雌二醇（E₂）、卵泡刺激素（FSH）、黄体生成激素（LH）2血液骨代谢生化指标测定：摘眼球取血，分离血清，放射免疫法测定血清骨钙素（BGP）、胰岛素样生长因子-1（IGF-1）。3子宫湿重、脏器指数、骨组织病理改变：将动物断头处死，游离子宫组织，计算脏器指数；取出右侧股骨，以甲醛固定，取股骨头／股骨干处做病理切片，做成石蜡包块，HE染色，光学显微镜下进行病理观察。4骨标本的RT-PCR：断头处死大鼠后完整取出左侧股骨，放入液氮中冷冻，并分装在无菌EP管中密封，以备检测IGF-1、OPG-mRNA水平。结果：1．去势后形成的实验性骨质疏松症大鼠模型，其血清E₂水平明显降低，FSH、LH水平明显升高；血清BGP、IGF-1水平显著降低；子宫湿重和脏器指数明显降低；骨小梁变细，骨皮质变薄，骨密度和面积减少；IGF-1、OPG-mRNA表达水平显著低于正常，出现了典型的骨质疏松症。2．滋补肝肾方可以改善去卵巢骨质疏松模型大鼠的生殖内分泌水平，能显著升高E₂水平，降低FSH、LH水平。3．滋补肝肾方能显著升高去势大鼠体内血清BGP、IGF-1的水平。4．滋补肝肾方对去势大鼠的子宫湿重和脏器指数有升高趋势；减轻去势大鼠的骨质疏松程度。5．滋补肝肾方能上调去势大鼠骨组织IGF-1、OPGmRNA的基因表达。结论：综上所述，滋补肝肾方具有调节去势大鼠的生殖内分泌，改善去势大鼠的血液骨代谢，减轻去势大鼠的骨质疏松程度，上调骨组织IGF-1、OPGmRNA的表达。从多途径、多靶点使去势大鼠引起的生殖内分泌、骨代谢指标及骨组织的变化等接近正常水平，避免了雌激素替代治疗带来的副作用。更多还原

【Abstract】 Objectives: This research subject is plans to treats the ovariectomized rats with the traditional Chinese medicine compound prescription and the ethylene female phenol ,and be observed ovary osteoporosis model big mouse blood serum sex hormone level FSH, LH, E₂, insulin type growth factor - 1 （IGF-1） the level change, the internal organs index determination, the bone tissue pathology change, the RT-PCR method observe IGF-1, the OPG-mRNA expression in bone tissues to discuss the traditional Chinese medicine compound prescription to go to the ovary ossein sparse model big mouse’s therapeutic action and to affect the mechanism.Methods:1. Blood sex hormone target determination: Picks the eyeball to draw blood, the separation blood serum, emits the immunization to determine the blood serum sex hormone : estradiol （E₂）, the graafian follicle stimulin （FSH）, the yellow body production hormone （LH）2. Blood bone metabolism biochemistry target determination: Picks the eyeball to draw blood, the separation blood serum, emits the immunization to determine the blood serum bone calcium element （BGP）, insulin type growth factor - 1 （IGF-1）.3. Womb wet weight, internal organs index, bone tissue pathology change: Beheads the execution the animal, drifts away the womb organization, calculates the internal organs index; Right flank takes out the thighbone, is fixed by the formaldehyde, takes the femoral head/stock backbone place to make the pathological section, makes a paraffin wax package of block, HE dyes, under the optical microscope carries on the pathology observation.4. Bone specimen RT-PCR: Left side beheading executes the big mouse, takes out the thighbone completely, puts in the liquid nitrogen to freeze, and loads separately in the aseptic EP tube seals, prepares examines IGF-1, the OPG-mRNA level.Results1. After been ovariectomized, the big mouse be formed the experimental osteoporosis; the blood serum E₂ level reduces obviously, FSH, the LH level elevates obviously; Blood serum BGP, the IGF-1 level obviously reduces; The womb wet weight and the internal organs index reduce obviously; The bone cerebral cortex thins, the bone density and the area reduce; IGF-1, the OPG-mRNA expression level obviously is lower than normally, presented the model osteoporosis mouse.2. The nutritious hepatorenal traditional Chinese medicine may adjust the reproduction endocrine system to a certain extent, reduces FSH, the LH level, and elevates the E₂ level.3. The traditional Chinese medicine compound prescription can obviously elevate the blood serum BGP, the IGF-1 level in model mouse.4. The traditional Chinese medicine compound prescription can elevates the big mouse’s womb wet weight and the internal organs index; and can reduces the big mouse’s osteoporosis degree.5. The traditional Chinese medicine compound prescription can elevates the IGF-1-mRNA, the OPG-mRNA expression state in bone tissue.Conclusion;The possible mechanism of Bushen Huatan Treatment ofosteoporosis is; Huatan Treatment to inhibit adipogenic differentiation of BMSCs, and Bushen Huatan Treat- ment to promote osteoblastic differentiation of BMSCs.更多还原