【作者】 颜灿群；

【导师】 陈友香； 夏振信；

【作者基本信息】 武汉大学 ， 内科学， 2004， 硕士

【摘要】 目的：细胞因子在造血、免疫反应和预防感染方面起重要作用。近年来随着免疫学的不断发展，细胞因子在绝经期骨质疏松症发病机制中的作用日益受到重视，尤其是白细胞介素-6(interleukin-6，IL-6)引起了众多研究者的兴趣。大量的临床和实验研究表明这种细胞因子和骨质疏松有着密切关系。其中IL-6被认为是病理状态下调节骨吸收的因子。以往的实验已经证实，具有补肾填精壮骨的补肾壮骨方能够有效改善去卵巢大鼠(模型鼠)的卵巢功能，调节性激素水平，而性激素变化是导致骨质疏松的主要原因，但该药防治骨质疏松的机理不十分明了。因此，本课题在前期工作的基础上，从骨矿密度(BMD)、血脂含量、脏器指数和细胞因子等方面观察各组药物对骨质质疏松大鼠的骨内外影响，并探讨其抗骨质疏松的可能机制，为临床预防和治疗绝经期骨质疏松症提供理论依据。 方法：选用8个月龄雌性Wistar大鼠48只，建立切除卵巢诱发绝经后骨质疏松症的大鼠模型，按体重随机分层分为6组，每组8只，分别以生理盐水，尼尔雌醇，补肾壮骨方大剂量，中剂量，小剂量等药物灌胃，每日一次，共进行12周。12周末麻醉后行双能X线骨密度诊断仪(DEXA)扫描，而后开胸直接从心脏抽血测定血脂(包括低密度脂蛋白(LDL)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白(HDL))，紧接着以，多聚甲醛心脏灌注成功后取大鼠第3腰椎固定、乙二胺四乙酸钠(EDTA)微波脱钙满意后采用免疫组织化学的方法观察IL-6对骨质疏松大鼠中轴骨的影响。 结果： 1．模型组大鼠股骨、腰椎骨及全身的骨密度明显降低(P＜0.05)，补肾壮骨方大、中、小各剂量组和尼尔雌醇组可以预防骨密度的降低。 2．模型组大鼠血清中TG、TC、LDL水平升高，HDL降低(P＜0.05)，各处理组药物均可使TG、TC、LDL水平降低，HDL升高(P＜0.05)。 3．模型组与假手术组比较，子宫与肾上腺重量均明显下降(P＜0.05)；各处理组则上升(P＜0.05)。 4.模型组大鼠切片中，IL一6免疫细胞化学反应呈阳性显示，而各处理组药物中IL一6免疫反应吸收光密度有显著降低(P<0 .05)，阳性细胞白分率明显降低(P<0 .05) 结论:各处理组均能不同程度地提高骨质疏松大鼠的骨密度，调节血脂水平，防止子宫和肾上腺萎缩以及降低IL一6在中轴骨中的表达，综合比较以补肾壮骨方大、中剂量组和尼尔雌醇组的效果最好。补肾壮骨方可以适合于绝经后妇女长期服用，以预防与绝经有关的骨质疏松症，并能保护心血管等更多还原

【Abstract】 Objects Cytokines are essential for hematopoiesis and immune responses, and they play a key role in the defense against infection. In recent years , with the development of immunology, the role of cytokines have been increasingly concerned in the pathogenesis of postmenopausal, especially interleukin-6 ( IL-6 ) is an important proinflammatory cytokines. It has been demonstrated that IL-6 is closely related to osteoporosis by enormous clinical and experimental studies. IL-6 is regarded as a cytokine of regulating bone absorption. It has been demonstrated that Bushen Zhuanggu Prescription could effectively improve the function of ovary of ovrariectomized rats. It also can regulate the level of gonadal hormone. The change of level in gonadal hormone is a main cause which leads to osteoporosis .However, the machanism of Bushen Zhuanggu Prescription on osteoporosis have not been clearly claritied. The approach used here was based on the former fundermental research. We observed effect of different dose Bushen Zhuanggu Prescription groups on bone metabolism bood lipid cytokine and organism index. By using the immunohistochemistry technique, IL-6 was tested to study the mechanism of the Bushen Zhuanggu Prescription prophylaxis and treatment PMO. By conducting experiment, effects of different treatments were compared to see whether difference exists in order to provide theoretical evidence for clinical manifestation prophylaxis and treatment.Methods In this experiment, 48 unfertilized female rats were chosen as the experimental objects ,whose ages were all eight -month-old and weight from 242 to 318 grams. By doing oophorectomy operations on 40 rats, fake operations on 8 rats ,the rats were randomly classified into five groups according to their weights , 8 rats in each.They were ovariectomized group, nilestrial group and kidney tonifying herbs ( Bushen Zhuanggu Prescription, BSZG) in different doses. Both BSZG group and nilestrial group were conducted medicine treatments for 7 days after operation. The treatment period last for 12 weeks . After the last treatment, BMD ( Bone Mineral Density) was tested by Dural-EnergyX-ray Absorptiometry ( DEXA ) . And then serum lipid was measured after centrifugation. The effect of IL-6 on the 3rd lumbar vertebrae of ovariectomized rats was observed by immunohistochemistry after heart perfusion localization and decalcification by edetic acid (EDTA). Results1. Level of BMD of lumbar veterbra and thighbone in ovariectomized rats decreased ( P<0.01 ) BSZGP could preventing losing of bone mineral.2 TC, TG, LDL levels in Model rats increased, HDL serum decreased ( P0.01 ) , while other groups were opposite results.3 Model rats’ weight of both uterus and adrenal is obviously lighter than sham operation and other groups.4 Model rats’ immune celluar chemical response to IL-6 was positive, while other groups all decreased in both O.D. and positive cell percentage.Conclusion All the experimental groups had some therapeutic effect on increasing BMD,regulating serum lipid and decreasing the express of IL-6 in the axial bone. By systematic statistics , it was found that the result of nilestrial .moderate and high doses of BSZGP were the best. BSZG could be taken by postmenopausal women for long time to prevent osteoporosis which is related to PMO, it could also protect heart by regulating serum lipid.更多还原