【作者】 张雪竹；

【导师】 孙彦；

【作者基本信息】 天津大学 ， 生物化工， 2009， 博士

【摘要】 以快速老化骨质疏松SAMP6小鼠及其正常对照小鼠SAMR1为研究对象,以中药复方“黄地散”为治疗手段,通过考察血清睾酮(T)、骨钙素(BGP)、血清超氧化物歧化酶(SOD)、丙二醛(MDA)及低密度脂蛋白胆固醇(LDL C)水平、组织钙含量、主动脉钙化情况、股骨生物力学性能、主动脉及股骨中局部骨代谢因子骨形态发生蛋白2(BMP 2)和护骨素(OPG)表达等的变化,观察黄地散对主动脉钙化和骨质疏松症的影响。探讨动脉钙化与骨质疏松的相关性,揭示黄地散对骨质疏松鼠主动脉钙化的调控作用及调控机制。与SAMR1小鼠比较,SAMP6对照组小鼠血清T明显低下;BGP水平明显升高。SAMP6血清SOD水平明显下降,而MDA浓度明显升高;SAMP6血钙、骨钙浓度较低;尿钙、主动脉钙含量较高。形态学显示SAMP6小鼠主动脉中膜平滑肌细胞排列紊乱;中弹力板多处疏松断裂消失;中膜弹性纤维间有细小黑色钙沉积颗粒。SAMP6股骨显微结构破坏,生物力学性能较SAMR1降低。SAMP6对照组主动脉OPG表达下降,BMP 2表达上升;而股骨OPG、BMP 2表达均显著下降。黄地散促使SAMP6小鼠血清T含量明显上升;血清BGP水平明显下降;血SOD上升,血MDA浓度下降。黄地散还对SAMP6异常的组织钙分布有良性调节作用,使血钙、骨钙上升;而尿钙、主动脉钙下降。还可改善SAMP6主动脉结构,减少主动脉钙沉积。黄地散对SAMP6股骨形态计量学、生物力学指标也有明显改善作用,可促进骨形成,抑制骨吸收,增加骨量,提高骨强度。黄地散可对SAMP6主动脉、股骨内骨代谢相关因子OPG、BMP 2的表达有明显良性调节作用,使二者表达趋于正常。骨质疏松小鼠SAMP6主动脉中膜细胞外基质和平滑肌细胞内存在微钙化现象。SAMP6小鼠发生血管钙化并骨质疏松症与性激素分泌下降;脂质过氧化损伤增加;成骨细胞和破骨细胞数目和活性的失调;体内钙平衡遭到破坏,BGP、OPG和BMP 2等局部骨代谢因子表达异常有关。黄地散通过加强性激素分泌及良性调节骨代谢因子OPG、BMP 2表达,既促进骨形成,降低骨吸收,增加骨强度;又抑制了钙盐在主动脉的进一步沉积。因此,黄地散在防治骨质疏松的同时,也提供了对心血管的保护。更多还原

【Abstract】 SAMP6 mice and their normal control mice SAMR1 were enrolled in this study. After 8 weeks of treatment with HuangDi Powder, the serum level of testosterone (T), osteocalcin (BGP), SOD, MDA and LDL C was determined. The calcium content in different tissues, aortic calcification, the mechanical properties of femur, and the expression of local bone metabolism factors (OPG and BMP 2) in arteriae aorta and femur were also assessed after experiment. The therapeutic effect of HuangDi powder on arterial calcification and osteoporosis was studied and the corralation of vascular calcification and osteoporosis was explored. The effect of HuangDi powder on arterial calcification in osteoporotic SAMP6 mice and its possible mechanisms were investigated.Compared to SAMR1 control group, the serum T level of SAMP6 was significantly decreased, whereas the BGP content was obviously increased. The SOD concentration in SAMP6 serum was significantly lower, and the MDA level was notably higher than that in SAMR1. When compared with SAMR1, the calcium content in serum and femur of SAMP6 decreased, while that in urine and arteriae aorta increased obviously. Morphological studies on thoracic aorta showed that the smooth muscle cells in tunica media decreased in amount and arranged disorderly, the elastic lamella were severe disorganized with areas of complete disappearance of the elastic fiber, and numerous calcium deposits were detected within the elastin fibers of tunica media in SAMP6. The microstructure and biomechanical properties of femur in SAMP6 were clearly worse than that in SAMR1. In the aorta of SAMP6, the OPG expression was down regulated and the BMP 2 expression was up regulated, but in the femur of SAMP6, the expression level of both OPG and BMP 2 decreased sharply than that in SAMR1.After the treatment with HuangDi powder, the serum T level in SAMP6 markedly heightened, whereas the OC concentration sharply declined. The increased serum SOD and the decreased MDA contents also were found in SAMP6 mice. HuangDi powder also improved abnormal tissue distribution of calcium to some degree, which induced an increase in calcium content in serum and femur and a decrease in calcium concentration in urine and arteriae aorta of SAMP6. HuangDi Powder could remodel the abnormal architecture of aorta wall and reduce the calcium deposition in thoracic aorta. The bone histomorphometric and mechanical indexes of SAMP6 also improved significantly. HuangDi powder was effective in promoting bone formation, restraining bone resorption, restoring the amount of bone volume and improving bone strength. Moreover, HuangDi powder could also restore the expression of OPG and BMP 2 in thoracic aorta and femur of SAMP6 to normal level.Some calcium deposits were observed in the extracellular matrix and the smooth muscle cells in aortic tunica media of osteoporotic SAMP6. Thoracic aortic calcification and osteoporosis observed in SAMP6 were correlated with the deficiency in testosterone secretion, the increase of lipid peroxidation, the imbalance in numbers and activity of osteoblast and osteoclast, the disorders of calcium balance of organisms and the abnormal expression of local bone metabolism factors (BGP, OPG and BMP 2). HuangDi powder not only promoted bone formation, restrained bone resorption and increased bone strength, but also suppressed calcium deposition in aorta by stimulating testosterone secretion and regulating the expression of OPG and BMP 2. Therefore, HuangDi Powder was effective in reversing osteoporosis of SAMP6 to some degree, and protecting the cardiovascular health at the same time.更多还原