【作者】 黄宏兴；

【导师】 莫新民；

【作者基本信息】 湖南中医药大学 ， 中医内科学， 2011， 博士

【摘要】 目的：研究线粒体活性变化及骨康方对其调控作用,探讨肌肉-骨骼组织质与量的改变和骨质疏松形成的作用机制。方法：选用6月龄SD大鼠48只,随机分为空白组、模型组、观察组、对照组,各12只。造模2周后,空白组、模型组予蒸馏水,观察组予骨康方对照组予戊酸雌二醇灌胃。12周后,收集血清和左侧股骨、股直肌分别行BMD测量和MPTP、Cyt C、SDH及SOD、MDA检测。选用3月龄新西兰兔4只,随机分为观察组和对照组,各2只。观察组予骨康方、对照组予戊酸雌二醇灌胃,连续7天灌胃后,采血制备含药血浆。选用1日龄SD大鼠培养成骨细胞,成骨细胞分3组培养：空白组不添加任何血浆、观察组加骨康方含药血浆、对照组加戊酸雌二醇含药血浆培养1周后,行MPTP、SDH检测。结果：1.股骨BMD值：模型组低于空白组(P<0.01)；观察组、对照组分别与模型组比较,差异有统计学意义(P<0.05)。2.骨骼肌方面：在MPTP活性方面,空白组、观察组、对照组的OD比值均低于模型组(P<0.05)；各组Cyt C含量变化不大(P>0.05)；在SDH方面,观察组与模型组比较,差异有统计学意义(P<0.05)3.血清SOD和MDA方面：空白组、观察组、对照组与模型组比较,差异均有统计学意义(P<0.05)。4.成骨细胞方面：空白组、观察组的MPTP荧光强度分别与对照组比较,差异有统计学意义(P<0.05)；但SDH含量在各组中的变化不大(P>0.05)。结论：骨康方具有提高去势大鼠BMD的作用；能够有效调控骨骼肌和成骨细胞MPTP的开放,提高线粒体SDH的活力和血清SOD的含量,降低血清MDA的活性,起到预防线粒体损伤的作用,从而抑制细胞凋亡,达到防治骨质疏松症的目的。为从“肾主骨,脾主肌,脾肾相关”角度防治骨质疏松症,筛选以线粒体为作用靶点的药物提供了科学依据。更多还原

【Abstract】 Objective:Analysis the mitochondria activity of muscles-bones in ovariectomized rats and the intervention of GuKang, which to investigate the relationship between the change of qualitative and quantitative of muscle-bone and the formation mechanism of osteoporosis.Method:48 female non-pregnant SD rats(6 months) were randomly divided into 4 groups: normal group(n=12), ovariectomized group(n=12), control group(n=12) and observation group(n=12). After establishing model, the rats of observation group were given the intragastric administration with GuKang. The rats of control group were given estradiol valerate, other groups were given distilled water. After treatment for 12 weeks, the bone mineral density (BMD) and the content of super oxide dismutase(SOD), malondialdehyde(MDA) were detected, mitochondrion were extracted from the muscle tissue that was collected during the operation, and cytochrome c(Cyt C), the mitochondrial permeability transition pore(MPPT) and Succinatedehydrogenase (SDH) were measured. 4 New Zealand rabbits(3 months) were randomly divided into control group(n=2) and observation group(n=2). The observation group was given the intragastric administration with GuKang. The control group was given the intragastric administration with estradiol valerate, After treatment for 1 week, blood sampling was used to separate plasma.1 day old SD rats were used to culture osteoblasts, which were divided into3 groups:normal group, control group and observation group, normal group was not added any drug-containing plasma, observation group was added culture medium of drug-containing plasma of GuKang, control group was added culture medium of drug-containing plasma of estradiol valerate. after 1 week, MPTP、SDH of osteoblasts were measured.Result:1. Femoral BMD:normal group was higher than ovariectomized group (P<0.01); There was a statistically significant difference that the value of observation group, of the control group respectively compared with the ovariectomized group (p<0.05).2. In MPPT、Cyt C and SDH of skeletal muscle, OD ratio of normal group, control group and observation group were lower than ovariectomized group in MPTP(P<0.05). But there was not a statistically significant difference that the content of observation group, of the control group and of normal group respectively compared with the ovariectomized group in Cyt C (P>0.05). There was a statistically significant difference between observation group and ovariectomized group (P<0.05). There was not a statistically significant difference normal group and ovariectomized group(P>0.05).3. In-SOD and MDA, there was a statistically significant difference between normal group and ovariectomized group (P<0.05). There was a statistically significant difference that the value of observation group, of the control group respectively compared with the ovariectomized group (P<0.05).4. In MPTP and SDH of osteoblast, there was a statistically significant difference that the RFU of observation group, of the normal group respectively compared with the control group (P<0.05). But there was not a statistically significant difference that the content of observation group, of normal group respectively compared with the control group in SDH(P>0.05). Conclusion:GuKang could improve BMD of Ovariectomized Rats. Through intervention of GuKang, the permeability of MPTP in skeletal muscle and osteoblast can be controlled. In addition it could improve the content of SDH and reduce the content of MDA, which.can clean up free radicals to inhibit cell apoptosis. These researches could provide the objective reasonable quantification basis for mitochondrion as a therapeutic target from the theory of "kidney controlling bones, spleen controlling muscle and spleen-renal interrelationship" in the treatment of osteoporosis.更多还原