补肾壮骨丸抗骨质疏松的药效学作用及机制探讨

Explore on the pharmacodynamic effect and mechanism of resistance os-teoporosis Bushenzhuanggu Pills

目的探讨北京积水潭医院院内制剂补肾壮骨丸抗骨质疏松的药效学作用及其机制。方法 6个月龄雌性SD大鼠60只,随机分为正常对照组、模型对照组、补肾壮骨丸低(1.2 g/kg)、中(2.4 g/kg)、高(4.8 g/kg)剂量组以及阳性药雷洛昔芬组,每组10只。双侧卵巢摘除术建立大鼠骨质疏松模型,去势手术1周后,补肾壮骨丸低、中、高剂量组及阳性药组分别灌胃给予不同剂量补肾壮骨丸混悬液及雷洛昔芬混悬液治疗12周,正常及模型对照组灌服等体积的生理盐水。分别检测血清碱性磷酸酶(ALP)、抗酒石酸酸性磷酸酶(TRAP)、甲状旁腺激素(PTH)、促甲状腺激素(CT)以及垂体-性腺轴相关激素雌二醇(E2)、睾酮(T)、黄体生成素(LH)、卵泡刺激素(FSH)含量;测定骨密度、骨生物力学性能。结果去势后,模型对照组血清ALP、TRAP、PTH活性显著升高(P<0.05或P<0.01),含量分别为(16.347±2.957)U/100 m L、(27.691±7.042)U/L、(45.829±17.007)pg/m L,CT水平显著下降,含量为(20.469±7.343)pg/m L,全股骨和股骨头骨密度为(0.1859±0.0137)、(0.1871±0.0193)g/cm2,股骨最大载荷为(63.448±17.540)N,最大挠度为(0.5848±0.1595)mm,均较正常对照组显著降低(P<0.05或P<0.01);性激素E2和T含量显著降低(P<0.01),分别为(5.935±1.621)pg/m L、(3.382±1.405)ng/m L,促性腺激素LH和FSH水平明显上调(P<0.05),含量分别为(12.549±2.376)、(15.841±1.896)m U/m L;与模型对照组比较,补肾壮骨丸中、高剂量组ALP含量为(13.619±2.881)、(12.720±3.044)U/100 m L,TRAP含量为(20.904±6.454)、(19.933±5.988)U/L,PTH含量为(26.753±13.013)、(25.630±15.618)pg/m L,三者活性均显著下调(P<0.05或P<0.01);高剂量组CT含量显著升高,为(28.350±7.827)pg/m L,中、高剂量组全股骨密度及高剂量组股骨头密度显著升高,分别为(0.2081±0.0113)、(0.2135±0.0139)、(0.2304±0.0290)g/cm2,高剂量组股骨最大载荷及挠度分别为(81.314±16.413)N、(0.7358±0.1488)mm,较模型组显著升高(P<0.05);高剂量组E2、T显著升高(P<0.05或P<0.01),含量为(8.799±2.204)pg/m L、(5.086±1.668)ng/m L;中、高LH含量分别为(10.740±1.652)、(10.575±0.886)m U/m L,FSH含量分别为(13.244±2.089)、(13.348±2.170)m U/m L,二者水平均显著下调(均P<0.05)。结论补肾壮骨丸可有效纠正骨代谢异常,增加骨密度,增强生物力学性能,其发挥抗骨质疏松作用的部分机制与其调节垂体-性腺轴相关激素水平有关。

Objective To explore the pesticide effect and mechanism of Bushenzhuanggu Pills on osteoporosis rats induced by ovariectomy. Methods Sixty 6-month old female SD rats were randomly divided into six different groups according to body mass： normal control group, model group, Bushenzhuanggu Pill low（1.2 g/kg）, medium（2.4 g/kg） and high dose（4.8 g/kg） groups, while Raloxifene as the positive control drug, 10 rats in each group. All rats received ovariectomy to establish osteoporosis model except those in the normal control group. Rats in treatment groups were treated with intragastric administration of Bushenzhuanggu Pill suspl with different doses or positive drug respectively for twelve weeks. Later the content of alkaline phosphatase（ALP）, tartrate-resistant acid phosphatase（TRAP）, parathyroid hormone（PTH）, osteocalcin（CT）, bone mineral density（BMD）, biomechanical efficiency, pathomorphology were observed. Also the levels of estradiol（E2）, testosterone（T）, luteinizing hormone（LH） and follicle stimulating hormone（FSH） were detected. Results The content of serum ALP, TRAP and PTH were significantly higher（P 〈0.05 or P 〈0.01）, the content was（16.347 ±2.957） U/100 m L,（27.691 ±7.042）U/L,（45.829 ±17.007） pg/m L respectively, while CT level was lower than normal control group by（20.469±7.343） pg/m L. Also reduction in BMD and biomechanical efficiency were observed（P 〈0.05 or P 〈0.01）, the femur and femoral head bone density were（0.1859±0.0137） g/cm2and（0.1871 ±0.0193） g/cm2, then the maximum load of femoral was（63.448 ±17.540） N, the maximum deflection was（0.5848±0.1595） mm. In addition, the content of serum E2 and T was significantly lower（P 0.01）, contents of（5.935±1.621） pg/m L,（3.382±1.405） ng/m L, while the level of LH and FSH was significantly higher in ovariectomy rats（P 〈0.05）,contents of（12.549±2.376）,（15.841±1.896） m U/m L. Compared with model group, ALP content in the middle and high dose group of Bushenzhuanggu Pill were（13.619±2.881） and（12.720±3.044） U/100 m L, TRAP content was（20.904±6.454） and（19.933±5.988） U/L, the PTH content were（26.753±13.013） and（25.630±15.618） pg/m L, the three activities were significantly down regulated（P 〈0.05 or P 〈0.01）. Content of CT in high dose group increased, as（28.350±7.827）pg/m L. The whole femur density in middle and high dose group and the density of the femoral head in high dose group significantly increased; bone density were（0.2081 ±0.0113）,（0.2135 ±0.0139） and（0.2304 ±0.0290） g/cm2 respectively.The maximum load of femoral and radial in high dose group were（81.314±16.413） N and（0.7358±0.1488） mm, these indicators were compared to the model group increased significantly（P 〈0.05）. Also treated group could significantly increase the content of sex hormone, while down-regulate the level of gonadotrophic hormone. The content of E2 and T in high dose group significantly increased（P 〈0.05 or P 〈0.01）, content were（8.799±2.204） pg/m L and（5.086±1.668） ng/m L.The content of LH in middle and high dose group were（10.740 ±1.652） and（10.575 ±0.886） m U/m L respectively（P 〈0.05）, while the content of FSH were（13.244±2.089） and（13.348±2.170） m U/m L respectively（P 〈0.05）, the two levels were significantly lowered. Conclusion Bushenzhuanggu Pill can effectively correct abnormality of bone metabolism,increase bone density, enhance biomechanical properties of bone and reverse pathological changes in bone tissue.The mechanism of its anti-osteoporosis is related to the effect of regulating related hormones levels of thepituitary-gonadal axis.