藏药二十五味鬼臼丸通过OPG/RANKL/RANK通路对去卵巢骨质疏松大鼠骨代谢的干预作用

Intervention effect of Tibetan medicine Ershiwuwei Guijiu Pills on bone metabolism in ovariectomized osteoporosis rats via OPG/RANKL/RANK pathway

目的:探究藏药二十五味鬼臼丸通过OPG/RANKL/RANK通路对去卵巢骨质疏松大鼠骨代谢的干预作用。方法:建立去卵巢骨质疏松模型,术后2周开始给药,模型组、假手术组灌胃蒸馏水,雌二醇组灌胃0.1 mg/kg戊酸雌二醇片混悬液,二十五味鬼臼丸低、中、高剂量组分别灌胃200、400、800 mg/kg二十五味鬼臼丸混悬液,1次/d,连续12周。给药后每周统计体质量,HE染色观察股骨和腰椎形态,ELISA法检测大鼠血清骨形成标志物[骨钙素(BGP)、Ⅰ型前胶原氨基端原肽(PⅠNP)、骨特异性碱性磷酸酶(BALP)]和血清骨吸收标志物[Ⅰ型胶原C端肽(CTX-Ⅰ)、抗酒石酸酸性磷酸酶5b(TRAP5b)]水平,RT-PCR检测股骨中骨保护素(OPG)、核因子κB受体活化因子配体(RANKL)、核因子κB受体活化因子(RANK)mRNA的表达水平,Western Blot检测股骨中OPG、RANKL蛋白水平。结果:与模型组比较,二十五味鬼臼丸高剂量组能够显著降低大鼠的体质量和血清CTX-Ⅰ、TRAP5b水平(P<0.01),显著降低股骨RANKL、RANK mRNA及RANKL蛋白的表达(P<0.05),显著提高大鼠血清BGP、PⅠNP、BALP水平和股骨OPG mRNA、OPG蛋白的表达(P<0.01,P<0.05)。结论:藏药二十五味鬼臼丸通过调节OPG/RANKL/RANK信号通路对去卵巢骨质疏松大鼠骨代谢发挥调控作用。

Objective:To explore the intervention effect of Tibetan medicine Ershiwuwei Guijiu Pills on bone metabolism in ovariectomized osteoporosis rats through OPG/RANKL/RANK pathway.Methods:The SD rat model of ovariectomized osteoporosis was established,and the drug was administered two weeks after the operation.The model group and the sham operation group were given distilled water by gavage,Estradiol group was given O.1 mg/kg estradiol valerate tablets by gavage.Ershiwuwei Guijiu Pills low,medium and high dose group were respectively given 200,400,800 mg/kg Ershiwuwei Guijiu Pills once a day,for 12 consecutive weeks.After administration,the body weight was counted every week.Hematoxylin-eosin(HE)staining was used to observe the morphology of femur and lumbar spine.ELISA was used to detect the levels of the serum bone formation marker[osteocalcin(BGP),procollagenⅠN-terminal propeptide(PⅠNP),bone alkaline phosphatase(BALP)]and the levels of serum bone resorption markers[C-telopeptide of typeⅠcollagen(CTX-Ⅰ),tartrate-resistant acid phosphatase 5b(TRAcp5B)],RT-PCR was used to detect the expression level of osteoprotegerin(OPG),receptor activator of the nuclear factor-xB(RANK)and receptor activator of nuclear factor-kB ligand(RANKL)mRNA in the femur,and the protein levels of OPG and RANKL in the femur were detected by Western Blot.Results:Compared with model group,Ershiwuwei Guijiu Pills high dose group could significantly reduce the body weight,serum CTX-Ⅰand TRAP5b levels(P<0.01)and femoral RANKL,RANK mRNA and RANKL protein expressions in rats(P<0.05),Ershiwuwei Guijiu Pills could significantly increased serum BGP,P I NP,BALP levels,and the femoral OPG mRNA and OPG protein expressions in rats(P<0.01,P<0.05).Conclusion:Tibetan medicine Ershiwuwei Guijiu Pills regulates bone metabolism in ovariectomized osteoporosis rats by regulating OPG/RANKL/RANK signaling pathway.