益肾健骨膏对去卵巢骨质疏松大鼠骨代谢的影响

Effect of Yishenjiangu Cream Formula on bone metabolism in ovariectomized osteoporosis rats

目的探讨益肾健骨膏对去卵巢骨质疏松大鼠骨代谢的影响及可能机制。方法50只雌性SD大鼠被随机分为假手术组、模型组、高剂量膏方组、低剂量膏方组、阳性药物组,每组10只。假手术组、模型组给予10 mL/(kg·d)生理盐水灌胃,低、高剂量膏方组分别给予2 g/(kg·d)、13 g/(kg·d)益肾健骨膏灌胃,阳性药物组灌服6.25 mg/(kg·w)阿仑膦酸钠维D3。治疗12周后检测大鼠腰椎骨密度和腰椎最大压缩载荷,通过Elisa检测血清骨代谢指标抗酒石酸酸性磷酸酶(TRACP)、骨保护素(OPG)、Ⅰ型前胶原氨基端原肽(PINP)、Ⅰ型胶原交联羧基端肽(CTX-Ⅰ),RT-PCR检测腰椎骨组织OPG、RANKL、RANK、TRAF6的mRNA表达,RT-PCR及Western blot检测腰椎Notch1、Notch2、Jagged1、Jagged2的基因及蛋白表达。结果益肾健骨膏治疗12周后,与模型组比较,高剂量膏方组大鼠腰椎骨密度、最大压缩载荷,血清PINP、OPG水平,骨组织Notch1、Notch2、Jagged1、Jagged2的mRNA和蛋白表达,OPG的mRNA表达显著升高(P<0.05),而血清TRACP、CTX-Ⅰ水平,骨组织RANKL、RANK、TRAF6的mRNA表达显著降低(P<0.05)。结论高剂量益肾健骨膏能改善去卵巢骨质疏松大鼠骨密度、骨强度及骨代谢,其作用机制可能与Notch通路及OPG/RANKL/RANK系统有关。

Objective To investigate the effect and mechanism of Yishenjiangu Cream Formula on bone metabolism in ovariectomized osteoporosis rats.Methods 50 female SD rats were randomly divided into sham operation group,model group,Yishenjiangu low-dose group,Yishenjiangu high-dose group and positive drug group.Rats in sham operation group and model group were gavaged with 10 mL/(kg·d)of saline;rats in Yishenjiangu low-dose and high-dose groups were administered with 2 g/(kg·d)and 13 g/(kg·d)Yishenjiangu Cream Formula respectively;rats in positive drug group were gavaged with 6.25 mg/(kg·w)alendronate D3 tablets.After 12 weeks of treatment,the bone mineral density(BMD)of lumbar spine and the maximum compressive load of lumbar spine were detected.tartrate-resistant acid phosphatase(TRACP),osteoprotegerin(OPG),type I procollagen amino-terminal peptide(PINP),type I collagen carboxy-terminal peptide(CTX-Ⅰ)in serum were tested by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expression of Notch1,Notch2,Jagged1 and Jagged2 were detected with Real-time PCR and Western blotting in bone tissue.The mRNA of OPG,RANK,RANKL and TRAF6 were detected with Real-time PCR in bone tissue.Results After 12-week treatment with high-dose Yishenjiangu Cream Formula,the BMD,maximum compressive load of lumbar spine,the serum levels of PINP and OPG,the mRNA and protein expression of Notch1,Notch2,Jagged1 and Jagged2,the relative mRNA levels of OPG in bone tissue were significantly increased(P<0.05),whereas the serum levels of TRACP and CTX-Ⅰ,the relative mRNA levels of RANKL,RANK,TRAF6 were were significantly decreased(P<0.05),compared with the model group.Conclusion High-dose Yishenjiangu Cream Formula can improve bone mineral density,bone strength and bone metabolism in ovariectomized osteoporosis rats,and its mechanism may be related to Notch pathway and OPG/RANKL/RANK system.