补肾健脾活血方对去卵巢大鼠BMP2/Smad信号通路的影响

Effect of Bushen Jianpi Huoxue Recipe on BMP2/Smad Signaling Pathway in Ovariectomized Rats

目的：研究补肾健脾活血方对去卵巢大鼠骨形态发生蛋白2（BMP2）/Smad信号通路的影响。方法：72只6月龄雌性SD大鼠,随机分为假手术组（24只）,手术组（48只）,双侧卵巢切除法造模,术后3个月两组各随机选取12只检测骨密度验证造模成功。手术组剩余的36只大鼠随机分为双侧卵巢切除模型组（OVX）,补肾健脾活血方组（BSJPHX,给药剂量2.979 g·kg^（-1））,阿仑膦酸钠维D3片（Ⅱ）组（ALN,给药剂量1.02 mg·kg^（-1））,假手术组,OVX组给与等体积生理盐水灌胃。12周后处死各组大鼠,双能X射线法检测大鼠全身骨密度,生物力学试验进行胫骨三点弯曲试验,实时荧光定量聚合酶链式反应（Real-time PCR）检测BMP2,Smad1,Runt相关转录因子2（Runx2）,骨保护素（OPG）基因的表达,蛋白免疫印迹法（Western blot）检测BMP2,p-Smad1,Runx2,OPG蛋白的表达。结果：造模3个月后,与假手术组比较,OVX组大鼠骨密度明显降低,最大载荷及刚度均降低,BMP2,Smad1,Runx2,OPG基因和蛋白表达水平显著降低（P〈0.05）;给药3个月后,与OVX组比较,BSJPHX组,ALN组骨密度均明显增高,最大载荷及刚度均增加,能显著提高BMP2,Smad1,Runx2,OPG基因和蛋白表达水平（P〈0.05）。结论：补肾健脾活血方既能通过调控BMP2/Smad通路的信号转导,又能上调OPG的表达,这可能是补肾健脾活血方防治绝经后骨质疏松症的机制之一。

Objective： To study the effect of Bushen Jianpi Huoxue recipe（ BSJPHX） on bone morphogenetic protein2（ BMP2）/Smad signaling pathway in ovariectomized rats. Method： Seventy-two female SD rats aged 6 months were randomly divided into sham group（ 24） and operation group（ 48）. The rats were ovariectomized to reproduce the model of osteoporosis. After 3 months,12 rats were randomly selected to measure bone mineral density,so as to verify successful modeling in each group. The remaining 36 rats in the operation group were randomly divided into ovarietomized（ OVX） group,BSJPHX（ 2. 979 g·kg^（-1）） group and alendronate sodium and vitamin D3 tablets（ ALN,1. 02 mg·kg^（-1）） group. After 12 weeks,bone mineral density in lumbar vertebrae was measured by dual energy X-ray. Biomechanical test was performed to tibia. Expressions of BMP2,Smad1,Runt related transcription factor 2（ Runx2）,and osteoprotegerin（ OPG） genes were detected by Realtime PCR. Expressions of BMP2,p-Smad1,Runx2,OPG protein was detected by Western blot. Result： After 3 months of modeling,compared with sham group,the BMD of rats in OVX group was significantly lower,both the maximum load and rigidity reduced（ P〈0. 05）; after 3 months of administration,compared with OVX group,the bone mineral density of BSJPHX and ALN groups was significantly higher,both the maximum load and rigidity increased,and expression levels of BMP2,Smad1,Runx2,and OPG genes significantly increased（ P〈0. 05）.Conclusion： BSJPHX can not only regulate BMP2/Smad pathway,but also up-regulate the expression of OPG,which may be one of the mechanisms of BSJPHX in prevention and treatment of postmenopausal osteoporosis.