龟甲水提液通过神经肽调控去卵巢大鼠骨形成的机制

Mechanism of Plastrum testudinis extracts regulating bone formation through neuropeptide in ovariectomized rat

目的:探讨龟甲水提液(PTE)是否通过影响骨、脑组织中神经肽Y(NPY)及其受体的表达从而促进去卵巢(OVX)大鼠骨形成。方法:30只雌性SD大鼠随机分为假手术组(Sham组)、OVX组和PTE组(80mg·kg^(-1)·d^(-1)),利用双侧OVx法建立骨质疏松模型,连续灌胃给药12周,收集各组大鼠股骨、全脑及下丘脑。采用Micro-CT和HE染色法观察股骨骨微结构变化,利用Westernblot和RT-PCR法检测股骨成骨分化相关因子,以及全脑、下丘脑和股骨NPY及其Y-Y1、Y-Y2受体(NPYIR、NPY2R)的表达水平。结果:PTE能显著改善OVX大鼠股骨骨微结构,促进OVX大鼠股骨组织中成骨分化相关因子ALP、BMP-2mRNA及蛋白,COL1、Runx2mRNA的表达(P<0.01,P<0.05),下调OVX大鼠全脑与下丘脑组织中NPYIRmRNA及蛋白表达(P<0.01),下调OVX大鼠全脑与股骨组织中NPYmRNA及蛋白表达(P<0.01,P<0.05),上调OVX大鼠股骨组织中NPY1R和NPY2R蛋白表达(P<0.01)。结论:PTE可能通过调控脑、下丘脑及股骨中的神经肽信号促进去卵巢大鼠骨形成。

Objective:To investigate whether Plastrum testudinis extracts(PTE)promote bone formation in the ovariectomized rat by affecting the expression of neuropeptide Y(NPY)and its receptors in bone and brain tissues.Methods:Thirty female SD rats were randomly divided into Sham group,ovariectomy group(OVX group)and PTE group(80 mg·kg^(-1)·d^(-1)).Bilateral ovariectomy was used to establish osteoporosis model,and continuous intragastric administration was performed in each group.After 12 weeks,the femur,whole brain and hypothalamus of rats were collected.Micro-CT and HE staining were used to observe the changes in microstructure of the femur.Western blot and RT-PCR were used to detect the expression of osteogenic related factors,as well as NPY and its Y-Y1 receptor(NPY1R),Y-Y2 receptor(NPY2R)in the whole brain,hypothalamus and femur.Results:PTE could significantly improved the microstructure of the femur in OVX rats,promoted the mRNA and protein expressions of ALP,BMP-2,the mRNA expression of COL1 and Runx2(P<0.01,P<0.05).Furthermore,the mRNA and protein expressions of neuropeptide NPYIR in the whole brain and hypothalamus,together with the expression of NPY protein in the whole brain and femur,were significantly decreased in OVX rats after treated with PTE(P<0.01,P<0.05).However,NPY1R and neuropeptide NPY2R protein were significantly increased in OVX rats after treated with PTE(P<0.01).Conclusion:PTE may promote bone formation in ovariectomized rats by regulating neuropeptide signals in the brain,hypothalamus and femur.