防己诺林碱对去卵巢小鼠骨代谢及骨微结构的影响

Effects of fangchinoline on bone metabolism and bone microarchitecture in ovariectomized mice

目的探讨防己诺林碱对去卵巢小鼠骨质疏松的保护作用。方法将40只C57BL/6小鼠随机分为空白(假手术)对照组、模型(去卵巢)组、阳性(雌二醇,E2)对照组、防己诺林碱组共4组,每组10只。连续腹腔给药7周后处死小鼠,取股骨及外周血清,通过ELISA法检测骨代谢相关血清学指标:抗酒石酸酸性磷酸酶(TRAcP)、I型胶原羧基末端肽(CTX)及I型胶原氨基末端肽(NTX);通过Micro-CT评估各组小鼠骨组织微结构相关指标:骨小梁百分比(BV/TV)、骨小梁数量(Tb.N)、骨小梁分离度(Tb.Sp)和骨小梁厚度(Tb.Th);通过实时荧光定量PCR检测小鼠骨质疏松骨吸收相关基因的表达情况,包括TRAcP、组织蛋白酶K(Cathepsin K)、活化T细胞核因子1(NFATc1)以及降钙素受体(CTR)。结果与去卵巢模型组比较,防己诺林碱治疗组小鼠BV/TV、Tb.N、Tb.Th显著升高,Tb.Sp明显降低,骨代谢相关指标(TRAcP、CTX、NTX)显著降低,破骨细胞标志基因(TRAcP、Cathepsin K、NFATc1以及CTR)表达水平明显下降。结论防己诺林碱对去卵巢小鼠骨质疏松有保护作用,有望为骨质疏松的临床用药提供新思路。

Objective To investigate the protective effect of fangchinoline on ovariectomy-induced osteoporosis.Methods Forty C57BL/6 mice were randomly divided into 4 groups,including blank control(sham)group,ovariectomized(OVX)mice model group,E2-treated group,and fangchinoline-treated group.Each group contained 10 mice.After 7 weeks of abdominal administration,all the mice were sacrificed.Serum was collected for detecting the following osteoporosis related biomarkers:tartrate-resistant acid phosphatase(TRAcP),C-terminal peptide of type I collagen(CTX),and N-terminal telopeptide of type I collagen(NTX).Femurs were removed for micro-CT analysis of the following bone mass index:bone volume/tissue volume(BV/TV),trabecular number(Tb.N),trabecular separation(Tb.Sp),and trabecular thickness(Tb.Th).Real-time PCR was applied to detect the expression of osteoporosis specific genes in femurs,including TRAcP,Cathepsin K,nuclear factor of activated T cells cytoplasmic 1(NFATc1),and calcitonin receptor(CTR).Results Compared with OVX mice model,fangchinoline-treated group had increased levels of BV/TV,Tb.N,and Tb.Th,and decreased level of Tb.Sp.Also,decreased levels of bone resorption related biomarkers,including TRAcP,CTX,and NTX,were found in fangchinoline-treated group.In addition,real-time PCR result showed that fangchinoline-treated group had decreased levels of osteoclast marker genes including TRAcP,Cathepsin K,NFATc1,and CTR.Conclusion Fangchinoline protects against osteoporosis in OVX mice and might provide new ideas for the clinical treatment of osteoporosis.