FZD2激活β-catenin稳定雌激素受体α缓解骨质疏松症的分子机制研究  

Molecular mechanism of FZD2 in alleviating osteoporosis by stabilizing estrogen receptor alpha via activatingβ-catenin

作　　者：崔永建[1] 李艳[2] 王桂芝[3] CUI Yongjian;LI Yan;WANG Guizhi(Department of General Medicine(Geriatric),the Sixth Affiliated Hospital of Xinjiang Medical University,Xinjiang,Urumqi 830002,China;不详)

机构地区：[1]新疆医科大学第六附属医院全科医学(老年病)科,乌鲁木齐市830002 [2]新疆医科大学第六附属医院急诊科,乌鲁木齐市830002 [3]新疆医科大学第六附属医院教学科,乌鲁木齐市830002

出　　处：《河北医药》2025年第2期201-206,共6页Hebei Medical Journal

基　　金：新疆维吾尔自治区自然科学基金资助项目(编号:2022D01C173)。

摘　　要：目的评估和探索FZD2增加双侧卵巢切除术(ovariectomy,OVX)小鼠模型骨量的功效以及其作用机制。方法建立OVX诱导的骨质疏松症小鼠模型。腺病毒过表达FZD2。30只6~8周龄雌性C57BL/6J小鼠随机分为Control组、Sham组、OVX Model组、OVX Model+E2 Treatment组[给予小鼠雌二醇(E2)处理]、OVX Model+FZD2 Treatment-L组(给予小鼠左后肢膝关节注射腺病毒-过表达-FZD2治疗)、OVX Model+control-R组(给予小鼠右后肢膝关节注射腺病毒-vector处理),每组6只(OVX Model+FZD2 Treatment-L组和OVX Model+Control-R组使用同样小鼠)。微计算机断层扫描测定小鼠左右后肢胫骨骨小梁和皮质骨骨量相关指标。Western blot测定小鼠胫骨FZD2、active-β-catenin、β-catenin、p-ERα、ERα、Runx2和ALP表达水平。免疫共沉淀法测定分别使用Wnt3a、E2和腺病毒-过表达-FZD2处理的小鼠胚胎成骨细胞MC3T3-E1中ERα和β-catenin的直接相互作用。结果与OVX Model组比较,OVX Model+E2 Treatment组小鼠骨组织中的FZD2、active-β-catenin、β-catenin、p-ERα、ERα、Runx2和ALP表达水平上调(P<0.05);骨小梁的骨体积分数和骨小梁数量数值升高(P<0.05);骨小梁分离和皮质骨的骨髓腔面积数值降低(P<0.05)。与OVX Model+E2 Treatment组比较,OVX Model+FZD2 Treatment-L组能够实现与OVX Model+E2 Treatment组类似的功效(P>0.05)。分别加入Wnt3a或E2或腺病毒-过表达-FZD2处理MC3T3-E1细胞后,co-IP免疫印迹结果显示ERα和β-catenin条带显影均为阳性。结论过表达和激活FZD2激活β-catenin稳定雌激素受体α缓解骨质疏松症。Objective To evaluate and explore the efficacy and mechanisms of Frizzled 2(FZD2)on increasing bone mass of mice with bilateral ovariectomy(OVX).Methods A mouse model of OVX-induced osteoporosis was established.Adenovirus overexpressing(OE)FZD2 was constructed.Thirty female C57BL/6J mice aged 6-8 weeks were randomly assigned into the Control group,Sham group,OVX Model group,OVX Model+E2 Treatment group(OVX+treatment of estradiol[E2]),OVX Model+FZD2 Treatment-L group(OVX+injection of adenovirus OE FZD2 into the mouse left hind knee joint)and OVX Model+Control-R group(OVX+injection of control vector into the mouse left hind knee joint),with 6 mice per group.Microcomputed tomography(Micro-CT)was used to measure the bone mass of tibial trabecular and cortical bone in the left and right hind limbs of mice.Expression levels of FZD2,active-β-catenin,β-catenin,phosphorylated-estrogen receptor alpha(p-ERα),ERα,Runt-related transcription factor 2(Runx2)and alkaline phosphatase(ALP)in the mouse tibia were determined by Western blot.The direct interaction of ERαandβ-catenin in mouse embryonic osteoblasts MC3T3-E1 treated with Wnt3a,E2 and adenovirus-OE-FZD2 was determined by co-immunoprecipination(co-IP).Results Compared with those of OVX Model group,FZD2,active-β-catenin,β-catenin,p-ERα,ERα,Runx2 and ALP in mouse bone tissue of OVX Model+E2 Treatment group were significantly up-regulated(P<0.05).Trabecular bone volume fraction(BV/TV%)and trabecular number(Tb.N)were significantly higher in the OVX Model+E2 Treatment group than those of OVX Model group(P<0.05).Trabecular separation(Tb.Sp)and cortical bone marrow area(Ma.Ar)were significantly lower in the OVX Model+E2 Treatment group than those of OVX Model group(P<0.05).Compared with OVX Model+E2 Treatment group,OVX Model+FZD2 Treatment-L group achieved similar efficacy as that in the OVX Model+E2 Treatment group(P>0.05).After the MC3T3-E1 cells were treated with Wnt3a or E2 or adenovirus-OE-FZD2,co-IP and Western blot results showed that ERαandβ-catenin were posi

关 键 词：骨质疏松症 雌激素 雌激素受体 Wnt/β-catenin信号 FZD2 

分 类 号：R579.1[医药卫生—消化系统]