基于Wnt/β-catenin信号通路介导的自噬和凋亡探讨苦豆子碱对骨质疏松小鼠骨代谢的影响  

To investigate the effect of aloperine on bone metabolism in osteoporotic mice based on autophagy and apop⁃tosis mediated by Wnt/β⁃catenin signaling pathway

作　　者：肖知周 黄莺 卞华伟[1] XIAO Zhizhou;HUANG Ying;BIAN Huawei(Department of Clinical Nutrition,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,Guangdong,China;不详)

机构地区：[1]中山大学附属第三医院临床营养科,广东广州510630 [2]广州医科大学附属第一医院营养科,广东广州510120

出　　处：《实用医学杂志》2025年第4期500-508,共9页The Journal of Practical Medicine

基　　金：广东省重点领域研发计划项目(编号:2019B020213001)。

摘　　要：目的本研究旨在探讨苦豆子碱(aloperine,ALO)通过Wnt/β-catenin信号通路介导的自噬和凋亡过程对骨质疏松(osteoporosis,OP)小鼠骨代谢的影响。方法将60只小鼠随机分成6组,每组10只:Sham组(假手术小鼠)、OP组(切除双侧卵巢建立骨质疏松症模型)、L-ALO组(OP小鼠腹腔注射10 mg/kg苦豆子碱)、M-ALO组(OP小鼠腹腔注射20 mg/kg苦豆子碱)、H-ALO组(OP小鼠腹腔注射30 mg/kg苦豆子碱)和EV组(OP小鼠灌胃0.09 mg/kg戊酸雌二醇),造模给药。检测小鼠胫骨的骨密度(BMD)与骨微结构。HE染色观察胫骨骨组织形态。ELISA检测小鼠血清中OCN、OPG、ALP、Ca和P的水平变化。免疫蛋白印迹检测LC3-Ⅱ/LC3-Ⅰ、Beclin-1、P62、Caspase-3、Caspase-9、Bax、Wnt3a、β-catenin和C-Myc蛋白表达水平。免疫荧光检测自噬小体。结果与Sham组相比,OP组的BMD和骨小梁厚度显著降低,骨小梁分离度和骨表面积与体积显著增加(P<0.05);OPG、OCN、Ca和P水平显著下调,ALP水平显著上调(P<0.05);LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1、Wnt3a、β-catenin和C-Myc蛋白表达水平显著降低,P62、Caspase-3、Caspase-9和Bax蛋白表达水平显著增加(P<0.05)。与OP组相比,L-ALO组、M-ALO组和H-ALO组的BMD和骨小梁厚度显著增加,骨小梁分离度和骨表面积与体积显著降低(P<0.05);OPG、OCN、Ca和P水平显著上调,ALP水平显著下调(P<0.05);LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1、Wnt3a、β-catenin和C-Myc蛋白表达水平显著增加,P62、Caspase-3、Caspase-9和Bax蛋白表达水平显著降低(P<0.05),并呈剂量依赖性。与OP组相比,EV组的BMD和骨小梁厚度显著增加,骨小梁分离度和骨表面积与体积比显著降低(P<0.05);OPG、OCN、Ca和P水平显著上调,ALP水平显著下调(P<0.05);LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1、Wnt3a、β-catenin和C-Myc蛋白表达水平显著增加,P62、Caspase-3、Caspase-9和Bax蛋白表达水平显著降低(P<0.05)。结论苦豆子碱可能通过促进成骨细胞自噬,并抑制其凋亡来Objective To investigate the effects of aloperine(ALO)on bone metabolism in osteoporosis(OP)mice via Wnt/β⁃catenin signaling pathway⁃mediated autophagy and apoptosis.Methods Sixty mice were randomly allocated into six groups(n=10 per group):Sham group(sham⁃operated mice),OP group(osteoporosis model induced by bilateral ovariectomy),L⁃ALO group(OP mice intraperitoneally injected with 10 mg/kg aloperine),M⁃ALO group(OP mice intraperitoneally injected with 20 mg/kg aloperine),H⁃ALO group(OP mice intraperitoneally injected with 30 mg/kg aloperine),and EV group(OP mice administered 0.09 mg/kg estradiol valerate).Bone mineral density and microstructure of the tibia were assessed.Hematoxylin and eosin(HE)staining was per⁃formed to examine the morphology of tibial bone tissue.Serum levels of OCN,OPG,ALP,Ca,and P were measured using ELISA.Protein expression levels of LC3-Ⅱ,LC3-Ⅰ,Beclin⁃1,P62,Caspase⁃3,Caspase⁃9,Bax,Wnt3a,β⁃catenin,and C⁃Myc were analyzed by Western blot.Autophagosomes were visualized using immunofluorescence.Results Compared with the Sham group,the bone mineral density(BMD)and trabecular thickness in the OP group were significantly reduced,while trabecular separation,bone surface area,and volume were significantly increased(P<0.05).The levels of OPG,OCN,Ca,and P were significantly downregulated,whereas ALP levels were significantly upregulated(P<0.05).Additionally,the LC3⁃Ⅱ/LC3⁃Ⅰratio and expression levels of Beclin⁃1,Wnt3a,β⁃catenin,and C⁃Myc proteins were significantly decreased,while the expression levels of P62,Caspase⁃3,Caspase⁃9,and Bax proteins were significantly increased(P<0.05).Compared with the OP group,the L⁃ALO,M⁃ALO,and H⁃ALO groups exhibited significant increases in BMD and trabecular thickness,along with significant decreases in trabecular separation,bone surface area,and volume(P<0.05).The levels of OPG,OCN,Ca,and P were significantly upregulated,while ALP levels were significantly downregulated(P<0.05).Furthermore,the LC3⁃Ⅱ/LC3�

关 键 词：苦豆子碱 骨质疏松 Wnt/β-catenin信号通路 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]