紫草素介导Wnt/β-连环蛋白信号改善糖皮质激素性骨质疏松小鼠骨量  

Shikonin protects bone reduction in glucocorticoid-induced osteoporosis mice via activating Wnt/B-cateninsignaling

作　　者：宁永明[1] 王润政 严旭[1] 王永魁 崔妙然 毛克亚[2] 唐海[3] 孟斌[4] 刘宏建[1] Ning Yongming;Wang Runzheng;Yan Xu;Wang Yonghui;Cui Miaoran;Mao Keya;Tang Hai;Meng Bin;Liu Hongjian(Department of Orthopedics,the First Afiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Orthopedics,Chinese PLA General Hospital,Beijing 100853,China;Department of Orthopedics,Beijing Friendship Hospital of Capital Medical University,Bejing 100050,China;Department of Orthopedics,the First Afiliated Hospital of Soochow University,Suzhou 215006,China)

机构地区：[1]郑州大学第一附属医院骨科,郑州450052 [2]中国人民解放军总医院骨科,北京100853 [3]首都医科大学附属北京友谊医院骨科,北京100050 [4]苏州大学附属第一医院骨科,苏州215006

出　　处：《中华实验外科杂志》2023年第5期903-906,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81802128)。

摘　　要：目的探讨紫草素(SK)对糖皮质激素性骨质疏松症的潜在治疗作用和调控机制。方法按照1 mg/kg剂量给予雄性C57BL/6小鼠肌注地塞米松(DEX)诱导构建糖皮质激素性骨质疏松症(GIOP)模型,micro-CT检测骨微结构,免疫组织化学检测骨小梁osterix(OSX)和osteocalcin(OCN)表达;蛋白质印迹法(Western blot)检测骨组织中Wnt/β-连环蛋白(β-catenin)信号相关蛋白表达。培养小鼠骨髓间充质干细胞(BMSCs),用DEX(1 mol/L)、SK(50、100、200 mol/L)处理BMSCs;细胞计数试剂盒(CCK-8)检测细胞活性;茜素红染色法检测成骨矿化能力;用Wnt信号抑制剂XAV939评估紫草素通过Wnt/β-catenin信号通路治疗GIOP。两组间比较采用t检验,多组间比较采用单因素方差分析。结果Micro-CT结果显示,对照组、DEX组和DEX+SK组小鼠骨体积分数分别为(24.66±16.31)%、(14.76±3.99)%、(21.04±9.73)%,组间比较差异有统计学意义(F=10.03,P<0.05);皮质骨厚度分别为(0.60±0.01)、(0.30±0.01)、(0.52±0.01)mm,组间比较差异有统计学意义(F=11.13,P<0.05);骨小梁数量分别为(2.92±0.38)、(1.34±0.10)、(2.66±0.27)/mm,组间比较差异有统计学意义(F=12.54,P<0.05);骨小梁厚度分别为(0.210±0.002)、(0.100±0.001)、(0.190±0.001)mm,组间比较差异有统计学意义(F=13.48,P<0.05);骨小梁间隙分别为(0.280±0.003)、(0.530±0.003)、(0.340±0.004)mm,组间差异有统计学意义(F=21.80,P<0.05);免疫组织化学结果显示,与对照组比较、DEX组OSX和OCN表达下调;与DEX组比较,DEX+SK组OSX和OCN表达上调,差异有统计学意义(F=60.42,56.38,P<0.05)。Western blot结果显示DEX组小鼠Wnt1和磷酸化糖原合成激酶3β(p-GSK-3β)蛋白表达下降,而DEX+SK组小鼠Wnt1和p-GSK-3β蛋白表达上调。CCK-8结果显示,与对照组[(97.33±1.61)%]比较,DEX组细胞活性受到抑制[(47.30±3.98)%],与DEX组比较,DEX+SK(50 mol/L)组、DEX+SK(100 mol/L)组和DEX+SK(200 mol/L)组细胞活性上调,差异有统计学意义(t=36.87、5.16、3Objective To eexxpplolorree tthhee ppootteennttiiaall tthheerraappeeuuttiicc effects and regulatory mechanisms of shikonin(SK)on glucocorticoid-induced osteoporosis(GIOP).Methods Male C57BL/6 mice were administered intramuscular dexamethasone(DEX)at a dose of 1 mg/kg to induce GIOP model.Bone structure parameters were measured by micro-CT.Immunohistochemistry was used to detect the expression of osteogenic related proteins Osterix(OSX)and Osteocalcin(OCN)in bone trabeculae.Western blotting was applied to detect Wnt/β-catenin signaling related proteins.The mouse bone marrow mesenchymal stem cells(BMSCs)were cultured and treated with DEX(1 mol/L)and SK(50,100,200 mol/L).Cell counting kit-8(CCK-8)was used to detect cell activity.Alizarin red staining was used to detect osteogenic mineralization ability.The Wnt signaling inhibitor XAV939 was used to evaluate the curative effect of SK for GIOP via the Wnt/β-catenin signaling pathway.The comparison between the two groups was conducted using t-test,and the comparison between multiple groups was conducted using one-way ANOVA.Results Micro-CT results showed that BV/TV in control group,DEX group,and DEX+SK group was(24.66±16.31)%,(14.76±3.99)%,and(21.04±9.73)%,respectively,with statistically significant differences between the groups(F=10.03,P<0.05).The cortical bone thickness in control group,DEX group,and DEX+SK group was(0.60±0.01),(0.30±0.01),and(0.52±0.01)mm,respectively,with statistically significant differences between the groups(F=11.13,P<0.05).The number of bone trabeculae per 1 mm unit length in control group,DEX group,and DEX+SK group was 2.92±0.38,1.34±0.10,and 2.66±0.27,respectively,with statistically significant differences between groups(F=12.54,P<0.05).The thickness of bone trabeculae in control group,DEX group,and DEX+SK group was(0.210±0.002),(0.100±0.001),and(0.190±0.001)mm,respectively,with statistically significant differences between groups(F=13.48,P<0.05).The bone trabecular spaces in control group,DEX group,and DEX+SK group were 0.280�

关 键 词：紫草素 糖皮质激素性骨质疏松 Wnt/β-连环蛋白 

分 类 号：R285.5[医药卫生—中药学]