基于MAPK/NF-κB信号通路探究石杉碱甲对去卵巢小鼠骨质疏松的作用  

Effect and mechanism of huperzine A on osteoporosis in ovariectomized mice based on MAPK/NF⁃κB signaling pathway

作　　者：郑锦畅 林思恩 梁振 魏波 胡资兵 林瀚 孙杰聪 郭伟雄 陈驹 谢斌 黄成硕 ZHENG Jinchang;LIN Sien;LIANG Zhen;WEI Bo;HU Zibing;LIN Han;SUN Jiecong;GUO Weixiong;CHEN Ju;XIE Bin;HUANG Chengshuo(Orthopaedic Center,Affiliated Hospital of Guangdong Medical University,Zhanjiang 524001,China;Department of Pharmacy,Affiliated Hospital of Guangdong Medical University)

机构地区：[1]广东医科大学附属医院骨科中心,湛江524001 [2]广东医科大学附属医院药学部

出　　处：《山西医科大学学报》2025年第1期49-56,共8页Journal of Shanxi Medical University

基　　金：广东省医学科学技术研究基金项目(B2024110,B2024006);湛江市科技发展专项资金竞争性分配项目(2022A01176)。

摘　　要：目的 探讨石杉碱甲对去卵巢小鼠骨质疏松的作用及其机制。方法 40只雌性C57BL/6小鼠随机分为假手术组(sham)、去卵巢组(OVX)、石杉碱甲低剂量组(SSJJ-L,0.05 mg/kg)和石杉碱甲高剂量组(SSJJ-H,0.2 mg/kg)。采用双侧卵巢切除法构建骨质疏松模型。造模2个月后取材,ELISA法检测血清肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白介素1β(interleukin 1β,IL-1β)、IL-6和IL-10水平,化学发光法检测血清碱性磷酸酶(alkaline phosphatase,ALP)和抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase, StrACP)水平,镜下观察胫骨上段骨小梁面积百分比和骨形成动态变化,压缩法测定股骨的生物力学,以及蛋白质免疫印迹法(Western blot,WB)检测MAPK/NF-κB通路关键蛋白表达水平。结果 与sham组比较,OVX组小鼠血清TNF-α、IL-1β、IL-6、IL-10、StrACP和ALP水平明显升高(P<0.01),胫骨上段骨小梁面积百分数明显降低(P<0.01),双荧光间距明显增加(P<0.01),股骨最大载荷和断裂载荷明显降低(P<0.01),股骨p-p38 MAPK和p-NF-κB p65蛋白相对表达水平明显升高(P<0.01)。与OVX组相比,石杉碱甲给药组小鼠血清TNF-α、IL-1β、IL-6、IL-10、StrACP和ALP水平明显降低(P<0.01),胫骨上段骨小梁面积百分数明显升高(P<0.01),双荧光间距明显降低(P<0.01),股骨最大载荷和断裂载荷明显升高(P<0.01),股骨p-p38 MAPK和p-NF-κB p65蛋白相对表达水平明显降低(P<0.01)。与石杉碱甲低剂量组相比,石杉碱甲高剂量组小鼠上述指标均明显改善(P<0.05)。结论 石杉碱甲可能通过调控MAPK/NF-κB信号通路减轻炎症,抑制骨吸收和促进骨形成,改善骨质疏松。Objective To investigate the effect and mechanism of huperzine A on osteoporosis in ovariectomized mice.Methods Forty female C57BL/6 mice were randomly divided into sham group,OVX group,huperzine A low-dose group(SSJJ-L,0.05 mg/kg)and huperzine A high-dose group(SSJJ-H,0.2 mg/kg).The osteoporosis model was established by bilateral ovariectomy.After two months of modeling,the levels of tumor necrosis factor-α(TNF-α),interleukin 1β(IL-1β),IL-6,and IL-10 in serum were detected by ELISA.Alkaline phosphatase(ALP)and tartrate-resistant acid phosphatase(StrACP)levels in serum were detected by chemilumines-cence assay.Dynamic changes of bone trabecular area and bone formation in upper tibia were observed under the microscope.The biome-chanics of femur were measured by compression method and the expression level of key proteins in MAPK/NF-κB pathway was detected by Western blot.Results Compared with sham group,the serum levels of TNF-α,IL-1β,IL-6,IL-10,StrACP and ALP were sig-nificantly increased in OVX group(P<0.01),the percentage of trabecular bone area in upper tibia was significantly decreased(P<0.01),the double fluorescence distance was significantly increased(P<0.01),the maximum load and the fracture load of femur were significantly decreased(P<0.01),and the relative expression levels of p-p38 MAPK and p-NF-κB p65 were significantly increased(P<0.01).Compared with OVX group,the serum levels of TNF-α,IL-1β,IL-6,IL-10,StrACP and ALP were significantly decreased in huperzine A group(P<0.01),the percentage of bone trabecular area in upper tibia was significantly increased(P<0.01),the double fluorescence distance was significantly decreased(P<0.01),the maximum load and the fracture load of femur were significantly in-creased(P<0.01),and the relative expression levels of p-p38 MAPK and p-NF-κB p65 protein were significantly decreased(P<0.01).Compared with huperzine A low-dose group,the above indexes in huperzine A high-dose group were significantly improved(P<0.05).Conclusion Huperzine A may regulate MAPK/NF-κ

关 键 词：石杉碱甲 骨质疏松 小鼠 卵巢摘除 MAPK/NF-κB信号通路 炎症 

分 类 号：R68[医药卫生—骨科学]