金天格防治肌少-骨质疏松症的作用机制

The mechanism of Jintiange in the prevention and treatment of osteosarcopenia

目的 基于Akt/NF-κB/Bak信号通路,探讨金天格(JTG)防治肌少-骨质疏松症(OS)的作用机制。方法 药效团模型部分,采用高效液相色谱法(HPLC)测定金天格的成分,并对所得成分进行药效团模型分析。动物实验部分,将60只6月龄雌性SPF级SD大鼠随机分为正常组、假手术组、OS组、OS+金天格组、OS+雌二醇组。假手术组或模型组分别进行假手术或去势。术后1周,模型组大鼠腹腔注射地塞米松,连续2周。造模3个月后,药物干预组大鼠分别灌胃金天格或雌二醇水溶液,其他组灌胃等体积生理盐水。灌胃3个月后,各组大鼠分别进行前肢抓力检测、体成分检测、全身和局部骨密度检测、股骨远端骨微结构检测、Akt/NF-κB/Bak信号通路相关蛋白检测等。结果 高效液相色谱法鉴定出金天格含17种氨基酸。药效团模型发现这17种氨基酸可能通过调控PI3K-Akt和凋亡信号通路等防治OS。动物实验研究发现金天格明显改善OS大鼠前肢抓力、全身骨骼肌质量指数、全身及局部骨密度、股骨远端骨微结构等表型特征,增加OS大鼠腓肠肌p-Akt和NF-κB的蛋白表达,抑制促凋亡蛋白Bak和肌肉萎缩相关蛋白Fbx32蛋白表达。结论 金天格具有防治肌少-骨质疏松症的作用,其机制可能与激活Akt/NF-κB/Bak信号通路有关。

Objective To explore the mechanism of Jintiange(JTG) in preventing and treating osteosarcopenia(OS) based on the Akt/NF-κB/Bak signaling pathway. Methods In the pharmacophore model part, high performance liquid chromatography(HPLC) was used to determine the components of JTG. The pharmacophore model analysis of the obtained components was carried out. In the part of animal experiment, 60 6-month-old female SPF SD rats were randomly divided into Control group, Sham operation group, OS group, OS+JTG group, and OS+Egroup. Rats in the Sham group or the model group were performed Sham or OVX, respectively. One week after the operation, the rats in the model group were intraperitoneally injected with dexamethasone for two consecutive weeks. After 3 months of modeling, rats in the drug intervention group were intragastrically administered with JTG or Eaqueous solution. Rats in the other groups were intragastrically administered with equal volume of normal saline. After gavage for 3 months, rats in each group were tested for forelimb grip, body composition, whole body and local bone mineral density, distal femur bone microstructure, proteins related to Akt/NF-κB/Bak signaling pathway, etc. Results Seventeen kinds of amino acids in JTG were identified using HPLC. The pharmacophore model found that these 17 amino acids might prevent and treat OS by regulating PI3 K-Akt and apoptosis signaling pathways. Animal experiments found that JTG significantly improved the phenotypic characteristics of OS rats’ forelimb grip, whole body skeletal muscle mass index, whole body and local bone mineral density, and bone microstructure of the distal femur. JTG significantly increased the protein expression of p-Akt and NF-κB in gastrocnemius of OS rats, and inhibited the protein expression of pro-apoptotic protein Bak and muscle atrophy-related protein Fbx32. Conclusion JTG has the effect of preventing and treating OS, and its mechanism may be related to the activation of Akt/NF-κB/Bak signaling pathway.