巴戟天丸组方对Aβ损伤成骨细胞的作用及基于网络药理学的机制研究  

The roles of Bajitianwan formula on Aβ-injured osteoblasts and the mechanism based on network pharmacology

作　　者：姜涛 徐卫凡 蒋益萍 夏天爽 辛海量 JIANG Tao;XU Weifan;JIANG Yiping;XIA Tianshuang;XIN Hailiang(School of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Department of Pharmacognosy,School of Pharmacy,Naval Medical University,Shanghai 200433,China)

机构地区：[1]海军军医大学药学系生药学教研室,上海200433 [2]福建中医药大学药学院,福建福州350122

出　　处：《药学实践与服务》2024年第7期285-290,296,共7页Journal of Pharmaceutical Practice and Service

基　　金：国家自然科学基金(82004015,82174079);上海市卫健委中医药科研项目(2024QN020)。

摘　　要：目的 探讨巴戟天丸组方对Aβ损伤成骨细胞的骨形成作用及其机制。方法 以新生24 h Wistar大鼠所分离的成骨细胞为研究对象,用Aβ1-42寡聚体对成骨细胞进行损伤,并用巴戟天丸组方水提物进行药物干预。分别采用MTT法、碱性磷酸酶(ALP)活性检测、过氧化氢酶(CAT)活性检测、超氧化物歧化酶(SOD)活性检测、谷胱甘肽(GSH)活性检测以及丙二醛(MDA)活性检测。采用蛋白质印迹法检测骨形成相关蛋白骨形态发生蛋白2(BMP2)、成骨特异性转录因子(RUNX-2)、骨保护蛋白(OPG)的表达水平;明确巴戟天丸组方对Aβ损伤成骨细胞的作用后,采用网络药理学方法对潜在的作用机制进行预测。结果 巴戟天丸组方可显著促进Aβ损伤成骨细胞的增殖,提高ALP、SOD、GSH活性,抑制MDA活性,并促进骨形成相关蛋白BMP2、RUNX-2、OPG的表达。网络药理学分析显示,巴戟天丸组方发挥改善Aβ损伤成骨细胞的作用主要与AGE-RAGE信号通路、PI3K-Akt信号通路、MAPK信号通路以及神经活性配体与受体的相互作用通路等有关。结论 明确巴戟天丸组方具有改善Aβ损伤成骨细胞的作用,并通过网络药理学方法探究其相关作用通路,为传统方剂巴戟天丸抗骨质疏松的临床应用提供借鉴。Objective To explore the effect of Bajitianwan(BJTW)formula on bone formation of Aβ-injured osteoblasts and its mechanism.Methods Osteoblasts isolated from neonatal 24-hour Wistar rats were used for the study,and osteoblasts were subjected to damage with Aβ1-42 oligomers,and pharmacological intervention was performed with the aqueous extract of BJTW formula.The MTT assay,alkaline phosphatase(ALP)activity assay,catalase(CAT)activity assay,superoxide dismutase(SOD)activity assay,glutathione(GSH)activity assay and malondialdehyde(MDA)activity assay were carried out respectively.The expression levels of bone morphogenetic protein 2(BMP2),osteogenic specific transcription factor(RUNX-2)and osteoprotective protein(OPG)were detected by Western blotting.After confirming the effect of BJTW formula on Aβ-injured osteoblasts,the network pharmacology method was used to predict the potential pathways.Results The BJTW formula significantly promoted the proliferation of Aβ-injured osteoblasts,increased ALP,SOD and GSH activity,inhibited MDA activity,and promoted the expression of bone formation-related proteins BMP2,RUNX-2 and OPG.Network pharmacological analysis showed that the effect of ameliorating of Aβ-injured osteoblasts by BJTW formula was mainly mediated by AGE-RAGE,PI3K-Akt,MAPK and neuroactive ligand-receptor interaction signaling pathways.Conclusion In this study,the effect of BJTW formula on improving the osteoblasts damaged by Aβwas confirmed for the first time,and its related mechanism was explored based on network pharmacology method.The results lay a strong foundation for the clinical application of traditional formula BJTW against osteoporosis.

关 键 词：成骨细胞 巴戟天丸 AΒ沉积 网络药理学 

分 类 号：R285[医药卫生—中药学]