Herb couplet medicines (Erxian) protect osteoblasts from high glucose-induced damage by reducing cell apoptosis in diabetic osteoporosis: A network pharmacology and experimental verification-based study

IF 1.9 4区 医学 Q3 INTEGRATIVE & COMPLEMENTARY MEDICINE
Xuan Zhou , Mengying Li , Minjie Zhang , Yang Zhang
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Abstract

Introduction

Traditional Chinese medicine (TCM) has unique advantages in treating diabetic osteoporosis (DOP). This study aimed to explore the mechanisms underlying the protective effects of herb couplet medicines (Erxian) on DOP using network pharmacology and verify these mechanisms using cell experiments.

Methods

We used chromatography to analyze the bioactive components of serum containing Erxian couplet medicines (ECMs) and searched for possible targets of the active ingredients in the Traditional Chinese Medicine Systems Pharmacology database. The target proteins of DOP were retrieved from the GeneCards and Online Mendelian Inheritance in Man databases. A component-target network diagram was then constructed using the screened drug components and target information for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. In vitro, cell morphological changes were observed using alkaline phosphatase staining, and the effect of the ECMs on osteoblast viability was assessed using a cell counting kit-8 assay. Finally, the pathway information predicted by network pharmacology was validated using flow cytometry, real-time polymerase chain reaction, and western blot experiments.

Results

Twelve types of active ingredients in serum containing ECMs were obtained via chromatography, and 96 potential targets were identified using network pharmacology analysis. Furthermore, GO and KEGG pathway enrichment analyses revealed that the ECMs might participate in the apoptosis signaling pathway in the treatment of DOP. In vitro tests showed that treatment with ECM serum increased osteoblasts’ survival rate and improved the HG group's cell morphology (P < 0.05). BCL2-associated X protein (BAX) expression increased in the HG group but decreased after ECM serum addition (P < 0.05). The expression of B-cell lymphoma-2 (BCL-2) was found to be reduced in the HG group but increased after ECM serum addition (P < 0.05).

Conclusion

ECMs can treat DOP via multiple targets and multiple signaling pathways. Moreover, the mechanism underlying the effects of ECMs on osteoblasts was verified by inhibiting apoptotic. The complete signaling pathway requires future study.

Abstract Image

草本对联药物(二仙)通过减少糖尿病骨质疏松症患者的细胞凋亡,保护成骨细胞免受高糖诱导的损伤:基于网络药理学和实验验证的研究
引言中医药在治疗糖尿病骨质疏松症(DOP)方面具有独特的优势。方法我们用色谱法分析了含有二仙对联药物(ECMs)的血清中的生物活性成分,并在中药系统药理学数据库中搜索了有效成分的可能靶点。从 GeneCards 和 Online Mendelian Inheritance in Man 数据库中检索了 DOP 的靶蛋白。然后利用筛选出的药物成分和靶标信息构建成分-靶标网络图,进行基因本体(GO)和京都基因组百科全书(KEGG)富集分析。在体外,使用碱性磷酸酶染色观察细胞形态变化,并使用细胞计数试剂盒-8 评估 ECM 对成骨细胞活力的影响。最后,利用流式细胞术、实时聚合酶链反应和 Western 印迹实验验证了网络药理学预测的通路信息。结果通过色谱法获得了含有 ECMs 的血清中的 12 种活性成分,并利用网络药理学分析确定了 96 个潜在靶点。此外,GO 和 KEGG 通路富集分析表明,ECMs 可能参与了治疗 DOP 的细胞凋亡信号通路。体外试验显示,用 ECM 血清治疗可提高成骨细胞的存活率,并改善 HG 组细胞的形态(P < 0.05)。BCL2- 相关 X 蛋白(BAX)的表达在 HG 组中增加,但在添加 ECM 血清后则减少(P < 0.05)。结论ECMs 可通过多靶点和多信号通路治疗 DOP。此外,ECMs 通过抑制成骨细胞凋亡验证了其作用机制。完整的信号通路还需要进一步研究。
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来源期刊
European Journal of Integrative Medicine
European Journal of Integrative Medicine INTEGRATIVE & COMPLEMENTARY MEDICINE-
CiteScore
4.70
自引率
4.00%
发文量
102
审稿时长
33 days
期刊介绍: The European Journal of Integrative Medicine (EuJIM) considers manuscripts from a wide range of complementary and integrative health care disciplines, with a particular focus on whole systems approaches, public health, self management and traditional medical systems. The journal strives to connect conventional medicine and evidence based complementary medicine. We encourage submissions reporting research with relevance for integrative clinical practice and interprofessional education. EuJIM aims to be of interest to both conventional and integrative audiences, including healthcare practitioners, researchers, health care organisations, educationalists, and all those who seek objective and critical information on integrative medicine. To achieve this aim EuJIM provides an innovative international and interdisciplinary platform linking researchers and clinicians. The journal focuses primarily on original research articles including systematic reviews, randomized controlled trials, other clinical studies, qualitative, observational and epidemiological studies. In addition we welcome short reviews, opinion articles and contributions relating to health services and policy, health economics and psychology.
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