人参皂苷对模拟失重巨噬细胞的保护作用研究

Protective effects of ginsenosides on macrophages subjected to simulated weightlessness

目的探究失重条件下炎症的作用机制及人参皂苷对巨噬细胞的炎症保护作用。方法首先通过GeneCards数据库检索与失重、炎症和免疫相关的基因,再通过基因/蛋白相互作用检索搜查工具(STRING)蛋白网络分析,寻找失重诱导炎症的核心靶点。通过非标记定量蛋白组学(LFQ)寻找人参皂苷处理人单核细胞白血病(THP-1)细胞的差异基因,通过基因本体(GO)、京都基因与基因组百科全书(KEGG)富集分析研究人参皂苷主要参与的生物过程和信号通路及人参皂苷针对失重性炎症的主要作用靶点。最后采用酶联免疫吸附试验(ELISA)法检测人参皂苷对脂多糖(LPS)诱导的THP-1细胞模拟失重期及失重恢复期白细胞介素(IL)-1β、IL-6、IL-8和肿瘤坏死因子α(TNF-α)的分泌影响。结果通过GeneCards数据库检索到炎症相关基因2933个,失重相关基因425个,免疫相关基因4564个。通过蛋白相互作用网络(PPI)确定了与失重诱导的炎症相关的靶点,如IL-1β、IL-6、TNF-α和白蛋白(ALB)。通过蛋白组学研究发现人参皂苷主要参与多种炎症相关信号通路及病原微生物感染通路,并显著改变CD40、IL-1β和腺苷二磷酸核糖聚合酶(PARP1)等蛋白表达。细胞模拟失重试验显示人参皂苷可以显著降低模拟失重期LPS诱导单核/巨噬细胞THP-1炎性因子IL-6、TNF-α的分泌(P<0.0001),同时还可降低失重恢复期IL-1β、IL-6、IL-8和TNF-α的分泌(P<0.0001)。结论失重可影响多种炎症相关信号通路,而人参皂苷可减少模拟失重巨噬细胞多种炎性因子的分泌起到缓解炎症的作用。本研究为进一步探究人参皂苷应用于防护失重条件下LPS诱导的炎症提供理论基础。

Objective To investigate the evolution of inflammation under conditions and the effects of ginsenosides on macrophages subjected to the simulated weightlessness,with the aim of mitigating the inflammation.Methods Initially,genes related to weightlessness,inflammation,and immunity were identified in the GeneCards database.Then,Search Tool for the Retrieval of Interaction Gene/Proteins(STRING)protein network analysis was conducted to determine the core targets involved in the weightlessness-induced inflammation.Subsequently,Label-Free Quantitative(LFQ)proteomics was carried out to discern the distinctive genes within ginsenoside-treated Tohoku Hospital Pediatrics-1(THP-1)cells.Next,utilizing the outcomes of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,the biological processes and signaling pathways in which ginsenosides predominately engaged were scrutinized,and the primary targets of ginsenosides in combating weightlessness-induced inflammation were examined.Finally,enzyme-linked immunosorbent assay(ELISA)was performed to detect the secretion levels of interleukin(IL)-1β,IL-6,IL-8,and tumor necrosis factor(TNF)-αfrom lipopolysaccharide(LPS)-induced THP-1 cells under simulated weightlessness conditions,as well as during the weightlessness recovery period following treatment with ginsenosides.Results A total of 2933 genes associated with inflammation,425 genes linked to weightlessness,and 4564 genes connected to immunity were retrieved from the GeneCards database.Protein-protein interaction(PPI)networks were generated to identify pivotal targets associated with weightlessness-induced inflammation such as IL-1β,IL-6,TNF,and albumin(ALB).It was found that ginsenosides primarily participated in the regulation of various inflammationrelated signaling pathways and pathways related to pathogenic microorganism infections.Moreover,it has a significant impact on the expression of proteins such as cluster of differentiation 40(CD40),IL-1β,and poly ADP-ribose polymerase 1(PARP1).As revealed in the simulated weightlessness cell test,ginsenosides exhibited a remarkable capacity to attenuate the secretion of inflammatory factors,specifically IL-6 and TNF-α(P<0.0001),in THP-1 macrophages following induction by LPS under simulated weightlessness conditions.In addition,it reduced the secretion of IL-1β,IL-6,IL-8,and TNF-α(P<0.0001)during the weightlessness recovery phase.Conclusion Weightlessness can disrupt several inflammation-related signaling pathways,but ginsenosides were shown to mitigate the release of various inflammatory factors in macrophages subjected to simulated weightlessness,thereby exerting a protective role against inflammation.This study has laid a theoretical groundwork for further exploring the potential application of ginsenosides in safeguarding against LPS induced inflammation in a weightlessness environment.