目的:本研究基于“肝与大肠相通”理论,采用网络药理学预测与动物实验验证相结合的策略,探讨赶黄草–炙黄芪–生大黄复方(SHY)干预酒精性肝损伤(ALI)的作用机制。方法:网络药理学获取SHY干预ALI的活性成分及主要靶点,构建PPI网络,进行GO...目的:本研究基于“肝与大肠相通”理论,采用网络药理学预测与动物实验验证相结合的策略,探讨赶黄草–炙黄芪–生大黄复方(SHY)干预酒精性肝损伤(ALI)的作用机制。方法:网络药理学获取SHY干预ALI的活性成分及主要靶点,构建PPI网络,进行GO和KEGG富集分析筛选可能涉及的生物过程和信号通路。利用Lieber-DeCarli酒精液体饲料[5% (vol/vol)乙醇]复制野生型(WT)小鼠ALI模型,通过检测体循环血清ALT、IL-11及肝脏组织IL-11和p-STAT3蛋白表达,验证SHY对ALI的保护作用及机制。结果:通过TCMSP数据库筛选共获得15种活性成分及219个药物靶点,与ALI疾病靶点交集得到106个关键靶点。PPI网络分析显示TNF、IL6、AKT1等为核心节点,GO/KEGG富集提示其通过IL-17、TNF等信号通路调控氧化应激与炎症反应。动物实验证实SHY显著降低模型小鼠血清ALT、IL-11水平(P Objective: Based on the traditional Chinese medicine theory of “liver-intestine correlation,” this study integrated network pharmacology prediction with experimental validation to investigate the mechanism of the Penthorum chinense Pursh-Astragalus membranaceus-Raw Rheum palmatum compound (SHY) in alleviating alcohol-induced liver injury (ALI). Methods: Network pharmacology was employed to identify active components and key targets of SHY against ALI. Protein-protein interaction (PPI) networks were constructed, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to elucidate biological processes and signaling pathways. ALI was induced in wild-type (WT) mice using the Lieber-DeCarli ethanol liquid diet (5% ethanol, vol/vol). Serum alanine aminotransferase (ALT) and interleukin-11 (IL-11) levels, along with hepatic IL-11 and phosphorylated STAT3 (p-STAT3) protein expression, were measured to validate SHY’s hepatoprotective effects. Results: Fifteen bioactive components and 219 potential targets of SHY were identified from the TCMSP database, with 106 overlapping targets linked to ALI. PPI network analysis revealed TNF, IL6, and AKT1 as core targets. GO/KEGG enrichment highlighted SHY’s regulation of oxidative stress and inflammation via IL-17 and TNF signaling pathways. Animal experiments demonstrated that SHY significantly reduced serum ALT and IL-11 levels (P < 0.01) and suppressed hepatic p-STAT3 phosphorylation. Western blot confirmed a 1.8-fold downregulation of IL-11 expression in SHY-treated mice. Conclusion: This study is the first to demonstrate that SHY ameliorates ALI by modulating the IL-11R/JAK/STAT axis, providing molecular evidence for the “treating liver diseases through intestinal regulation” theory.展开更多
AIM： To explore the effect of recombinant human interleukin-11 （rhIL-11） on the expressions of interleukin-11 receptor α-chain （IL-11Rα） and an additional signal transducer glycoprotein 130 （gp130） in intesti...AIM： To explore the effect of recombinant human interleukin-11 （rhIL-11） on the expressions of interleukin-11 receptor α-chain （IL-11Rα） and an additional signal transducer glycoprotein 130 （gp130） in intestinal epithelium cell line-6 （IEC-6） after neutron irradiation. METHODS： Cultured IEC-6 cells were exposed to 4.0Gy neutron and treated with 100 ng/mL rhIL-11 12 h prior to or immediately after irradiation. The apoptosis and necrosis rates and expressions of IL-11Rα and gp130 were observed by flow cytometry, immunohistochemistry, Western blot and image analysis. RESULTS： The apoptosis rate of IEC-6 cells was increased by irradiation at 6 h （P 〈 0.01）, IL-11 stimulation resulted in a decreased apoptosis rate in irradiated IEC-6 cells （P 〈 0.05）. In normal control IEC-6 cells, intense immunoreactivity of IL-11Rα was located within the cell membrane and cytoplasm. The level of IL-11Rα expression significantly decreased at 6 h after irradiation （P 〈 0.01） and restored at 24 h after irradiation. In IEC-6 cells treated with both radiation and rhIL-11, the level of IL- 11Rα expression was higher than that of irradiated cells （P 〈 0.05）. When it came to gp130 protein, it was located in the cytoplasm of IEC-6 cells. After irradiation, we found a progressive decrease in the expression of gp130 protein （P 〈 0.05） in 48 hours post-radiation, while in rhIL-11-stimulated cells, it came back to normal level at 24 h after irradiation and decreased at 48 h, but was still higher than that of only irradiated cells （P 〈 0.05）. CONCLUSION： rhIL-11 can protect IEC-6 cells from neutron irradiation. The protective effect of rhIL-11 might be connected with its ability to up-regulate the expressions of specific ligand-binding subunit IL-11Rα and signal-transducing subunit gp130.展开更多
文摘目的:本研究基于“肝与大肠相通”理论,采用网络药理学预测与动物实验验证相结合的策略,探讨赶黄草–炙黄芪–生大黄复方(SHY)干预酒精性肝损伤(ALI)的作用机制。方法:网络药理学获取SHY干预ALI的活性成分及主要靶点,构建PPI网络,进行GO和KEGG富集分析筛选可能涉及的生物过程和信号通路。利用Lieber-DeCarli酒精液体饲料[5% (vol/vol)乙醇]复制野生型(WT)小鼠ALI模型,通过检测体循环血清ALT、IL-11及肝脏组织IL-11和p-STAT3蛋白表达,验证SHY对ALI的保护作用及机制。结果:通过TCMSP数据库筛选共获得15种活性成分及219个药物靶点,与ALI疾病靶点交集得到106个关键靶点。PPI网络分析显示TNF、IL6、AKT1等为核心节点,GO/KEGG富集提示其通过IL-17、TNF等信号通路调控氧化应激与炎症反应。动物实验证实SHY显著降低模型小鼠血清ALT、IL-11水平(P Objective: Based on the traditional Chinese medicine theory of “liver-intestine correlation,” this study integrated network pharmacology prediction with experimental validation to investigate the mechanism of the Penthorum chinense Pursh-Astragalus membranaceus-Raw Rheum palmatum compound (SHY) in alleviating alcohol-induced liver injury (ALI). Methods: Network pharmacology was employed to identify active components and key targets of SHY against ALI. Protein-protein interaction (PPI) networks were constructed, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to elucidate biological processes and signaling pathways. ALI was induced in wild-type (WT) mice using the Lieber-DeCarli ethanol liquid diet (5% ethanol, vol/vol). Serum alanine aminotransferase (ALT) and interleukin-11 (IL-11) levels, along with hepatic IL-11 and phosphorylated STAT3 (p-STAT3) protein expression, were measured to validate SHY’s hepatoprotective effects. Results: Fifteen bioactive components and 219 potential targets of SHY were identified from the TCMSP database, with 106 overlapping targets linked to ALI. PPI network analysis revealed TNF, IL6, and AKT1 as core targets. GO/KEGG enrichment highlighted SHY’s regulation of oxidative stress and inflammation via IL-17 and TNF signaling pathways. Animal experiments demonstrated that SHY significantly reduced serum ALT and IL-11 levels (P < 0.01) and suppressed hepatic p-STAT3 phosphorylation. Western blot confirmed a 1.8-fold downregulation of IL-11 expression in SHY-treated mice. Conclusion: This study is the first to demonstrate that SHY ameliorates ALI by modulating the IL-11R/JAK/STAT axis, providing molecular evidence for the “treating liver diseases through intestinal regulation” theory.
基金Supported by National Natural Science Foundation of China,No. 30370438
文摘AIM： To explore the effect of recombinant human interleukin-11 （rhIL-11） on the expressions of interleukin-11 receptor α-chain （IL-11Rα） and an additional signal transducer glycoprotein 130 （gp130） in intestinal epithelium cell line-6 （IEC-6） after neutron irradiation. METHODS： Cultured IEC-6 cells were exposed to 4.0Gy neutron and treated with 100 ng/mL rhIL-11 12 h prior to or immediately after irradiation. The apoptosis and necrosis rates and expressions of IL-11Rα and gp130 were observed by flow cytometry, immunohistochemistry, Western blot and image analysis. RESULTS： The apoptosis rate of IEC-6 cells was increased by irradiation at 6 h （P 〈 0.01）, IL-11 stimulation resulted in a decreased apoptosis rate in irradiated IEC-6 cells （P 〈 0.05）. In normal control IEC-6 cells, intense immunoreactivity of IL-11Rα was located within the cell membrane and cytoplasm. The level of IL-11Rα expression significantly decreased at 6 h after irradiation （P 〈 0.01） and restored at 24 h after irradiation. In IEC-6 cells treated with both radiation and rhIL-11, the level of IL- 11Rα expression was higher than that of irradiated cells （P 〈 0.05）. When it came to gp130 protein, it was located in the cytoplasm of IEC-6 cells. After irradiation, we found a progressive decrease in the expression of gp130 protein （P 〈 0.05） in 48 hours post-radiation, while in rhIL-11-stimulated cells, it came back to normal level at 24 h after irradiation and decreased at 48 h, but was still higher than that of only irradiated cells （P 〈 0.05）. CONCLUSION： rhIL-11 can protect IEC-6 cells from neutron irradiation. The protective effect of rhIL-11 might be connected with its ability to up-regulate the expressions of specific ligand-binding subunit IL-11Rα and signal-transducing subunit gp130.
