返回 
加入收藏
基于转录组学和蛋白质组学研究银杏黄酮苷元减轻多柔比星心脏毒性的作用机制
x
请在关注微信后，向客服人员索取文件
| 篇名： | 基于转录组学和蛋白质组学研究银杏黄酮苷元减轻多柔比星心脏毒性的作用机制 |
| TITLE： | Mechanism of Ginkgo flavone aglycone in alleviating doxorubicin-induced cardiotoxicity based on transcriptomics and proteomics |
| 摘要： | 目的 基于转录组学和蛋白质组学研究银杏黄酮苷元（GA）降低多柔比星（DOX）心脏毒性的作用机制。方法将36只小鼠随机分为对照组（CON组，隔天尾静脉注射等体积生理盐水+每天灌胃等体积生理盐水）、DOX组（隔天尾静脉注射3mg/kgDOX）和GDOX组（每天灌胃100mg/kgGA+隔天尾静脉注射3mg/kgDOX），每组12只，连续给药/生理盐水15d。收集小鼠心脏组织进行RNA-Seq转录组学测序和4D-Label-free定量蛋白质组学分析，筛选差异表达基因和蛋白并对其进行京都基因与基因组百科全书（KEGG）富集分析。测定小鼠心脏组织中爱帕琳肽（Apelin）、磷脂酰肌醇3-激酶（PI3K）和蛋白激酶B（Akt）的mRNA和蛋白表达水平及PI3K、Akt蛋白的磷酸化水平进行验证。将H9c2心肌细胞分为对照组（CON组）、DOX组（2μmol/L）和GDOX组（2μg/mLGA+2μmol/LDOX），测定细胞存活率及细胞中糖酵解关键物质水平。结果筛选得到6条转录组学与蛋白质组学的共同通路，包括Apelin信号通路、PI3K-Akt信号通路、胰岛素抵抗等；其中以Apelin、PI3K-Akt信号通路中富集到的基因数目最多。靶点验证实验结果显示，与CON组比较，DOX组小鼠心脏组织中Apelin、PI3K和Akt的mRNA相对表达量和蛋白表达水平，以及PI3K、Akt蛋白的磷酸化水平均显著降低（P＜0.05或P＜0.01）；与DOX组比较，GDOX组小鼠心脏组织中Apelin、PI3K和Akt的mRNA相对表达量及PI3K、Akt蛋白的磷酸化水平均显著升高（P＜0.05或P＜0.01）。细胞实验表明，与CON组比较，DOX组细胞存活率显著降低（P＜0.05），细胞中葡萄糖相对摄取量以及丙酮酸、乳酸的相对生成量均显著增加（P＜0.05），ATP相对生成量显著减少（P＜0.05）；与DOX组比较，GDOX组细胞存活率显著升高（P＜0.05），细胞中丙酮酸、乳酸的相对生成量均显著减少（P＜0.05）。结论GA可能是通过上调心脏组织中Apelin、PI3K和Akt的mRNA及其蛋白表达，调控糖酵解过程，进而改善DOX引起的心脏毒性作用。 |
| ABSTRACT： | OBJECTIVE To investigate the mechanism by which Ginkgo flavone aglycone （GA） reduces the cardiotoxicity of doxorubicin （DOX） based on transcriptomics and proteomics. METHODS Thirty-six mice were randomly assigned to control group （CON group， tail vein injection of equal volume of physiological saline every other day+daily intragastric administration of an equal volume of physiological saline）， DOX group （tail vein injection of 3 mg/kg DOX every other day）， and GDOX group （daily intragastric administration of 100 mg/kg GA+tail vein injection of 3 mg/kg DOX every other day）， with 12 mice in each group. The administration of drugs/physiological saline was continued for 15 days. Mouse heart tissues were collected for RNA-Seq transcriptomic sequencing and 4D-Label-free quantitative proteomic analysis to screen differentially expressed genes and proteins， which were then subjected to Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis. The expression levels of Apelin peptide （Apelin）， phosphatidylinositol 3-kinase （PI3K）， and protein kinase B （Akt） mRNA and protein in mouse heart tissues， as well as the phosphorylation levels of PI3K and Akt proteins， were verified. H9c2 cardiomyocytes were divided into control group （CON group）， DOX group （2 μmol/L）， and GDOX group （2 μg/mL GA+2 μmol/L DOX） to determine cell viability and the levels of key glycolytic substances in the cells. RESULTS Six common pathways were identified from transcriptomics and proteomics， including the Apelin signaling pathway， the PI3K-Akt signaling pathway， and insulin resistance. Among them， the Apelin and PI3K-Akt signaling pathways were the most enriched in terms of gene numbers. Target validation experiments showed that compared to the CON group， the relative expression of Apelin， PI3K and Akt mRNA and protein levels， as well as the phosphorylation levels of PI3K and Akt proteins， were significantly decreased in the DOX group （P＜0.05 or P＜0.01）. The relative expression of Apelin， PI3K and Akt mRNA and the phosphorylation levels of PI3K and Akt proteins were significantly increased in the GDOX group as compared with the DOX group （P＜0.05 or P＜0.01）. Cellular experiments indicated that compared to the CON group， cell viability in the DOX group was significantly decreased （P＜0.05）， the relative uptake of glucose and the relative production of pyruvate and lactate were significantly increased （P＜0.05）， and the relative production of ATP was significantly reduced （P＜0.05）. Compared to the DOX group， cell viability in the GDOX group was significantly increased （P＜ 0.05）， and the relative production of pyruvate and lactate was significantly reduced （P＜0.05）. CONCLUSIONS GA may alleviate DOX-induced cardiotoxicity by upregulating the mRNA and protein expression of Apelin， PI3K， and Akt in heart tissues， and regulating glycolytic processes. |
| 期刊： | 2024年第35卷第21期 |
| 作者： | 涂玉洁；蔡英；程雪怡；孙佳；潘洁；刘春花；李勇军；黄勇；郑林；陆苑 |
| AUTHORS： | TU Yujie， CAI Ying，CHENG Xueyi，SUN Jia，PAN Jie，LIU Chunhua，LI Yongjun，HUANG Yong，ZHENG Lin，LU Yuan |
| 关键字： | 银杏黄酮苷元；多柔比星；心脏毒性；转录组学；蛋白质组学；减毒机制 |
| KEYWORDS： | Ginkgo flavone aglycone； doxorubicin； cardiotoxicity； transcriptomics； proteomics； toxicity reduction mechanism |
| 阅读数： | 5 次 |
| 本月下载数： | 0 次 |
* 注：未经本站明确许可，任何网站不得非法盗链资源下载连接及抄袭本站原创内容资源!在此感谢您的支持与合作！
