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基于转录组测序探讨黄精芡实汤治疗糖尿病前期小鼠的作用机制

[Mechanism of Huangjing Qianshi Decoction in treatment of prediabetic mice based on transcriptome sequencing].

作者信息

Cai Jia-Luo, Zhu Yi-Lin, Li Xiao-Ping, Xia Xin-Hua, Deng Gui-Ming, Tong Qiao-Zhen, Yi Gang-Qiang, Cheng Bo

机构信息

the First Hospital Affiliated to Hunan University of Chinese Medicine Changsha 410000, China Hunan University of Chinese Medicine Changsha 410208, China.

Hunan University of Chinese Medicine Changsha 410208, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2023 Feb;48(4):1032-1042. doi: 10.19540/j.cnki.cjcmm.20220915.402.

Abstract

Based on transcriptome sequencing technology, the mouse model of prediabetes treated with Huangjing Qianshi Decoction was sequenced to explore the possible mechanism of treating prediabetes. First of all, transcriptome sequencing was performed on the normal BKS-DB mouse group, the prediabetic model group, and the Huangjing Qianshi Decoction treatment group(treatment group) to obtain differentially expressed genes in the skeletal muscle samples of mice. The serum biochemical indexes were detected in each group to screen out the core genes of Huangjing Qianshi Decoction in prediabetes. Gene Ontology(GO) database and Kyoto Encyclopedia of Genes and Genomes(KEGG) database were used to conduct signaling pathway enrichment analysis of differentially expressed genes, and real-time quantitative polymerase chain reaction(RT-qPCR) was used to verify them. The results showed that the levels of fasting blood glucose(FBG), fasting insulin(FINS), insulin resistance index(HOMA-IR), total cholesterol(TC), triglycerides(TG), and low-density lipoprotein cholesterol(LDL-C) in the mouse model were significantly decreased after treatment with Huangjing Qianshi Decoction. In the results of differential gene screening, there were 1 666 differentially expressed genes in the model group as compared with the normal group, and there were 971 differentially expressed genes in the treatment group as compared with the model group. Among them, interleukin-6(IL-6) and NR3C2 genes, which were closely related to the regulation of insulin resis-tance function, were significantly up-regulated between the model group and the normal group, and vascular endothelial growth factor A(VEGFA) genes were significantly down-regulated between the model group and the normal group. However, the expression results of IL-6, NR3C2, and VEGFA genes were adverse between the treatment group and the model group. GO functional enrichment analysis found that the biological process annotation mainly focused on cell synthesis, cycle, and metabolism; cell component annotation mainly focused on organelles and internal components; and molecular function annotation mainly focused on binding molecular functions. KEGG pathway enrichment analysis found that it involved the protein tyrosine kinase 6(PTK6) pathway, CD28-dependent phosphoinositide 3-kinase/protein kinase B(PI3K/AKT) pathway, p53 pathway, etc. Therefore, Huangjing Qianshi Decoction can improve the state of prediabetes, and the mechanism may be related to cell cycle and apoptosis, PI3K/AKT pathway, p53 pathway, and other biological pathways regulated by IL-6, NR3C2, and VEGFA.

摘要

基于转录组测序技术,对给予黄精芡实汤治疗的糖尿病前期小鼠模型进行测序,以探究治疗糖尿病前期的可能机制。首先,对正常BKS-DB小鼠组、糖尿病前期模型组和黄精芡实汤治疗组(治疗组)进行转录组测序,以获得小鼠骨骼肌样本中的差异表达基因。检测每组的血清生化指标,筛选出黄精芡实汤治疗糖尿病前期的核心基因。利用基因本体论(GO)数据库和京都基因与基因组百科全书(KEGG)数据库对差异表达基因进行信号通路富集分析,并采用实时定量聚合酶链反应(RT-qPCR)进行验证。结果显示,给予黄精芡实汤治疗后,小鼠模型的空腹血糖(FBG)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)、总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白胆固醇(LDL-C)水平均显著降低。在差异基因筛选结果中,模型组与正常组相比有1666个差异表达基因,治疗组与模型组相比有971个差异表达基因。其中,与胰岛素抵抗功能调节密切相关的白细胞介素-6(IL-6)和NR3C2基因在模型组与正常组之间显著上调,血管内皮生长因子A(VEGFA)基因在模型组与正常组之间显著下调。然而,治疗组与模型组之间IL-6、NR3C2和VEGFA基因的表达结果相反。GO功能富集分析发现,生物学过程注释主要集中在细胞合成、周期和代谢;细胞成分注释主要集中在细胞器和内部成分;分子功能注释主要集中在结合分子功能。KEGG通路富集分析发现,其涉及蛋白酪氨酸激酶6(PTK6)通路、CD28依赖性磷酸肌醇3激酶/蛋白激酶B(PI3K/AKT)通路、p53通路等。因此,黄精芡实汤可改善糖尿病前期状态,其机制可能与IL-6、NR3C2和VEGFA调控的细胞周期和凋亡、PI3K/AKT通路、p53通路等生物学途径有关。

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