基于转录组学技术研究泽泻汤对高脂血症大鼠的降脂作用机制

doi:10.5246/jcps.2021.05.032

中国药学(英文版) ›› 2021, Vol. 30 ›› Issue (5): 409-420.DOI: 10.5246/jcps.2021.05.032

基于转录组学技术研究泽泻汤对高脂血症大鼠的降脂作用机制

徐小妹¹, 李丽莎¹, 张亚敏¹, 卢雪花¹, 徐榕青¹, 吴双双¹, 林纬², 林文津¹^,^*()

1. 福建省医学科学研究院福建省医学测试重点实验室, 福建福州 350001
2. 福建省立医院, 福建福州 350001

收稿日期:2020-09-26 修回日期:2020-10-26 接受日期:2020-11-19 出版日期:2021-05-31 发布日期:2021-05-29
通讯作者: 林文津
作者简介:
+ Tel.: +86-591-87514999, E-mail: Lwj680@139.com
基金资助:
The Basic Research Project of Fujian Provincial Public Welfare Research Institute (Grant No. 2018R1031-3),?Medical Innovation Project of Fujian Provincial Health Commission (Grant No. 2018-CX-15), Basic Research Project of Fujian Provincial Public Welfare Research Institute, China (Grant No. 2019R1011-5) and High-Level?Hospital?grants (Grant No. 2017GL-001) from?Fujian Provincial Hospital, Fujian Province, China.

Hypolipidemic effects of Alismatis Rhizoma decoction on the lipid profile in hyperlipidemia rats by RNA-sequencing

Xiaomei Xu¹, Lisha Li¹, Yamin Zhang¹, Xuehua Lu¹, Rongqing Xu¹, Shuangshuang Wu¹, Wei Lin², Wenjin Lin¹^,^*()

1 Fujian Key Laboratory of Medical Measurement, Fujian Academy of Medical Sciences, Fuzhou 350001, China
2 Department of Endocrinology, Shengli Clinical Medical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou 350001, China

Received:2020-09-26 Revised:2020-10-26 Accepted:2020-11-19 Online:2021-05-31 Published:2021-05-29
Contact: Wenjin Lin

摘要/Abstract

摘要：

高脂血症是一种脂质代谢紊乱, 其特征是血液循环中脂质水平升高。高脂血症一直与心血管事件和代谢综合征相关疾病的发展有关。泽泻汤(ARD)是著名的中药处方, 长期以来被用于治疗眩晕症, 这是高脂血症患者的一种症状。在本研究中, 我们旨在转录水平上研究ARD对高脂血症大鼠的降脂活性和潜在分子机制。同时应用RNA测序和转录组分析, 包括差异表达基因(DEGs)分析、GO功能和KEGG通路分析。结果表明, 与CON组相比, HFD组中识别出1981个DEGs(上调1370个, 下调611个)。与HFD组相比, ARD组检测到474个DEGs(350个上调和124个下调)。此外, GO分析表明DEGs主要参与以下功能: 发育过程、对外部刺激的反应、离子迁移、酒精结合和质膜部分。路径分析表明, 这些DEGs显著存在于在胆汁分泌、疟疾、细胞粘附分子、视黄醇代谢、鞘脂信号传导途径、化学致癌作用和T细胞受体信号传导途径。研究表明ARD通过多种机制减轻了高脂血症引起的脂质代谢紊乱, 这为ARD的进一步药理研究提供了重要的科学依据。

关键词: 泽泻汤, 高脂血症, RNA测序, 脂质

Abstract:

As a disorder of lipid metabolism, hyperlipidemia (HLP) is characterized by elevated levels of lipids in the blood circulation. It is consistently related to the development of cardiovascular events and diseases associated with metabolic syndrome. Alismatis Rhizoma decoction (ARD), a well-known traditional Chinese medicine prescription, has long been used for treating vertigo, which is a symptom experienced by HLP patients. In this study, we aimed to investigate the hyperlipidemic activity and the potential molecular mechanisms of ARD in HLP rats at the transcriptional level. RNA sequencing and transcriptome analysis were performed collaboratively, including analysis of differentially expressed genes (DEGs), GO functions, and KEGG pathway analysis. The results showed that 1981 DEGs (1370 upregulated and 611 downregulated) were identified in the HFD group compared with the CON group. Moreover, 474 DEGs (350 upregulated and 124 downregulated) were detected in the ARD group compared with the HFD group. Furthermore, GO analysis revealed that DEGs were mainly involved in the following functions: developmental process, response to an external stimulus, ion transport, alcohol binding, and plasma membrane part. Pathway analysis suggested that these DEGs were significantly enriched in bile secretion, malaria, cell adhesion molecules, retinol metabolism, the sphingolipid signaling pathway, chemical carcinogenesis, and the T cell receptor signaling pathway. In conclusion, our study demonstrated that ARD alleviated the lipid metabolism disorder caused by HLP through multiple mechanisms, which provided vital scientific evidence for further pharmacological studies of ARD.

Key words: Alismatis Rhizoma decoction, Hyperlipidemia, RNA-sequencing, Lipid profile

Supporting:

徐小妹, 李丽莎, 张亚敏, 卢雪花, 徐榕青, 吴双双, 林纬, 林文津. 基于转录组学技术研究泽泻汤对高脂血症大鼠的降脂作用机制[J]. 中国药学(英文版), 2021, 30(5): 409-420.

Xiaomei Xu, Lisha Li, Yamin Zhang, Xuehua Lu, Rongqing Xu, Shuangshuang Wu, Wei Lin, Wenjin Lin. Hypolipidemic effects of Alismatis Rhizoma decoction on the lipid profile in hyperlipidemia rats by RNA-sequencing[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(5): 409-420.

图/表 6

Figure 1. Effects of ARD on the levels of serum biochemical parameters. (A) The serum level of TC; (B) The serum level of TG; (C) The serum level of LDL-C; (D) The serum level of HDL-C. Data represent the mean ± SEM (n = 8). *P < 0.05 vs. CON group, #P < 0.05 vs. HFD group.

Figure 2. ARD administration attenuates HFD-induced liver injury. (A) The serum levels of ALT and AST; (B) The gross appearance of rat livers; (C) The histopathological observation of rat livers. H&E stained sections were photographed at 200× magnification. Data represent the mean ± SEM (n = 8). *P < 0.05 vs. CON group, #P < 0.05 vs. HFD group.

Figure 3. Gene expression in livers of HLP rats detected by RNA-sequencing. (A) Heatmap for hierarchical cluster analysis of DEGs. Colors stand for the log intensities of gene expression levels with red indicating high expression and green indicating low expression. Genes (|log2FoldChange| > 1 and an FDR < 0.05) are analyzed in the heatmap; (B) The number of significantly upregulated and downregulated genes in CON_vs._HFD, CON_ vs._ARD, and HFD_vs._ARD; (C) Venn diagram of the counts of DEGs in two comparisons for liver tissues (CON_ vs._HFD, HFD_vs._ARD).

Figure 4. Functional enrichment analysis of co-expressed DEGs in the livers of HLP rats. (A) GO term of co-expressed DEGs in CON_vs._HFD and HFD_vs._ARD; (B) Pathway functional enrichment of co-expressed DEGs in CON_ vs._HFD and HFD_vs._ARD. (C) The FPKM values of some important genes.

Table 1. Some typical genes involved in the cellular lipid metabolic pathways, which were regulated by HFD consumption and counteracted by ARD supplementation.

Figure 5. The expression levels of the Mt1, Sat1, and Gstm3_1 were verified by real-time PCR (n = 3). *P < 0.05 vs. CON group, #P < 0.05 vs. HFD group.

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基于转录组学技术研究泽泻汤对高脂血症大鼠的降脂作用机制

Hypolipidemic effects of Alismatis Rhizoma decoction on the lipid profile in hyperlipidemia rats by RNA-sequencing

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