基于高脂血症斑马鱼模型和网络药理学探讨黄鳝肽抗高脂血症的作用机制

doi:10.3976/j.issn.1002-4026.20230120

摘要/Abstract

摘要：

采用高血脂症斑马鱼模型和网络药理学的方法探讨黄鳝肽抗高脂血症的物质基础及作用机制。构建高血脂斑马鱼模型及测试甘油三酯染色信号强度、胆固醇荧光强度指标,分析黄鳝肽对高血脂症的改善作用;利用BIOPEP-UWM、SwissTargetPrediction、GeneCards、STRING等数据库筛选黄鳝肽及疾病靶点,使用DAVID软件进行靶点GO功能富集分析和 KEGG 通路富集分析,并构建黄鳝肽-潜在靶点-信号通路网络和蛋白互作(PPI)网络。结果显示,质量浓度为62.5、125.0 μg/mL的黄鳝肽可显著降低高脂血症斑马鱼血管中的甘油三酯染色信号强度(p<0.01)、胆固醇荧光强度(p<0.001)。网络药理学结果显示黄鳝肽中有35个潜在活性肽段序列,通过蛋白互作分析得到21个核心靶点,GO和KEGG富集分析表明黄鳝肽主要涉及脂质代谢过程的调节等生物过程,通过调节炎症、胰岛素抵抗及脂质和动脉粥样硬化等通路发挥抗高脂血症的作用。研究结果初步证实了黄鳝肽具有抗高血脂症功效,揭示了黄鳝肽降血脂生物过程中多活性肽段、多靶点、多通路的作用特点,为进一步深入研究黄鳝肽物质基础及抗高脂血症作用与应用提供理论依据及参考。

关键词: 黄鳝肽, 高脂血症, 斑马鱼, 网络药理学, 作用机制

Abstract:

This study aimed to explore the material basis and mechanism of action of Monopterus albus peptides against hyperlipidemia using hyperlipidemic zebrafish model and network pharmacology. Analysis of the ameliorating effects of Monopterus albus peptides on hyperlipidemia was conducted by constructing a hyperlipidemic zebrafish model and measuring the dye staining signal intensity of triglyceride and cholesterol fluorescence intensity changes. Monopterus albus peptides and disease targets were filtered using BIOPEP-UWM, SwissTargetPrediction, GeneCards, STRING, and other databases. Target GO functional enrichment analysis and KEGG pathway enrichment analysis were performed using the DAVID bioinformatics software, and the Monopterus albus peptides-potential target-signaling pathway network and theprotein-protein interaction (PPI) network were constructed. The results showed that Monopterus albus peptides at mass concentrations of 62.5 μg/mL and 125.0 μg/mL significantly reduced the intensity of the triglyceride staining signal (p<0.01) and cholesterol fluorescence intensity (p<0.001) in the vasculature of hyperlipidemic zebrafish. The results of network pharmacology showed that there were 35 potentially active peptide sequences in Monopterus albus peptides, and 21 core targets were obtained by protein interaction analysis. GO and KEGG enrichment analyses indicated that Monopterus albus peptides were mainly involved in biological processes such asregulation of lipid metabolic processes, and exertan antihyperlipidemia effect viainflammation regulation, insulin resistance, and lipid and atherosclerosis channels. Initially, this study confirmed that Monopterus albus peptides have an antihyperlipidemia effect, and revealed that Monopterus albus peptides have multiple active peptide fragments, multiple targets, and multiple channels in the biological process of reducing blood lipid levels, which will provide a theoretical basis and reference for further in-depth research on the material basis of Monopterus albus peptides and their role and application in countering hyperlipidemia.

Key words: Monopterus albus peptides, hyperlipidemia, zebrafish, network pharmacology, mechanism of action

中图分类号:

R285

马诗经, 何春艳, 关天竹, 姚雪霜, 张俊鹏. 基于高脂血症斑马鱼模型和网络药理学探讨黄鳝肽抗高脂血症的作用机制[J]. 山东科学, 2024, 37(3): 27-38.

MA Shijing, HE Chunyan, GUAN Tianzhu, YAO Xueshuang, ZHANG Junpeng. Exploration of antihyperlipidemia mechanism of Monopterus albus peptides based on hyperlipidemic zebrafish model and network pharmacology[J]. Shandong Science, 2024, 37(3): 27-38.

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参考文献 21

[1]	CHEN H, MIAO H, FENG Y L, et al. Metabolomics in dyslipidemia[J]. Advances in Clinical Chemistry, 2014, (66): 101-119.
[2]	诸骏仁, 高润霖, 赵水平, 等. 中国成人血脂异常防治指南(2016年修订版)[J]. 中国循环杂志, 2016, 31(10): 937-953.
[3]	吴敏, 冯玲, 沈冬. 高脂血症药物治疗研究进展[J]. 中国临床医生, 2012, 40(3): 35-37.
[4]	韩林, 丁博, 王兆丹, 等. 黄鳝肉双酶水解产物的功能特性及其抗氧化活性[J]. 食品与发酵工业, 2014, 40(10): 151-155. DOI: 10.13995/j.cnki.11-1802/ts.2014.10.065.
[5]	姚晓燕. 黄鳝骨硫酸软骨素多糖降血脂功能研究[J]. 中国医药导报, 2011, 8(30): 31-33. DOI: 10.3969/j.issn.1673-7210.2011.30.014.
[6]	关天竹, 何春艳, 王涛, 等. 基于斑马鱼模型的黄鳝肽减肥作用研究[J]. 食品安全导刊, 2022, (33): 95-98. DOI: 10.16043/j.cnki.cfs.2022.33.020.
[7]	李联泰, 安贤惠, 胡江, 等. 黄鳝皮肤黏液抗菌肽的分离纯化及其部分特性研究[J]. 渔业科学进展, 2011, 32(2): 27-33. DOI: 10.3969/j.issn.1000-7075.2011.02.005.
[8]	李心怡, 陈荷清, 夏欢, 等. 蜂胶降脂方对高脂血症模型斑马鱼的降血脂作用研究[J]. 世界科学技术—中医药现代化, 2020, 22(5): 1629-1635.
[9]	黄帅, 成鹏, 杨宇, 等. 基于网络药理学探究蒲参胶囊治疗高脂血症的作用机制[J]. 南京中医药大学学报, 2019, 35(3): 290-296. DOI: 10.14148/j.issn.1672-0482.2019.0290.
[10]	CHOUNG S, KIM J M, JOUNG K H, et al. Epidermal growth factor receptor inhibition attenuates non-alcoholic fatty liver disease in diet-induced obese mice[J]. PLoS One. 2019, 14(2): e0210828. DOI: 10.1371/journal.pone.0210828.
[11]	钟华, 仇静文, 吴鸿飞, 等. 基于网络药理学研究瓜蒌—薤白药对抗高脂血症作用机制[J]. 中国实验方剂学杂志, 2020, 26(18): 154-165. DOI: 10.13422/j.cnki.syfjx.20201603.
[12]	OKWAN-DUODU D, WEISS D, PENG Z, et al. Overexpression of myeloid angiotensin-converting enzyme (ACE) reduces atherosclerosis[J]. Biochemical and Biophysical Research Communications, 2019, 520(3): 573-579. DOI: 10.1016/j.bbrc.2019.10.078.
[13]	邹振武, 李德忠, 彭绪东, 等. 单味中药鬼针草颗粒治疗高脂血症的疗效及其对血清MMP-9、TIMP-1水平的影响[J]. 心血管康复医学杂志, 2019, 28(5): 661-665. DOI: 10.3969/j.issn.1008-0074.2019.05.25.
[14]	YAN H, WANG J, FU H, et al. Discussion on the potential target and mechanism of Dachaihu Decoction in treating hyperlipidemia based on network pharmacology[J]. Journal of Chinese Pharmaceutical Sciences, 2023, 32(6): 446-459. doi: 10.5246/jcps.2023.06.038
[15]	KIMY U, KEE P, DANILA D, et al. A critical role of PCSK9 in mediating IL-17-producing T cell responses in hyperlipidemia[J]. Immune Network, 2019, 19(6): e41. DOI: 10.4110/in.2019.19.e41.
[16]	HE Y H, YANG G D, SUN L J, et al. SIRT6 inhibits inflammatory response through regulation of NRF₂ in vascular endothelial cells[J]. International Immunopharmacology, 2021, 99: 107926. DOI: 10.1016/j.intimp.2021.107926.
[17]	UEKITA H, ISHIBASHI T, SHIOMI M, et al. Integral role of receptor for advanced glycation end products (RAGE) in nondiabetic atherosclerosis[J]. Fukushima Journal of Medical Science, 2019, 65(3): 109-121. DOI: 10.5387/fms.2019-12. pmid: 31915324
[18]	YE J H, LI L, HU Z X. Exploring the molecular mechanism of action of Yinchen Wuling Powder for the treatment of hyperlipidemia, using network pharmacology, molecular docking, and molecular dynamics simulation[J]. BioMed Research International, 2021, 2021: 1-14. DOI: 10.1155/2021/9965906.
[19]	柴露露. 银杏叶制剂治疗高脂血症的疗效及作用机制研究[D]. 北京: 中国中医科学院, 2019.
[20]	李文毅, 周春阳. 高脂血症与动脉粥样硬化和脂代谢研究进展[J]. 中国药理学与毒理学杂志, 2019, 33(10): 811.
[21]	WU X Z, PAN J X, YU J, et al. DiDang decoction improves mitochondrial function and lipid metabolism via the HIF-1 signaling pathway to treat atherosclerosis and hyperlipidemia[J]. Journal of Ethnopharmacology, 2023, 308: 116289. DOI: 10.1016/j.jep.2023.116289.

实验组	质量浓度/(μg·mL^-1)	死亡数/尾	死亡率/%	表型
正常对照组	未添加	0	0	未见明显异常
	62.5	0	0	与正常对照组状态相似
	125.0	0	0	与正常对照组状态相似
黄鳝肽	250.0	30	100	—
	500.0	30	100	—
	1 000.0	30	100	—

实验组	质量浓度/(μg·mL^-1)	尾部血管染色强度/像素	甘油三酯抑制率/%
正常对照组	未添加	459±39^***
模型对照组	未添加	13 464±814
阿伐他汀钙组	11.6	4 050±294^***	69.9
黄鳝肽低剂量组	31.2	11 732±399	12.9
黄鳝肽中剂量组	62.5	6 915±234^***	48.6
黄鳝肽高剂量组	125.0	7 351±374^***	45.4

实验组	质量浓度/(μg·mL^-1)	尾部血管胆固醇荧光强度/像素	胆固醇抑制率/%
正常对照组	未添加	15 640±946^***
模型对照组	未添加	47 552±3 555
阿伐他汀钙组	11.6	33 087±2 906^**	30.4
黄鳝肽低剂量组	31.2	35 188±2 935	26.0
黄鳝肽中剂量组	62.5	29 848±2 422^**	37.2
黄鳝肽高剂量组	125.0	28 914±2 370^**	39.2

肽段序列	肽活性评分	血脑屏障通透性	分子疏水常数	生物利用度评分
PGPLGPYFL	0.96	NO	-1.39	0.17
PGPM	0.95	NO	-0.77	0.55
FSAPGGGF	0.95	—	-2.91	0.17
PGPGMP	0.94	—	-1.60	0.17
PGPLGPM	0.93	—	-1.55	0.17
PGPGPM	0.93	—	-1.60	0.17
GPPVPGPLGP	0.93	—	-2.75	0.17
PDPGPGPM	0.93	—	-2.83	0.11
LPGPGACGKF	0.92	—	-3.51	0.17
PGPLGPL	0.92	—	-1.36	0.17
PDPGPGMP	0.92	—	-2.83	0.11
PGLPGPM	0.91	—	-1.55	0.17
PGPLGMP	0.91	—	-1.55	0.17
PGLPGACGKF	0.91	—	-3.51	0.17
GPPSGGFK	0.90	—	-3.60	0.17
PVPGLPGPM	0.90	—	-2.00	0.17
SSGPPVPGPL	0.88	—	-4.34	0.17
PGPLGPMT	0.88	—	-2.66	0.17
PGLPCGAGKF	0.87	—	-3.51	0.17
AGGPLGPR	0.86	—	-3.51	0.17
PVPGPM	0.85	—	-0.98	0.17
VPGLPGPM	0.85	—	-1.76	0.17
VPPGPM	0.85	—	-0.98	0.17
GPAPP	0.85	—	-1.29	0.55
GPGR	0.84	—	-2.42	0.17
WDPGPQ	0.84	—	-2.72	0.11
GPAGSFGPVGK	0.84	—	-5.12	0.17
PGGGFDL	0.84	—	-2.50	0.11
PGLPGP	0.84	—	-1.39	0.17
GPAGPHGPVG	0.83	—	-4.80	0.17
PAGPL	0.82	—	-1.06	0.55
SGPPAP	0.82	—	-2.55	0.17
GPAGGPAGSF	0.81	—	-4.71	0.17
GPVGPM	0.81	—	-1.60	0.17
GPAGPR	0.80	—	-2.88	0.17

核心靶点名称	中间度值	接近度值	节点度值	核心靶点名称	中间度值	接近度值	节点度值
TNF	1 094.86	0.009 3	54	DPP4	220.39	0.006 6	16
EGFR	444.02	0.008 1	40	CASP3	212.92	0.008 1	38
PPARA	385.85	0.007 2	25	PLG	196.49	0.007 2	27
ACE	336.95	0.007 7	32	FOS	176.27	0.007 7	33
REN	284.97	0.007 3	29	MMP2	139.78	0.007 5	32
F2	278.27	0.006 9	22	SERPINE1	139.33	0.007 4	28
APP	263.44	0.007 3	27	PIK3CA	120.40	0.006 4	23
CCND1	254.93	0.007 6	33	SIRT1	108.19	0.007 4	28
HSP90AA1	249.20	0.007 2	29	MAPK8	102.92	0.007 4	30
MMP9	232.55	0.008 3	41	PTGS2	93.59	0.007 7	34
PPARG	227.57	0.007 6	32