Molecular mechanism of high-altitude hypoxia-induced lipid metabolism disorder in mouse spleen tissue

doi:10.12122/j.issn.1673-4254.2024.10.21

Abstract

Abstract:

Objective To investigate the molecular mechanism of lipid metabolism disorder in mouse spleen tissues due to high-altitude hypoxia. Methods Ten C57BL/6 male mice were randomly divided into normoxia group (maintained at an altitude of 400 m) and high-altitude hypoxia group (maintained at 4200 m) for 30 days (n=5). Lipidomics and metabolomics analyses of the spleen tissue of the mice were conducted using liquid chromatography-mass spectrometry (LC-MS) to identify the differential metabolites, which were further analyzed by KEGG enrichment and pathway analyses, and the differential genes were screened through transcriptome sequencing. Bioinformatics analysis was conducted to identify the upstream target genes of the differential metabolites in specific metabolic pathways. RT-qPCR and Western blotting were used to detect mRNA expressions of 11β‑hydroxysteroid dehydrogenase 1 (HSD11B1), steroid 5α reductase 1 (SRD5A1), prostaglandin-endoperoxide synthase 1 (PTGS1), hematopoietic prostaglandin D synthetase (HPGDS), xanthine dehydrogenase (XDH), purine nucleoside phosphorylase (PNP), hypoxanthine guanine-phosphoribosyltransferase (HPRT) and extracellular 5'-nucleotidase (NT5E) and protein expressions of HSD11B1, SRD5A1, XDH, PNP and HPRT in the mouse spleens. Results We identified a total of 41 differential lipid metabolites in the mouse spleens, and these metabolites and the differential genes were enriched in steroid hormone biosynthesis, arachidonic acid metabolism, and purine metabolism pathways. Compared to the mice kept in normoxic conditions, the mice exposed to high-altitude hypoxia showed significantly upregulated expressions of adrenosterone, androsterone, prostaglandin D2, prostaglandin J2, xanthine, xanthosine, and uric acid in the spleen with also changes in the expression levels of HSD11B1, SRD5A1, PTGS1, HPGDS, XDH, PNP, HPRT, and NT5E. Conclusion High-altitude hypoxia can result in lipid metabolism disorder in mouse spleen tissue by affecting steroid hormone biosynthesis, arachidonic acid metabolism, and purine metabolism pathways.

Key words: lipid metabolism, high-altitude hypoxia, metabolomics, transcriptomics, spleen, lipidomics

Chengling CUI, Yuzhen XU, Chaoqun TANG, Jiaying JIANG, Ying HU, Jie SHUANG. Molecular mechanism of high-altitude hypoxia-induced lipid metabolism disorder in mouse spleen tissue[J]. Journal of Southern Medical University, 2024, 44(10): 2024-2032.

Figures/Tables 12

Tab.1 RT-qPCR primer sequences

Gene	Primer sequence (5′→3′)	bp
β-Actin	F: CATCCGTAAAGACCTCTATGCCAAC	25
	R: ATGGAGCCACCGATCCACA	19
HSD11B1	F: GCCAGGTCGGAGGAAGGTC	19
	R:GCCAGCAATGTAGTGAGCAGAG	22
SRD5A1	F: TCGTGGAGTGGTGTGGCTTTG	21
	R: TGATGATGCTGCCTCGCTCTG	21
PTGS1	F: TGTTGTTACTATCCGTGCCAGAAC	24
	R: TTCCGAAGCCAGGTCCAGATC	21
HPGDS	F: TGGTGGACACGCTGGATGAC	20
	R: CAGAAGGCGAGGTGCTTGATG	21
XDH	F: TTGCGAAGGATGAGGTTACTTGTG	24
	R: CTGGATTGTGATAATGGCTGGAAGG	25
NT5E	F: GACCAGTACCAGGGCACCATC	21
	R: CAATCAGTCCTTCCACACCGTTATC	25
HPRT	F: AGTCCCAGCGTCGTGATTAGC	21
	R: CGAGCAAGTCTTTCAGTCCTGTC	23
PNP	F: CAAAGGTGACATTCCCAGTGAGAG	24
	R: GTTGAGTCCTCCAGCAGCATTG	22

Tab.2 Detection of spleen index of the mice in the two groups (Mean±SD)

Group	Body weight (g)	Spleen quality (g)	Spleen index (%)
PSC	22.890±0.265	0.0632±0.002	0.276±0.009
HST	20.280±0.347	0.0491±0.004	0.240±0.023
t	11.97****	6.722***	2.879*

Fig.1 Analysis of the lipidomics data of the mice. A: PCA analysis. B: Volcano map. C: Type and number of the enriched lipid metabolites. D: Correlation analysis of the differential lipids.

Tab.3 Number of the enriched differential metabolites

Compared samples

Total metabolite identification results

Total number of metabolites

with significant differences

Significantly

upregulated metabolites

Significantly downregulated metabolites

Fig.2 Z-score plot.

Fig.3 KEGG pathway diagram. A: KEGG annotation analysis plot. B: KEGG enrichment bubble diagram.

Fig.4 LIPID MAPS classification annotation.

Fig.5 Transcriptomics data analysis. A: Volcano map. B: KEGG pathway diagram.

Fig.6 Transcriptional metabolism combined with KEGG pathway analysis.

Fig.7 Relative abundance of the differential metabolites in lipid metabolism-related pathways. *P<0.05, **P<0.01, ***P<0.001.

Fig.8 Expression of the key genes in lipid metabolism-related pathways. *P<0.05, **P<0.01,***P<0.001.

Fig.9 Protein expression of the genes in lipid metabolism-related pathways in the mouse spleens detected by Western blotting. A: Protein bands. B: Relative protein expression levels. *P<0.05, **P<0.01, ***P<0.001.

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