Metabolomic Profiling Reveals Protective Effects and Mechanisms of Sea Buckthorn Sterol against Carbon Tetrachloride-Induced Acute Liver Injury in Rats
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animals, Reagents, and Drugs
2.2. Preparations of the Acute Liver Injury Model
2.3. Pathological Examination of Liver Tissue
2.4. Measurement of Biochemical Index in Liver Tissue Homogenate
2.5. Sample Preparation
2.5.1. Extraction of Hydrophilic Compounds
2.5.2. Extraction of Hydrophobic Compounds
2.6. Analysis by UPLC
2.7. Analysis by ESI-Q TRAP-MS/MS
2.7.1. ESI-Q TRAP-MS/MS of Hydrophilic Compounds
2.7.2. ESI-Q TRAP-MS/MS of Hydrophobic Compounds
2.8. Transcriptome Analysis
2.8.1. RNA Detection
2.8.2. Library Setup
Library Quality Check
- (i)
- Qubit2.0 was used for preliminary quantification, and Agilent 2100 was used to detect the insert size of the library. The next experiment could only be carried out after the insert size met expectations;
- (ii)
- The Q-PCR method accurately quantified the effective concentration of the library (the effective concentration of the library was >2 nM). Then, the library check was successfully performed.
Sequencing on the Machine
2.9. Statistical Analysis
3. Results
3.1. Effects of SBS on SOD, MDA, and GSH-PX in Rats with CCl4-Induced Liver Injury
3.2. Effects of SBS on CAT and T-AOC in Rats with CCl4-Induced Liver Injury
3.3. Effects of SBS on γ-GT, TP, PGE2, and COX-2 in Rats with CCl4-Induced Liver Injury
3.4. Electron Microscopic Observations of Liver Tissue
3.5. Light Microscopic Observations of Liver Tissue
3.6. Identification and Analysis of Metabolites
3.6.1. Discriminant Analysis of Orthogonal Partial Least Squares Method
3.6.2. Determination of Relevant Metabolite
3.6.3. Metabolic Pathway Enrichment Analysis
3.7. Identification and Selection of Differential Genes
3.7.1. Differential Gene Volcano Map
3.7.2. Expression of Key Genes
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Group | Results |
---|---|
BG | (−) |
MG | Feathery degeneration of hepatocyte (++) Fatty degeneration of hepatocytes (++) Hepatocyte necrosis (++) Inflammatory cell infiltration (+) |
DDB | A small amount of feathery degeneration of hepatocytes (+) A small amount of fatty degeneration of hepatocytes (+) Hepatocyte necrosis (+) |
LD | Feathery degeneration of hepatocytes (++) Fatty degeneration of hepatocytes (+) Hepatocyte necrosis (+) Inflammatory cell infiltration (+) |
MD | Feathery degeneration of hepatocytes (++) Fatty degeneration of hepatocytes (+) Hepatocyte necrosis (++) Inflammatory cell infiltration (+) |
HD | A small amount of feathery degeneration of hepatocytes (+) A small amount of fatty degeneration of hepatocytes (+) Hepatocyte necrosis (+) |
NO. | Metabolites | RT | MG/BG | LD/MG | MD/MG | HD/MG | DDB/MG | Pathway |
---|---|---|---|---|---|---|---|---|
1 | L-Malic Acid | 0.88 | ↑ * | ↓/- | ↓ * | ↓ * | ↓/- | Citrate cycle (TCA cycle) |
2 | 7Z, 10Z, 13Z, 16Z, 19Z-docosapentaenoic acid | 11.3 | ↑ * | ↓ * | ↓ * | ↓ * | ↓/- | Biosynthesis of unsaturated fatty acids |
3 | creatine | 0.78 | ↑ * | ↓/- | ↓ * | ↓ * | ↓ * | Arginine-proline metabolism |
4 | n-acetyl-l-alanine | 1.3 | ↑ * | ↓/- | ↓ * | ↓ * | ↓ * | - |
5 | N-Acetylaspartate | 0.69 | ↑ * | ↓/- | ↓ * | ↓ * | ↓ * | Alanine, aspartate and glutamate metabolism |
6 | Trigonelline | 0.77 | ↑ * | ↓/- | ↓ * | ↓ * | ↓/- | Nicotinate and nicotinamide metabolism |
7 | 4-guanidinobutyric acid | 0.85 | ↑ * | ↓ * | ↓ * | ↓ * | ↓ * | Arginine-proline metabolism |
8 | N-Amidino-L-Aspartate | 0.96 | ↑ * | ↓ * | ↓ * | ↓ * | ↓ * | - |
9 | n-glycyl-l-leucine | 1.8 | ↓ * | ↑/- | ↑ * | ↑ * | ↑/- | - |
10 | FFA(6:0) | 0.73 | ↓ * | ↑ * | ↑ * | ↑ * | ↑ * | Fat digestion and absorption |
11 | CE(16:1) | 13.12 | ↑ * | ↓ * | ↓ * | ↓ * | ↓ * | - |
12 | CE(18:2) | 13.21 | ↑ * | ↓ * | ↓ * | ↓ * | ↓ * | - |
13 | PE(16:1/16:0) | 6.25 | ↑ * | ↓ * | ↓ * | ↓ * | ↓ * | Glycerophospholipid metabolism |
14 | DG(16:0/20:2/0:0) | 9.18 | ↑ * | ↓/- | ↓ * | ↓ * | ↓/- | Glycerolipid metabolism |
15 | TG(14:0/18:0/18:2) | 11.85 | ↑ * | ↓/- | ↓ * | ↓ * | ↓ * | Glycerolipid metabolism |
16 | TG(14:0/18:0/20:4) | 11.81 | ↑ * | ↓/- | ↓ * | ↓ * | ↓ * | Glycerolipid metabolism |
17 | TG(16:0/16:1/22:5) | 11.41 | ↑ * | ↓/- | ↓ * | ↓ * | ↓ * | Glycerolipid metabolism |
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Sheng, C.; Guo, Y.; Ma, J.; Hong, E.-K.; Zhang, B.; Yang, Y.; Zhang, X.; Zhang, D. Metabolomic Profiling Reveals Protective Effects and Mechanisms of Sea Buckthorn Sterol against Carbon Tetrachloride-Induced Acute Liver Injury in Rats. Molecules 2022, 27, 2224. https://doi.org/10.3390/molecules27072224
Sheng C, Guo Y, Ma J, Hong E-K, Zhang B, Yang Y, Zhang X, Zhang D. Metabolomic Profiling Reveals Protective Effects and Mechanisms of Sea Buckthorn Sterol against Carbon Tetrachloride-Induced Acute Liver Injury in Rats. Molecules. 2022; 27(7):2224. https://doi.org/10.3390/molecules27072224
Chicago/Turabian StyleSheng, Changting, Yang Guo, Jing Ma, Eun-Kyung Hong, Benyin Zhang, Yongjing Yang, Xiaofeng Zhang, and Dejun Zhang. 2022. "Metabolomic Profiling Reveals Protective Effects and Mechanisms of Sea Buckthorn Sterol against Carbon Tetrachloride-Induced Acute Liver Injury in Rats" Molecules 27, no. 7: 2224. https://doi.org/10.3390/molecules27072224