Journal of Traditional Chinese Medicine ›› 2023, Vol. 43 ›› Issue (2): 274-285.DOI: 10.19852/j.cnki.jtcm.20230201.001
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Liver metabolomic characteristics in three different rat models of Yin deficiency based on ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry
HU Xingyao, LIU Hongning, YAN Xiaojun, CHEN Zhong, FU Liu, LIU Ge, CHEN Xuan, SHANG Guangbin(
)
- Jiangxi University of Chinese Medicine Research Center for Differention and Development of Traditional Chinese Medicine (TCM) Basic Theory, Jiangxi Province Key Laboratory of TCM Etiopathogenisis, Nanchang 330004, China
-
Received:2022-07-10Accepted:2022-12-16Online:2023-04-15Published:2023-03-14 -
Contact:Prof. SHANG Guangbin, Jiangxi University of Chinese Medicine Research Center for Differention and Development of Traditional Chinese Medicine Basic Theory, Jiangxi Province Key Laboratory of TCM Etiopathogenisis, Nanchang 330004, China. shanggb@jxutcm.edu.cn. Telephone: +86-113732901725 -
Supported by:Jiangxi University of Chinese Medicine, Jiangxi Province Double First-Class Discipline of Traditional Chinese Medicine Construction Project(JXSYLXK-ZHYI026);Supported by Science and Technology Program of Jiangxi Provincial Health and Health Commission(SKJP220210415);Exploring the Formation Mechanism of Liver and Kidney Yin Deficiency Based on Animal Model and Metabolomics; Supported by National TCM Leading Talent Support Program of the State Administration of TCM-Qi Huang Scholar(284, China TCM people's teaching letter [2018])
Cite this article
HU Xingyao, LIU Hongning, YAN Xiaojun, CHEN Zhong, FU Liu, LIU Ge, CHEN Xuan, SHANG Guangbin. Liver metabolomic characteristics in three different rat models of Yin deficiency based on ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry[J]. Journal of Traditional Chinese Medicine, 2023, 43(2): 274-285.
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Table 1 Scoring criteria for appearance behavior of rats
| Observation index | Index score | ||
|---|---|---|---|
| Skin hair | Wither and easy to fall off 2 | Not shiny or fluffy 1 | Supple and shiny 0 |
| Behavioral activity | Active and frighten 2 | Hyperactivity 1 | Moderate activity 0 |
| Urinary status | Yellow and frequent urine 2 | Light yellow urine 1 | Normal urine 0 |
| Stool state | Dry stool 2 | Hard stool 1 | Normal stool 0 |
Table 1 Scoring criteria for appearance behavior of rats
| Observation index | Index score | ||
|---|---|---|---|
| Skin hair | Wither and easy to fall off 2 | Not shiny or fluffy 1 | Supple and shiny 0 |
| Behavioral activity | Active and frighten 2 | Hyperactivity 1 | Moderate activity 0 |
| Urinary status | Yellow and frequent urine 2 | Light yellow urine 1 | Normal urine 0 |
| Stool state | Dry stool 2 | Hard stool 1 | Normal stool 0 |
Table 2 Comparison of physical sign scores of rats in each group ($\bar{x}\pm s$)
| Group | n | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|
| Blank | 12 | 0.0±0.0 | 0.1±0.3 | 0.2±0.4 | 0.3±0.5 | 0.2±0.4 | 0.3±0.5 |
| Irritation induced model | 12 | 0.4±0.5 | 0.4±0.8 | 0.8±1.0 | 1.5±1.7a | 2.5±1.7b | 3.8±1.6b |
| Fuzi-Ganjiang induced model | 12 | 0.5±0.7 | 0.7±0.8 | 1.2±1.5 | 1.2±1.3 | 1.8±1.3b | 3.3±1.0b |
| Thyroxine-reserpine induced model | 12 | 0.3±0.5 | 0.4±1.0 | 1.0±1.4 | 1.3±1.3a | 2.7±1.1b | 2.5±1.1b |
Table 2 Comparison of physical sign scores of rats in each group ($\bar{x}\pm s$)
| Group | n | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|
| Blank | 12 | 0.0±0.0 | 0.1±0.3 | 0.2±0.4 | 0.3±0.5 | 0.2±0.4 | 0.3±0.5 |
| Irritation induced model | 12 | 0.4±0.5 | 0.4±0.8 | 0.8±1.0 | 1.5±1.7a | 2.5±1.7b | 3.8±1.6b |
| Fuzi-Ganjiang induced model | 12 | 0.5±0.7 | 0.7±0.8 | 1.2±1.5 | 1.2±1.3 | 1.8±1.3b | 3.3±1.0b |
| Thyroxine-reserpine induced model | 12 | 0.3±0.5 | 0.4±1.0 | 1.0±1.4 | 1.3±1.3a | 2.7±1.1b | 2.5±1.1b |
Table 3 Comparison of changes in body temperature in rats of each group (℃, $\bar{x}\pm s$)
| Group | n | Week 0 | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 36.4±0.6 | 35.8±0.7 | 36.4±0.8 | 36.4±0.5 | 36.4±0.5 | 36.0±0.6 | 36.1±0.5 |
| Irritation induced model | 12 | 36.2±1.0 | 36.2±0.8 | 36.5±0.8 | 36.7±0.6 | 37.0±0.5a | 36.7±0.7a | 36.5±0.6 |
| Fuzi-Ganjiang induced model | 12 | 36.8±1.1 | 36.2±0.6 | 35.8±0.7 | 36.4±0.6 | 36.5±0.4 | 36.7±0.5a | 36.4±0.5 |
| Thyroxine-reserpine induced model | 12 | 35.7±1.2 | 36.3±0.6 | 36.2±0.7 | 37.0±0.6a | 36.8±0.5 | 36.5±0.5b | 36.3±0.4 |
Table 3 Comparison of changes in body temperature in rats of each group (℃, $\bar{x}\pm s$)
| Group | n | Week 0 | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 36.4±0.6 | 35.8±0.7 | 36.4±0.8 | 36.4±0.5 | 36.4±0.5 | 36.0±0.6 | 36.1±0.5 |
| Irritation induced model | 12 | 36.2±1.0 | 36.2±0.8 | 36.5±0.8 | 36.7±0.6 | 37.0±0.5a | 36.7±0.7a | 36.5±0.6 |
| Fuzi-Ganjiang induced model | 12 | 36.8±1.1 | 36.2±0.6 | 35.8±0.7 | 36.4±0.6 | 36.5±0.4 | 36.7±0.5a | 36.4±0.5 |
| Thyroxine-reserpine induced model | 12 | 35.7±1.2 | 36.3±0.6 | 36.2±0.7 | 37.0±0.6a | 36.8±0.5 | 36.5±0.5b | 36.3±0.4 |
Table 4 Comparison of changes in water consumption in rats of each group (mL, $\bar{x}\pm s$)
| Group | n | Week 0 | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 33.2±3.7 | 48.1±7.7 | 54.2±4.9 | 54.2±4.9 | 61.9±9.1 | 59.2±3.1 | 61.2±7.0 |
| Irritation induced model | 12 | 33.3±4.5 | 39.7±4.6b | 49.9±5.8b | 41.3±2.9a | 56.7±8.2 | 47.2±3.2a | 53.1±6.1b |
| Fuzi-Ganjiang induced model | 12 | 31.5±4.2 | 45.6±3.0 | 49.0±3.5b | 50.0±2.0 | 59.2±4.0 | 56.5±7.5 | 52.2±4.1a |
| Thyroxine-reserpine induced model | 12 | 28.6±1.7a | 42.8±7.1 | 50.4±4.5 | 56.3±7.5 | 66.4±8.4 | 64.0±8.1 | 50.6±7.0a |
Table 4 Comparison of changes in water consumption in rats of each group (mL, $\bar{x}\pm s$)
| Group | n | Week 0 | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 33.2±3.7 | 48.1±7.7 | 54.2±4.9 | 54.2±4.9 | 61.9±9.1 | 59.2±3.1 | 61.2±7.0 |
| Irritation induced model | 12 | 33.3±4.5 | 39.7±4.6b | 49.9±5.8b | 41.3±2.9a | 56.7±8.2 | 47.2±3.2a | 53.1±6.1b |
| Fuzi-Ganjiang induced model | 12 | 31.5±4.2 | 45.6±3.0 | 49.0±3.5b | 50.0±2.0 | 59.2±4.0 | 56.5±7.5 | 52.2±4.1a |
| Thyroxine-reserpine induced model | 12 | 28.6±1.7a | 42.8±7.1 | 50.4±4.5 | 56.3±7.5 | 66.4±8.4 | 64.0±8.1 | 50.6±7.0a |
Table 5 Comparison of changes in diet in rats of each group (g, $\bar{x}\pm s$)
| Group | n | Week 0 | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 16.0±0.6 | 26.7±1.9 | 34.7±8.9 | 31.2±1.3 | 32.3±3.9 | 32.5±1.7 | 28.6±0.9 |
| Irritation induced model | 12 | 16.5±0.5 | 25.6±1.9 | 29.1±1.3 | 26.6±3.7 | 30.5±1.6 | 28.8±2.3b | 25.4±2.2b |
| Fuzi-Ganjiang induced model | 12 | 15.3±1.2 | 24.7±1.7a | 30.2±1.5 | 30.2±2.1 | 32.2±2.4 | 30.1±0.7b | 25.9±2.2b |
| Thyroxine-reserpine induced model | 12 | 13.8±1.3a | 24.8±3.3a | 34.2±2.7 | 37.3±4.7b | 41.5±1.9b | 33.5±5.1 | 31.0±1.7b |
Table 5 Comparison of changes in diet in rats of each group (g, $\bar{x}\pm s$)
| Group | n | Week 0 | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 |
|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 16.0±0.6 | 26.7±1.9 | 34.7±8.9 | 31.2±1.3 | 32.3±3.9 | 32.5±1.7 | 28.6±0.9 |
| Irritation induced model | 12 | 16.5±0.5 | 25.6±1.9 | 29.1±1.3 | 26.6±3.7 | 30.5±1.6 | 28.8±2.3b | 25.4±2.2b |
| Fuzi-Ganjiang induced model | 12 | 15.3±1.2 | 24.7±1.7a | 30.2±1.5 | 30.2±2.1 | 32.2±2.4 | 30.1±0.7b | 25.9±2.2b |
| Thyroxine-reserpine induced model | 12 | 13.8±1.3a | 24.8±3.3a | 34.2±2.7 | 37.3±4.7b | 41.5±1.9b | 33.5±5.1 | 31.0±1.7b |
Table 6 Comparison of Serum cAMP, cGMP, cAMP/cGMP, E2, T, E2/T, ALT, AST Levels in Rats of Each Group ($\bar{x}\pm s$)
| Group | n | cAMP (nmol/L) | cGMP (nmol/L) | cAMP /cGMP (nmol/L) | E2 (ng/mL) | T (ng/mL) | E2/T (ng/mL) | ALT (μ/L) | AST (μ/L) |
|---|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 39.5±8.1 | 38.7±11.4 | 0.9±0.4 | 89.2±16.3 | 9.0±5.0 | 13.6±8.8 | 32.4±8.0 | 133.7±36.3 |
| Irritation induced Model | 12 | 32.7±12.1 | 37.3±14.8 | 1.2±0.4 | 91.2±19.4 | 7.6±3.8 | 17.2±12.3 | 53.1±8.5b | 184.6±52.3b |
| Fuzi-Ganjiang induced model | 12 | 53.8±14.5a | 18.8±4.0a | 3.1±1.3a | 114.3±26.0b | 8.2±5.2 | 19.7±13.0 | 61.4±15.6a | 196.6±61.3a |
| Thyroxine-reserpine induced model | 12 | 52.1±8.8a | 15.3±3.2a | 3.5±0.8a | 127.6±30.1a | 6.7±1.5 | 26.9±18.0 | 76.0±39.0a | 209.8±54.3a |
Table 6 Comparison of Serum cAMP, cGMP, cAMP/cGMP, E2, T, E2/T, ALT, AST Levels in Rats of Each Group ($\bar{x}\pm s$)
| Group | n | cAMP (nmol/L) | cGMP (nmol/L) | cAMP /cGMP (nmol/L) | E2 (ng/mL) | T (ng/mL) | E2/T (ng/mL) | ALT (μ/L) | AST (μ/L) |
|---|---|---|---|---|---|---|---|---|---|
| Blank | 12 | 39.5±8.1 | 38.7±11.4 | 0.9±0.4 | 89.2±16.3 | 9.0±5.0 | 13.6±8.8 | 32.4±8.0 | 133.7±36.3 |
| Irritation induced Model | 12 | 32.7±12.1 | 37.3±14.8 | 1.2±0.4 | 91.2±19.4 | 7.6±3.8 | 17.2±12.3 | 53.1±8.5b | 184.6±52.3b |
| Fuzi-Ganjiang induced model | 12 | 53.8±14.5a | 18.8±4.0a | 3.1±1.3a | 114.3±26.0b | 8.2±5.2 | 19.7±13.0 | 61.4±15.6a | 196.6±61.3a |
| Thyroxine-reserpine induced model | 12 | 52.1±8.8a | 15.3±3.2a | 3.5±0.8a | 127.6±30.1a | 6.7±1.5 | 26.9±18.0 | 76.0±39.0a | 209.8±54.3a |
Table 7 Comparison of liver index and kidney index of rats in each group (%, $\bar{x}\pm s$)
| Group | Liver index | Kidney index |
|---|---|---|
| Blank | 2.58±0.09 | 0.65±0.04 |
| Irritation induced model | 2.58±0.13 | 0.66±0.03 |
| Fuzi-Ganjiang induced model | 2.89±0.37a | 0.70±0.07 |
| Thyroxine-reserpine induced model | 3.03±0.23a | 0.82±0.07a |
Table 7 Comparison of liver index and kidney index of rats in each group (%, $\bar{x}\pm s$)
| Group | Liver index | Kidney index |
|---|---|---|
| Blank | 2.58±0.09 | 0.65±0.04 |
| Irritation induced model | 2.58±0.13 | 0.66±0.03 |
| Fuzi-Ganjiang induced model | 2.89±0.37a | 0.70±0.07 |
| Thyroxine-reserpine induced model | 3.03±0.23a | 0.82±0.07a |
Figure 1 Histopathological changes of rat liver and kidney in each group (HE staining) A: blank group of liver tissue (× 100); B: blank group of liver tissue (× 400); C: irritation induced model group of liver tissue (× 100); D: irritation induced model group of liver tissue (× 400); E: Fuzi-Ganjiang induced model group of liver tissue (× 100); F: Fuzi-Ganjiang induced model group of liver tissue (× 400); G: thyroxine-reserpine induced model group of liver tissue (× 100); H: thyroxine-reserpine induced model group of liver tissue (× 400); I: blank group of kidney tissue (× 100); J: blank group of kidney tissue (× 400); K: Irritation induced model group of kidney tissue (× 100); L: irritation induced model group of kidney tissue (× 400); M: Fuzi-Ganjiang induced model group of kidney tissue (× 100); N: Fuzi-Ganjiang induced model group of kidney tissue (× 400); O: thyroxine-reserpine induced model group of kidney tissue (× 100); P: thyroxine-reserpine induced model group of kidney tissue (× 400); (Scale bar indicates 100 μm, × 100; scale bar indicates 50 μm, × 400). HE: hematoxylin-eosin.
Figure 1 Histopathological changes of rat liver and kidney in each group (HE staining) A: blank group of liver tissue (× 100); B: blank group of liver tissue (× 400); C: irritation induced model group of liver tissue (× 100); D: irritation induced model group of liver tissue (× 400); E: Fuzi-Ganjiang induced model group of liver tissue (× 100); F: Fuzi-Ganjiang induced model group of liver tissue (× 400); G: thyroxine-reserpine induced model group of liver tissue (× 100); H: thyroxine-reserpine induced model group of liver tissue (× 400); I: blank group of kidney tissue (× 100); J: blank group of kidney tissue (× 400); K: Irritation induced model group of kidney tissue (× 100); L: irritation induced model group of kidney tissue (× 400); M: Fuzi-Ganjiang induced model group of kidney tissue (× 100); N: Fuzi-Ganjiang induced model group of kidney tissue (× 400); O: thyroxine-reserpine induced model group of kidney tissue (× 100); P: thyroxine-reserpine induced model group of kidney tissue (× 400); (Scale bar indicates 100 μm, × 100; scale bar indicates 50 μm, × 400). HE: hematoxylin-eosin.
Figure 2 PCA scores of liver tissue samples of each group of animals A: positive ion mode; B: negative ion mode; 1: blank group; 2: irritation induced model group; 3: Fuzi-Ganjiang induced model group; 4: thyroxine-reserpine induced model group.
Figure 2 PCA scores of liver tissue samples of each group of animals A: positive ion mode; B: negative ion mode; 1: blank group; 2: irritation induced model group; 3: Fuzi-Ganjiang induced model group; 4: thyroxine-reserpine induced model group.
Table 8 Different endogenous compounds in rat liver
| No. | HMDB ID | Retention time(min) | m/z | Formula | Biomarkers name |
|---|---|---|---|---|---|
| 1 | HMDB0000254 | 1.047 | 117.0192 | C4H6O4 | Succinic acid |
| 2 | HMDB0006355 | 0.662 | 175.024 | C6H8O6 | D-Glucurono-6,3-lactone |
| 3 | HMDB0032387 | 4.722 | 187.0973 | C9H16O4 | (+/-)-Methyl 5-acetoxyhexanoate |
| 4 | HMDB0012204 | 1.931 | 218.1037 | C10H13N5O | Cis-zeatin |
| 5 | HMDB0000767 | 0.628 | 243.062 | C9H12N2O6 | Pseudouridine |
| 6 | HMDB0002186 | 16.312 | 253.2176 | C16H30O2 | Hypogeic acid |
| 7 | HMDB0003040 | 0.662 | 267.0727 | C10H12N4O5 | Arabinosylhypoxanthine |
| 8 | HMDB0031067 | 11.668 | 269.2483 | C17H3402 | (S)-14-Methylhexadecanoic acid |
| 9 | HMDB0010734 | 11.971 | 271.2283 | C16H32O3 | (R)-3-Hydroxy-hexadecanoic acid |
| 10 | HMDB0003426 | 2.666 | 277.1223 | C11H22N2O4S | Pantetheine |
| 11 | HMDB0001388 | 9.348 | 277.2179 | C18H30O2 | Alpha-Linolenic acid |
| 12 | HMDB0030430 | 9.916 | 279.2334 | C18H32O2 | Linalylcaprylate |
| 13 | HMDB0000207 | 17.749 | 281.2483 | C18H34O2 | Oleic acid |
| 14 | HMDB0000299 | 1.564 | 283.0683 | C10H12N4O6 | Xanthosine |
| 15 | HMDB0000827 | 19.001 | 283.2642 | C18H36O2 | Stearic acid |
| 16 | HMDB0000594 | 2.766 | 293.1146 | C14H18N2O5 | gamma-Glutamylphenylalanine |
| 17 | HMDB0037396 | 17.614 | 299.2581 | C18H36O3 | xi-10-Hydroxyoctadecanoic acid |
| 18 | HMDB0001999 | 9.234 | 301.219 | C20H30O2 | Eicosapentaenoic acid |
| 19 | HMDB0006036 | 17.614 | 303.2323 | C20H32O2 | Mesterolone |
| 20 | HMDB0002925 | 10.885 | 305.2479 | C20H34O2 | 8,11,14-Eicosatrienoic acid |
| 21 | HMDB0061650 | 12.355 | 313.2386 | C18H34O4 | 9,10-Epoxystearic acid |
| 22 | HMDB0006048 | 16.146 | 315.2337 | C21H32O2 | Bolasterone |
| 23 | HMDB0005998 | 11.318 | 319.2279 | C20H32O3 | 20-Hydroxyeicosatetraenoic acid |
| 24 | HMDB0032143 | 17.597 | 331.2637 | C22H36O2 | Palaudine |
| 25 | HMDB0035676 | 9.816 | 337.2388 | C20H34O4 | 1-Hydroxy-1-phenyl-3-hexadecanone |
| 26 | HMDB0002995 | 17.598 | 340.2854 | C20H39NO3 | 12-Keto-tetrahydro-leukotriene B4 |
| 27 | HMDB0013308 | 0.946 | 346.0556 | C10H14N5O7P | Stearoylglycine |
| 28 | HMDB0014096 | 0.678 | 350.1076 | C13H21NO10 | 5'-Hydroxypiroxicam |
| 29 | HMDB0000785 | 7.026 | 351.218 | C20H32O5 | N-Acetyl-7-O-acetylneuraminic acid |
| 30 | HMDB0035338 | 1.948 | 357.0895 | C11H23N2O7PS | Sterebin B |
| 31 | HMDB0001416 | 4.687 | 359.1258 | C18H20N2O6 | Pantetheine 4'-phosphate |
| 32 | HMDB0006045 | 14.225 | 360.2553 | C22H35NO3 | Dityrosine |
| 33 | HMDB0005096 | 16.495 | 403.1591 | C25H24O5 | N-Arachidonoyl glycine |
| 34 | HMDB0030785 | 6.676 | 423.2748 | C24H40O6 | Mammeigin |
| 35 | HMDB0013192 | 16.697 | 429.1753 | C20H30O10 | 3a,7b,21-Trihydroxy-5b-cholanoic acid |
| 36 | HMDB0032622 | 8.48 | 435.2774 | C25H40O6 | Phenethylrutinoside |
| 37 | HMDB0029949 | 18.985 | 437.2665 | C21H43O7P | Pangamic acid |
| 38 | HMDB0007850 | 6.559 | 448.3074 | C26H43NO5 | LysoPA(0:0/18:0) |
| 39 | HMDB0000708 | 9.883 | 452.2795 | C28H39NO4 | Glycoursodeoxycholic acid |
| 40 | HMDB0011473 | 5.64 | 514.285 | C26H45NO7S | LysoPE(0:0/16:0) |
| 41 | HMDB0000036 | 1.046 | 611.1452 | C20H32N6O12S2 | Taurocholic acid |
| 42 | HMDB0003337 | 19.252 | 766.5397 | C43H78NO8P | Oxidized glutathione |
| 43 | HMDB0007949 | 17.597 | 331.2637 | C22H36O2 | PC(15:0/20:4(5Z,8Z,11Z,14Z)) |
| 44 | HMDB0004226 | 0.527 | 130.0854 | C6H11NO2 | N4-Acetylaminobutanal |
| 45 | HMDB0031160 | 0.749 | 137.0453 | C5H12S2 | 1-Pentanesulfenothioic acid |
| 46 | HMDB0031210 | 3.397 | 159.0284 | C6H6O5 | Zymonic acid |
| 47 | HMDB0002052 | 5.121 | 201.0392 | C8H8O6 | Maleylacetoacetic acid |
| 48 | HMDB0011175 | 1.928 | 220.117 | C10H13N5O | Leucylproline |
| 49 | HMDB0032318 | 2.119 | 229.1538 | C11H20N2O3 | Hexanal octane-1,3-diol acetal |
| 50 | HMDB0028932 | 15.964 | 229.216 | C14H28O2 | Leucyl-Isoleucine |
| 51 | HMDB0035358 | 3.588 | 245.1858 | C12H24N2O3 | Ketosantalic acid |
| 52 | HMDB0000086 | 7.902 | 251.1637 | C15H22O3 | Glycerophosphocholine |
| 53 | HMDB0011171 | 10.05 | 258.1094 | C8H20NO6P | gamma-Glutamylleucine |
| 54 | HMDB0034495 | 2.484 | 261.1438 | C11H20N2O6 | 6,10,14-Trimethyl-5,9,13-pentadecatrien-2-one |
| 55 | HMDB0030964 | 9.916 | 263.2365 | C18H30O | Linolenelaidic acid |
| 56 | HMDB0062656 | 15.697 | 279.232 | C18H30O2 | Linoleamide |
| 57 | HMDB0061864 | 15.038 | 280.2632 | C18H33NO | Dihomolinoleic acid |
| 58 | HMDB0002117 | 16.69 | 281.2472 | C18H32O2 | Oleamide |
| 59 | HMDB0006221 | 16.478 | 282.2789 | C18H35NO | 13-cis Retinol |
| 60 | HMDB0002100 | 1.549 | 285.082 | C10H12N4O6 | Palmitoylethanolamide |
| 61 | HMDB0000269 | 19.11 | 285.2783 | C18H36O2 | Sphinganine |
| 62 | HMDB0002177 | 10.267 | 287.2366 | C20H30O | Cis-8,11,14,17-Eicosatetraenoic acid |
| 63 | HMDB0029826 | 7.425 | 300.2892 | C18H37NO2 | Hallacridone |
| 64 | HMDB0004610 | 7.225 | 318.2999 | C18H39NO3 | Phytosphingosine |
| 65 | HMDB0012252 | 7.731 | 324.2892 | C20H37NO2 | Linoleoylethanolamide |
| 66 | HMDB0002183 | 7.093 | 329.2471 | C22H32O2 | Docosahexaenoic acid |
| 67 | HMDB0032476 | 19.304 | 341.3041 | C21H40O3 | Polyoxyethylene (600) monoricinoleate |
| No. | HMDB ID | Retention time(min) | m/z | Formula | Biomarkers Name |
| 68 | HMDB0002007 | 9.795 | 357.2784 | C24H36O2 | Tetracosahexaenoic acid |
| 69 | HMDB0034031 | 16.595 | 357.3002 | C21H40O4 | 3-(2-Heptenyloxy)-2-hydroxypropyl undecanoate |
| 70 | HMDB0000476 | 14.22 | 362.2687 | C22H35NO3 | 3-Oxo-4,6-choladienoic acid |
| 71 | HMDB0013627 | 6.838 | 371.2574 | C24H34O3 | Cervonoylethanolamide |
| 72 | HMDB0012866 | 8.513 | 373.2734 | C24H36O3 | 9'-Carboxy-alpha-chromanol |
| 73 | HMDB0006898 | 8.494 | 391.2835 | C24H38O4 | Chenodeoxyglycocholic acid |
| 74 | HMDB0000331 | 8.265 | 450.3206 | C26H43NO5 | 3a,7b,12a-Trihydroxyoxocholanyl-Glycine |
| 75 | HMDB0011475 | 5.672 | 466.3154 | C26H43NO6 | LysoPE (0:0/18:1(11Z)) |
| 76 | HMDB0011129 | 11.104 | 480.3083 | C23H46NO7P | LysoPE (0:0/18:0) |
| 77 | HMDB0010395 | 9.73 | 482.3234 | C23H48NO7P | LysoPC (20:4(5Z,8Z,11Z,14Z)) |
| 78 | HMDB0008177 | 6.612 | 516.2981 | C26H45NO7P | PC (18:3(6Z,9Z,12Z)/20:2(11Z,14Z)) |
Table 8 Different endogenous compounds in rat liver
| No. | HMDB ID | Retention time(min) | m/z | Formula | Biomarkers name |
|---|---|---|---|---|---|
| 1 | HMDB0000254 | 1.047 | 117.0192 | C4H6O4 | Succinic acid |
| 2 | HMDB0006355 | 0.662 | 175.024 | C6H8O6 | D-Glucurono-6,3-lactone |
| 3 | HMDB0032387 | 4.722 | 187.0973 | C9H16O4 | (+/-)-Methyl 5-acetoxyhexanoate |
| 4 | HMDB0012204 | 1.931 | 218.1037 | C10H13N5O | Cis-zeatin |
| 5 | HMDB0000767 | 0.628 | 243.062 | C9H12N2O6 | Pseudouridine |
| 6 | HMDB0002186 | 16.312 | 253.2176 | C16H30O2 | Hypogeic acid |
| 7 | HMDB0003040 | 0.662 | 267.0727 | C10H12N4O5 | Arabinosylhypoxanthine |
| 8 | HMDB0031067 | 11.668 | 269.2483 | C17H3402 | (S)-14-Methylhexadecanoic acid |
| 9 | HMDB0010734 | 11.971 | 271.2283 | C16H32O3 | (R)-3-Hydroxy-hexadecanoic acid |
| 10 | HMDB0003426 | 2.666 | 277.1223 | C11H22N2O4S | Pantetheine |
| 11 | HMDB0001388 | 9.348 | 277.2179 | C18H30O2 | Alpha-Linolenic acid |
| 12 | HMDB0030430 | 9.916 | 279.2334 | C18H32O2 | Linalylcaprylate |
| 13 | HMDB0000207 | 17.749 | 281.2483 | C18H34O2 | Oleic acid |
| 14 | HMDB0000299 | 1.564 | 283.0683 | C10H12N4O6 | Xanthosine |
| 15 | HMDB0000827 | 19.001 | 283.2642 | C18H36O2 | Stearic acid |
| 16 | HMDB0000594 | 2.766 | 293.1146 | C14H18N2O5 | gamma-Glutamylphenylalanine |
| 17 | HMDB0037396 | 17.614 | 299.2581 | C18H36O3 | xi-10-Hydroxyoctadecanoic acid |
| 18 | HMDB0001999 | 9.234 | 301.219 | C20H30O2 | Eicosapentaenoic acid |
| 19 | HMDB0006036 | 17.614 | 303.2323 | C20H32O2 | Mesterolone |
| 20 | HMDB0002925 | 10.885 | 305.2479 | C20H34O2 | 8,11,14-Eicosatrienoic acid |
| 21 | HMDB0061650 | 12.355 | 313.2386 | C18H34O4 | 9,10-Epoxystearic acid |
| 22 | HMDB0006048 | 16.146 | 315.2337 | C21H32O2 | Bolasterone |
| 23 | HMDB0005998 | 11.318 | 319.2279 | C20H32O3 | 20-Hydroxyeicosatetraenoic acid |
| 24 | HMDB0032143 | 17.597 | 331.2637 | C22H36O2 | Palaudine |
| 25 | HMDB0035676 | 9.816 | 337.2388 | C20H34O4 | 1-Hydroxy-1-phenyl-3-hexadecanone |
| 26 | HMDB0002995 | 17.598 | 340.2854 | C20H39NO3 | 12-Keto-tetrahydro-leukotriene B4 |
| 27 | HMDB0013308 | 0.946 | 346.0556 | C10H14N5O7P | Stearoylglycine |
| 28 | HMDB0014096 | 0.678 | 350.1076 | C13H21NO10 | 5'-Hydroxypiroxicam |
| 29 | HMDB0000785 | 7.026 | 351.218 | C20H32O5 | N-Acetyl-7-O-acetylneuraminic acid |
| 30 | HMDB0035338 | 1.948 | 357.0895 | C11H23N2O7PS | Sterebin B |
| 31 | HMDB0001416 | 4.687 | 359.1258 | C18H20N2O6 | Pantetheine 4'-phosphate |
| 32 | HMDB0006045 | 14.225 | 360.2553 | C22H35NO3 | Dityrosine |
| 33 | HMDB0005096 | 16.495 | 403.1591 | C25H24O5 | N-Arachidonoyl glycine |
| 34 | HMDB0030785 | 6.676 | 423.2748 | C24H40O6 | Mammeigin |
| 35 | HMDB0013192 | 16.697 | 429.1753 | C20H30O10 | 3a,7b,21-Trihydroxy-5b-cholanoic acid |
| 36 | HMDB0032622 | 8.48 | 435.2774 | C25H40O6 | Phenethylrutinoside |
| 37 | HMDB0029949 | 18.985 | 437.2665 | C21H43O7P | Pangamic acid |
| 38 | HMDB0007850 | 6.559 | 448.3074 | C26H43NO5 | LysoPA(0:0/18:0) |
| 39 | HMDB0000708 | 9.883 | 452.2795 | C28H39NO4 | Glycoursodeoxycholic acid |
| 40 | HMDB0011473 | 5.64 | 514.285 | C26H45NO7S | LysoPE(0:0/16:0) |
| 41 | HMDB0000036 | 1.046 | 611.1452 | C20H32N6O12S2 | Taurocholic acid |
| 42 | HMDB0003337 | 19.252 | 766.5397 | C43H78NO8P | Oxidized glutathione |
| 43 | HMDB0007949 | 17.597 | 331.2637 | C22H36O2 | PC(15:0/20:4(5Z,8Z,11Z,14Z)) |
| 44 | HMDB0004226 | 0.527 | 130.0854 | C6H11NO2 | N4-Acetylaminobutanal |
| 45 | HMDB0031160 | 0.749 | 137.0453 | C5H12S2 | 1-Pentanesulfenothioic acid |
| 46 | HMDB0031210 | 3.397 | 159.0284 | C6H6O5 | Zymonic acid |
| 47 | HMDB0002052 | 5.121 | 201.0392 | C8H8O6 | Maleylacetoacetic acid |
| 48 | HMDB0011175 | 1.928 | 220.117 | C10H13N5O | Leucylproline |
| 49 | HMDB0032318 | 2.119 | 229.1538 | C11H20N2O3 | Hexanal octane-1,3-diol acetal |
| 50 | HMDB0028932 | 15.964 | 229.216 | C14H28O2 | Leucyl-Isoleucine |
| 51 | HMDB0035358 | 3.588 | 245.1858 | C12H24N2O3 | Ketosantalic acid |
| 52 | HMDB0000086 | 7.902 | 251.1637 | C15H22O3 | Glycerophosphocholine |
| 53 | HMDB0011171 | 10.05 | 258.1094 | C8H20NO6P | gamma-Glutamylleucine |
| 54 | HMDB0034495 | 2.484 | 261.1438 | C11H20N2O6 | 6,10,14-Trimethyl-5,9,13-pentadecatrien-2-one |
| 55 | HMDB0030964 | 9.916 | 263.2365 | C18H30O | Linolenelaidic acid |
| 56 | HMDB0062656 | 15.697 | 279.232 | C18H30O2 | Linoleamide |
| 57 | HMDB0061864 | 15.038 | 280.2632 | C18H33NO | Dihomolinoleic acid |
| 58 | HMDB0002117 | 16.69 | 281.2472 | C18H32O2 | Oleamide |
| 59 | HMDB0006221 | 16.478 | 282.2789 | C18H35NO | 13-cis Retinol |
| 60 | HMDB0002100 | 1.549 | 285.082 | C10H12N4O6 | Palmitoylethanolamide |
| 61 | HMDB0000269 | 19.11 | 285.2783 | C18H36O2 | Sphinganine |
| 62 | HMDB0002177 | 10.267 | 287.2366 | C20H30O | Cis-8,11,14,17-Eicosatetraenoic acid |
| 63 | HMDB0029826 | 7.425 | 300.2892 | C18H37NO2 | Hallacridone |
| 64 | HMDB0004610 | 7.225 | 318.2999 | C18H39NO3 | Phytosphingosine |
| 65 | HMDB0012252 | 7.731 | 324.2892 | C20H37NO2 | Linoleoylethanolamide |
| 66 | HMDB0002183 | 7.093 | 329.2471 | C22H32O2 | Docosahexaenoic acid |
| 67 | HMDB0032476 | 19.304 | 341.3041 | C21H40O3 | Polyoxyethylene (600) monoricinoleate |
| No. | HMDB ID | Retention time(min) | m/z | Formula | Biomarkers Name |
| 68 | HMDB0002007 | 9.795 | 357.2784 | C24H36O2 | Tetracosahexaenoic acid |
| 69 | HMDB0034031 | 16.595 | 357.3002 | C21H40O4 | 3-(2-Heptenyloxy)-2-hydroxypropyl undecanoate |
| 70 | HMDB0000476 | 14.22 | 362.2687 | C22H35NO3 | 3-Oxo-4,6-choladienoic acid |
| 71 | HMDB0013627 | 6.838 | 371.2574 | C24H34O3 | Cervonoylethanolamide |
| 72 | HMDB0012866 | 8.513 | 373.2734 | C24H36O3 | 9'-Carboxy-alpha-chromanol |
| 73 | HMDB0006898 | 8.494 | 391.2835 | C24H38O4 | Chenodeoxyglycocholic acid |
| 74 | HMDB0000331 | 8.265 | 450.3206 | C26H43NO5 | 3a,7b,12a-Trihydroxyoxocholanyl-Glycine |
| 75 | HMDB0011475 | 5.672 | 466.3154 | C26H43NO6 | LysoPE (0:0/18:1(11Z)) |
| 76 | HMDB0011129 | 11.104 | 480.3083 | C23H46NO7P | LysoPE (0:0/18:0) |
| 77 | HMDB0010395 | 9.73 | 482.3234 | C23H48NO7P | LysoPC (20:4(5Z,8Z,11Z,14Z)) |
| 78 | HMDB0008177 | 6.612 | 516.2981 | C26H45NO7P | PC (18:3(6Z,9Z,12Z)/20:2(11Z,14Z)) |
Table 9 Analysis of key metabolic pathways in rat liver tissue metabolism
| No. | Pathway Name | Total | Hits | Raw p | Holm adjust | FDR p | Impact |
|---|---|---|---|---|---|---|---|
| 1 | Biosynthesis of unsaturated fatty acids | 36 | 6 | 0.000099 | 0.008348 | 0.008348 | 0 |
| 2 | Glycerophospholipid metabolism | 36 | 4 | 0.0076 | 0.63076 | 0.31918 | 0.29983 |
| 3 | alpha-Linolenic acid metabolism | 13 | 2 | 0.032911 | 1 | 0.92151 | 0.33333 |
| 4 | Arachidonic acid metabolism | 36 | 3 | 0.044951 | 1 | 0.94396 | 0.0212 |
| 5 | Pantothenate and CoA biosynthesis | 19 | 2 | 0.06634 | 1 | 1 | 0.27857 |
| 6 | Sphingolipid metabolism | 21 | 2 | 0.079235 | 1 | 1 | 0.15822 |
| 7 | Linoleic acid metabolism | 5 | 1 | 0.10783 | 1 | 1 | 0 |
| 8 | Taurine and hypotaurine metabolism | 8 | 1 | 0.16702 | 1 | 1 | 0 |
| 9 | Ascorbate and aldarate metabolism | 10 | 1 | 0.20434 | 1 | 1 | 0 |
| 10 | Butanoate metabolism | 15 | 1 | 0.29068 | 1 | 1 | 0 |
| 11 | Glycerolipid metabolism | 16 | 1 | 0.30683 | 1 | 1 | 0.01246 |
| 12 | Ether lipid metabolism | 20 | 1 | 0.3679 | 1 | 1 | 0 |
| 13 | Citrate cycle (TCA cycle) | 20 | 1 | 0.3679 | 1 | 1 | 0.03273 |
| 14 | Propanoate metabolism | 23 | 1 | 0.41025 | 1 | 1 | 0 |
| 15 | Alanine, aspartate and glutamate metabolism | 28 | 1 | 0.47478 | 1 | 1 | 0 |
| 16 | Phosphatidylinositol signaling system | 28 | 1 | 0.47478 | 1 | 1 | 0.00152 |
| 17 | Glutathione metabolism | 28 | 1 | 0.47478 | 1 | 1 | 0.02698 |
| 18 | Arginine and proline metabolism | 38 | 1 | 0.58393 | 1 | 1 | 0.01212 |
| 19 | Tyrosine metabolism | 42 | 1 | 0.62112 | 1 | 1 | 0.05581 |
| 20 | Primary bile acid biosynthesis | 46 | 1 | 0.65508 | 1 | 1 | 0.02285 |
| 21 | Purine metabolism | 66 | 1 | 0.78516 | 1 | 1 | 0 |
Table 9 Analysis of key metabolic pathways in rat liver tissue metabolism
| No. | Pathway Name | Total | Hits | Raw p | Holm adjust | FDR p | Impact |
|---|---|---|---|---|---|---|---|
| 1 | Biosynthesis of unsaturated fatty acids | 36 | 6 | 0.000099 | 0.008348 | 0.008348 | 0 |
| 2 | Glycerophospholipid metabolism | 36 | 4 | 0.0076 | 0.63076 | 0.31918 | 0.29983 |
| 3 | alpha-Linolenic acid metabolism | 13 | 2 | 0.032911 | 1 | 0.92151 | 0.33333 |
| 4 | Arachidonic acid metabolism | 36 | 3 | 0.044951 | 1 | 0.94396 | 0.0212 |
| 5 | Pantothenate and CoA biosynthesis | 19 | 2 | 0.06634 | 1 | 1 | 0.27857 |
| 6 | Sphingolipid metabolism | 21 | 2 | 0.079235 | 1 | 1 | 0.15822 |
| 7 | Linoleic acid metabolism | 5 | 1 | 0.10783 | 1 | 1 | 0 |
| 8 | Taurine and hypotaurine metabolism | 8 | 1 | 0.16702 | 1 | 1 | 0 |
| 9 | Ascorbate and aldarate metabolism | 10 | 1 | 0.20434 | 1 | 1 | 0 |
| 10 | Butanoate metabolism | 15 | 1 | 0.29068 | 1 | 1 | 0 |
| 11 | Glycerolipid metabolism | 16 | 1 | 0.30683 | 1 | 1 | 0.01246 |
| 12 | Ether lipid metabolism | 20 | 1 | 0.3679 | 1 | 1 | 0 |
| 13 | Citrate cycle (TCA cycle) | 20 | 1 | 0.3679 | 1 | 1 | 0.03273 |
| 14 | Propanoate metabolism | 23 | 1 | 0.41025 | 1 | 1 | 0 |
| 15 | Alanine, aspartate and glutamate metabolism | 28 | 1 | 0.47478 | 1 | 1 | 0 |
| 16 | Phosphatidylinositol signaling system | 28 | 1 | 0.47478 | 1 | 1 | 0.00152 |
| 17 | Glutathione metabolism | 28 | 1 | 0.47478 | 1 | 1 | 0.02698 |
| 18 | Arginine and proline metabolism | 38 | 1 | 0.58393 | 1 | 1 | 0.01212 |
| 19 | Tyrosine metabolism | 42 | 1 | 0.62112 | 1 | 1 | 0.05581 |
| 20 | Primary bile acid biosynthesis | 46 | 1 | 0.65508 | 1 | 1 | 0.02285 |
| 21 | Purine metabolism | 66 | 1 | 0.78516 | 1 | 1 | 0 |
Figure 3 Enrichment diagram of key metabolic pathways in rats liver metabolism with Yin deficiency
Figure 3 Enrichment diagram of key metabolic pathways in rats liver metabolism with Yin deficiency
Figure 4 Metabolism diagram of liver tissues in rats with Yin deficiency A: biosynthesis of pantothenic acid and CoA; B: α-linolenic acid metabolism; C: glycero-phospholipid metabolism; D: sphingolipid metabolism.
Figure 4 Metabolism diagram of liver tissues in rats with Yin deficiency A: biosynthesis of pantothenic acid and CoA; B: α-linolenic acid metabolism; C: glycero-phospholipid metabolism; D: sphingolipid metabolism.
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