Journal of Traditional Chinese Medicine ›› 2025, Vol. 45 ›› Issue (5): 1009-1018.DOI: 10.19852/j.cnki.jtcm.20250327.001
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Liquid chromatography-mass spectrometry-based metabolomics study of the protective mechanism of Shenji Guben decoction (参吉固本方) on chronic obstructive pulmonary disease
PAN Tingyu1, YAO Jing1,2, GE Yue1, YANG Shuang1, SUN Zikai1, WEI Yu1, WU Jieyu1, XU Yong1,3, ZHOU Xianmei1(
), HE Hailang1()
- 1 Department of Pulmonary and Critical Care Medicine, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210018, China
2 Department of Pulmonary Medicine, Suzhou Hospital of Integrated Traditional Chinese and Western Medicine, Suzhou 215101, China
3 School of Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China
-
Received:2024-09-11Accepted:2024-12-22Online:2025-10-15Published:2025-03-27 -
Contact:HE Hailang, Department of Pulmonary and Critical Care Medicine, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210018, China. lyghehailang@163.com;
Prof. ZHOU Xianmei, Department of Pulmonary and Critical Care Medicine, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210018, China. zhouxianmeijs@aliyun.com, Telephone: +86-25-86617141; +86-15950585516 -
About author:PAN Tingyu and YAO Jing are co-first authors and contributed equally to this work -
Supported by:Traditional Chinese Medicine Science and Technology Development Plan of Jiangsu Province(MS2021013);Traditional Chinese Medicine Science and Technology Development Plan of Jiangsu Province(ZD202215);Exploring the Efficacy and Mechanism of Professor Cao Shihong's Shenji Guben Decoction in Treating Chronic Obstructive Pulmonary Disease Based on Metabolomics, a Real-World Cohort Study on Traditional Chinese Medicine for the Treatment of Acute Exacerbation of Chronic Obstructive Pulmonary Disease;Intra-hospital Fund of Jiangsu Provincial Hospital of Traditional Chinese Medicine Development(Y2021ZR11);Investigating the Mechanism of Shenji Guben Decoction in Treating Chronic Obstructive Pulmonary Disease Based on Metabolomics
Cite this article
PAN Tingyu, YAO Jing, GE Yue, YANG Shuang, SUN Zikai, WEI Yu, WU Jieyu, XU Yong, ZHOU Xianmei, HE Hailang. Liquid chromatography-mass spectrometry-based metabolomics study of the protective mechanism of Shenji Guben decoction (参吉固本方) on chronic obstructive pulmonary disease[J]. Journal of Traditional Chinese Medicine, 2025, 45(5): 1009-1018.
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Figure 1 Identification of active compounds in SJGB decoction using UHPLC-MS/MS A: SJGB decoction in ESI+ mode; B: SJGB decoction in ESI-mode. SJGB: Shenji Guben; UHPLC-MS/MS: ultra-high performance liquid chromatography-tandem mass spectrometry; ESI: electrospray ionization.
Figure 1 Identification of active compounds in SJGB decoction using UHPLC-MS/MS A: SJGB decoction in ESI+ mode; B: SJGB decoction in ESI-mode. SJGB: Shenji Guben; UHPLC-MS/MS: ultra-high performance liquid chromatography-tandem mass spectrometry; ESI: electrospray ionization.
Table 1 Chemical characterization of bioactive compounds in SJGB decoction
| Number | RT | Identification | m/z | Formula | Class |
|---|---|---|---|---|---|
| 1 | 6.231 | (S)-Actinidine | 148.11188 | C10H13N | Pyridines and derivatives |
| 2 | 6.18 | Ephedrine | 166.12236 | C10H15NO | Benzene and substituted derivatives |
| 3 | 2.444 | Pyroglutamic acid | 188.05511 | C5H7NO3 | Carboxylic acids and derivatives |
| 4 | 6.575 | Methylephedrine | 180.13794 | C11H17NO | Benzene and substituted derivatives |
| 5 | 11.957 | Coumarin | 147.04408 | C9H6O2 | Coumarins and derivatives |
| 6 | 4.221 | 3-Hexenedioic acid | 145.04926 | C6H8O4 | Fatty Acyls |
| 7 | 11.52 | Salvianolic Acid B | 717.14667 | C36H30O16 | 2-arylbenzofuran flavonoids |
| 8 | 5.542 | Adenosine | 266.08945 | C10H13N5O4 | Purine nucleosides |
| 9 | 1.754 | Adenine | 134.04568 | C5H5N5 | Imidazopyrimidines |
| 10 | 1.197 | Choline | 104.10737 | C5H14NO | Organonitrogen compounds |
| 11 | 2.337 | Isoleucine | 132.10184 | C6H13NO2 | Carboxylic acids and derivatives |
| 12 | 1.836 | Citric acid | 191.01862 | C6H8O7 | Carboxylic acids and derivatives |
| 13 | 15.513 | 2-Methoxycinnamaldehyde | 163.07513 | C10H10O2 | Cinnamaldehydes |
| 14 | 6.537 | Maltol | 127.03901 | C6H6O3 | Pyrans |
| 15 | 20.769 | Cryptotanshinone | 297.14835 | C19H20O3 | Prenol lipids |
| 16 | 2.707 | Tyrosine | 182.08083 | C9H11NO3 | Carboxylic acids and derivatives |
| 17 | 5.793 | Phenylalanine | 166.0863 | C9H11NO2 | Carboxylic acids and derivatives |
| 18 | 9.618 | Isoquercitrin | 463.08783 | C21H20O12 | Flavonoids |
| 19 | 5.793 | Phenylethanolamine | 120.08096 | C8H11NO | Organonitrogen compounds |
| 20 | 1.208 | Malic acid | 133.01283 | C4H6O5 | Hydroxy acids and derivatives |
| 21 | 0.849 | Arginine | 175.11867 | C6H14N4O2 | Carboxylic acids and derivatives |
| 22 | 8.666 | Abrine | 241.09697 | C12H14N2O2 | Carboxylic acids and derivatives |
| 23 | 12.962 | Calycosin | 285.0755 | C16H12O5 | Isoflavonoids |
| 24 | 0.958 | Proline | 116.07094 | C5H9NO2 | Carboxylic acids and derivatives |
| 25 | 10.903 | Rosmarinic acid | 359.07648 | C18H16O8 | Cinnamic acids and derivatives |
| 26 | 10.47 | Astragalin | 447.09238 | C21H20O11 | Flavonoids |
| 27 | 11.545 | Ononin | 475.12485 | C22H22O9 | Isoflavonoids |
| 28 | 2.131 | Nicotinamide | 123.05542 | C6H6N2O | Pyridines and derivatives |
| 29 | 1.107 | Sucrose | 341.10867 | C12H22O11 | Organooxygen compounds |
| 30 | 8.906 | Glycitin | 447.12900 | C22H22O10 | Isoflavonoids |
Table 1 Chemical characterization of bioactive compounds in SJGB decoction
| Number | RT | Identification | m/z | Formula | Class |
|---|---|---|---|---|---|
| 1 | 6.231 | (S)-Actinidine | 148.11188 | C10H13N | Pyridines and derivatives |
| 2 | 6.18 | Ephedrine | 166.12236 | C10H15NO | Benzene and substituted derivatives |
| 3 | 2.444 | Pyroglutamic acid | 188.05511 | C5H7NO3 | Carboxylic acids and derivatives |
| 4 | 6.575 | Methylephedrine | 180.13794 | C11H17NO | Benzene and substituted derivatives |
| 5 | 11.957 | Coumarin | 147.04408 | C9H6O2 | Coumarins and derivatives |
| 6 | 4.221 | 3-Hexenedioic acid | 145.04926 | C6H8O4 | Fatty Acyls |
| 7 | 11.52 | Salvianolic Acid B | 717.14667 | C36H30O16 | 2-arylbenzofuran flavonoids |
| 8 | 5.542 | Adenosine | 266.08945 | C10H13N5O4 | Purine nucleosides |
| 9 | 1.754 | Adenine | 134.04568 | C5H5N5 | Imidazopyrimidines |
| 10 | 1.197 | Choline | 104.10737 | C5H14NO | Organonitrogen compounds |
| 11 | 2.337 | Isoleucine | 132.10184 | C6H13NO2 | Carboxylic acids and derivatives |
| 12 | 1.836 | Citric acid | 191.01862 | C6H8O7 | Carboxylic acids and derivatives |
| 13 | 15.513 | 2-Methoxycinnamaldehyde | 163.07513 | C10H10O2 | Cinnamaldehydes |
| 14 | 6.537 | Maltol | 127.03901 | C6H6O3 | Pyrans |
| 15 | 20.769 | Cryptotanshinone | 297.14835 | C19H20O3 | Prenol lipids |
| 16 | 2.707 | Tyrosine | 182.08083 | C9H11NO3 | Carboxylic acids and derivatives |
| 17 | 5.793 | Phenylalanine | 166.0863 | C9H11NO2 | Carboxylic acids and derivatives |
| 18 | 9.618 | Isoquercitrin | 463.08783 | C21H20O12 | Flavonoids |
| 19 | 5.793 | Phenylethanolamine | 120.08096 | C8H11NO | Organonitrogen compounds |
| 20 | 1.208 | Malic acid | 133.01283 | C4H6O5 | Hydroxy acids and derivatives |
| 21 | 0.849 | Arginine | 175.11867 | C6H14N4O2 | Carboxylic acids and derivatives |
| 22 | 8.666 | Abrine | 241.09697 | C12H14N2O2 | Carboxylic acids and derivatives |
| 23 | 12.962 | Calycosin | 285.0755 | C16H12O5 | Isoflavonoids |
| 24 | 0.958 | Proline | 116.07094 | C5H9NO2 | Carboxylic acids and derivatives |
| 25 | 10.903 | Rosmarinic acid | 359.07648 | C18H16O8 | Cinnamic acids and derivatives |
| 26 | 10.47 | Astragalin | 447.09238 | C21H20O11 | Flavonoids |
| 27 | 11.545 | Ononin | 475.12485 | C22H22O9 | Isoflavonoids |
| 28 | 2.131 | Nicotinamide | 123.05542 | C6H6N2O | Pyridines and derivatives |
| 29 | 1.107 | Sucrose | 341.10867 | C12H22O11 | Organooxygen compounds |
| 30 | 8.906 | Glycitin | 447.12900 | C22H22O10 | Isoflavonoids |
Figure 2 SJGB decoction alleviated COPD in model mice A: changes in body weight of mice in different groups; B: final body weight of mice; C: the lung function of control and model mice at the end of week 12; D: the lung function of mice at the end of week 16 stimulated with 50 mg/mL acetylcholine. SJGB-l: low dose group (7.5 g·kg-1·d-1) once daily; SJGB-m: medium dose group (15 g·kg-1·d-1) once daily; SJGB-h: high dose group (30 g·kg-1·d-1) once daily. NAC: positive drug group (78 mg·kg-1·d-1) twice daily; the model and control groups received distilled water once daily. SJGB: Shenji Guben; NAC: N-Acetylcysteine; Penh: enhanced pause; COPD: chronic obstructive pulmonary disease. The data were expressed as mean ± standard deviation and analysed using one-way analysis of variance with Tukey’s post hoc test. For Figures A and B, n = 10; for Figures C and D, n = 6. aP < 0.01 model group vs normal group, bP < 0.01 vs model group, cP < 0.05 vs model group.
Figure 2 SJGB decoction alleviated COPD in model mice A: changes in body weight of mice in different groups; B: final body weight of mice; C: the lung function of control and model mice at the end of week 12; D: the lung function of mice at the end of week 16 stimulated with 50 mg/mL acetylcholine. SJGB-l: low dose group (7.5 g·kg-1·d-1) once daily; SJGB-m: medium dose group (15 g·kg-1·d-1) once daily; SJGB-h: high dose group (30 g·kg-1·d-1) once daily. NAC: positive drug group (78 mg·kg-1·d-1) twice daily; the model and control groups received distilled water once daily. SJGB: Shenji Guben; NAC: N-Acetylcysteine; Penh: enhanced pause; COPD: chronic obstructive pulmonary disease. The data were expressed as mean ± standard deviation and analysed using one-way analysis of variance with Tukey’s post hoc test. For Figures A and B, n = 10; for Figures C and D, n = 6. aP < 0.01 model group vs normal group, bP < 0.01 vs model group, cP < 0.05 vs model group.
Figure 3 SJGB decoction improves lung tissue pathology in COPD model A1-A6: HE staining of lung tissue histological sections in each group at the end of week 16 (×100); A7-A12: HE staining of the perialveolar area (×200); A13-A18: HE staining of the peribronchial area (× 200); A1, A7, A13: Control group; A2, A8, A14: Model group; A3, A9, A15: SJGB-l group; A4, A10, A16: SJGB-m group; A5, A11, A17: SJGB-h group; A6, A12, A18: NAC group; B: peribronchial inflammation score; C: MLI. SJGB-l: low dose group (7.5 g·kg-1·d-1) once daily; SJGB-m: medium dose group (15 g·kg-1·d-1) once daily; SJGB-h: high dose group (30 g·kg-1·d-1) once daily. NAC: positive drug group (78 mg·kg-1·d-1) twice daily; the model and control groups received distilled water once daily. HE: hematoxylin-eosin; SJGB: Shenji Guben; NAC: N-Acetylcysteine; MLI: mean linear intercept; COPD: chronic obstructive pulmonary disease. The data were expressed as mean ± standard deviation (n = 3) and analysed using one-way analysis of variance with Tukey’s post hoc test. aP < 0.01 model group vs normal group; bP < 0.05 vs model group; cP < 0.01 vs model group.
Figure 3 SJGB decoction improves lung tissue pathology in COPD model A1-A6: HE staining of lung tissue histological sections in each group at the end of week 16 (×100); A7-A12: HE staining of the perialveolar area (×200); A13-A18: HE staining of the peribronchial area (× 200); A1, A7, A13: Control group; A2, A8, A14: Model group; A3, A9, A15: SJGB-l group; A4, A10, A16: SJGB-m group; A5, A11, A17: SJGB-h group; A6, A12, A18: NAC group; B: peribronchial inflammation score; C: MLI. SJGB-l: low dose group (7.5 g·kg-1·d-1) once daily; SJGB-m: medium dose group (15 g·kg-1·d-1) once daily; SJGB-h: high dose group (30 g·kg-1·d-1) once daily. NAC: positive drug group (78 mg·kg-1·d-1) twice daily; the model and control groups received distilled water once daily. HE: hematoxylin-eosin; SJGB: Shenji Guben; NAC: N-Acetylcysteine; MLI: mean linear intercept; COPD: chronic obstructive pulmonary disease. The data were expressed as mean ± standard deviation (n = 3) and analysed using one-way analysis of variance with Tukey’s post hoc test. aP < 0.01 model group vs normal group; bP < 0.05 vs model group; cP < 0.01 vs model group.
Figure 4 Metabolomics analysis A: PLS-DA score plot; B1: pairwise comparisons of Prostaglandin E2; B2: pairwise comparisons of DL-Citrulline; C: volcano plot between the control group and COPD model group; D: volcano plot between the SJGB decoction-treated group and model group; E: pathway analysis overview depicting the altered metabolic pathways in the SJGB decoction-treated group and model group. The model and control groups: distilled water once daily. SJGB-h: high dose group (30 g·kg-1·d-1) once daily. SJGB: Shenji Guben; Conc: concentration; PC: phosphatidylcholine; PE: phosphatidylethanolamine; PS: phosphatidylserine; SM: sphingomyelin; FC: fold change; PLS-DA: partial least squares discriminant analysis; COPD: chronic obstructive pulmonary disease. The data were expressed as mean ± standard deviation (n = 6) and intergroup comparison was performed using Student’s t-test. aP < 0.05 model group vs normal group; bP < 0.01 vs model group.
Figure 4 Metabolomics analysis A: PLS-DA score plot; B1: pairwise comparisons of Prostaglandin E2; B2: pairwise comparisons of DL-Citrulline; C: volcano plot between the control group and COPD model group; D: volcano plot between the SJGB decoction-treated group and model group; E: pathway analysis overview depicting the altered metabolic pathways in the SJGB decoction-treated group and model group. The model and control groups: distilled water once daily. SJGB-h: high dose group (30 g·kg-1·d-1) once daily. SJGB: Shenji Guben; Conc: concentration; PC: phosphatidylcholine; PE: phosphatidylethanolamine; PS: phosphatidylserine; SM: sphingomyelin; FC: fold change; PLS-DA: partial least squares discriminant analysis; COPD: chronic obstructive pulmonary disease. The data were expressed as mean ± standard deviation (n = 6) and intergroup comparison was performed using Student’s t-test. aP < 0.05 model group vs normal group; bP < 0.01 vs model group.
Table 2 Disturbed metabolites with P < 0.05 in the high-dose SJGB decoction-treated group compared to the model group
| Metabolite | HMDB ID/ PubChem CIDa | Fold change SJGB vs M | P value SJGB vs M |
|---|---|---|---|
| PC (18:0/18:2(9Z,12Z)) | HMDB0008039 | 0.664 | 0.001 |
| PC (18:2(9Z,12Z)/20:3(5Z,8Z,11Z)) | HMDB0008139 | 0.669 | 0.007 |
| PC (38:4) | HMDB0007988 | 0.671 | 0.024 |
| PS (15:0/24:1(15Z)) | HMDB0112341 | 0.677 | 0.015 |
| PC (18:0/20:4(8Z,11Z,14Z,17Z)) | HMDB0008049 | 0.693 | 0.026 |
| PE-NMe (20:4(8Z,11Z,14Z,17Z)/20:0) | HMDB0113462 | 0.708 | 0.037 |
| PC (36:4) | HMDB0007889 | 0.763 | 0.041 |
| PC (34:1) | HMDB0007879 | 0.803 | 0.047 |
| Thioetheramide-pc | PubChem CID: 10259406 | 0.828 | 0.020 |
| Galactosylhydroxylysine | HMDB0000600 | 0.843 | 0.046 |
| 2-Hydroxyadipic acid | HMDB0000321 | 0.862 | 0.000 |
| Floionolic acid | HMDB0034295 | 1.064 | 0.034 |
| 4-Nitrophenol | HMDB0001232 | 1.077 | 0.039 |
| 5b-Cholestane-3a,7a,12a,23R,25-pentol | HMDB0000513 | 1.153 | 0.021 |
| Glycocholic Acid | HMDB0000331 | 1.195 | 0.010 |
| 2-Hydroxy-6-Pentadecylbenzoic Acid | HMDB0029683 | 1.211 | 0.023 |
| Astromicin | HMDB0248678 | 1.223 | 0.049 |
| N-Glycolylneuraminic acid | HMDB0000833 | 1.228 | 0.040 |
| 5'-Adenylic acid | HMDB0000045 | 1.252 | 0.020 |
| Riboflavin | HMDB0000244 | 1.267 | 0.025 |
| LysoPI (20:4(5Z,8Z,11Z,14Z)/0:0) | HMDB0061690 | 1.272 | 0.047 |
| LysoPE (P-18:1(9Z)/0:0) | HMDB0240599 | 1.276 | 0.038 |
| Riboflavin (Vitamin B2) | HMDB0000244 | 1.293 | 0.031 |
| DL-Citrulline | HMDB0000904 | 1.336 | 0.002 |
| Cytidine 2'-phosphate | HMDB0011692 | 1.349 | 0.032 |
| 15beta-Hydroxydesogestrel | HMDB0060707 | 1.380 | 0.045 |
| Prostaglandin F1a | HMDB0002685 | 1.409 | 0.018 |
| PE (20:4(5Z,8Z,11Z,14Z)/0:0) | HMDB0011517 | 1.491 | 0.015 |
| PA (18:0/16:1(9Z)) | HMDB0114877 | 1.497 | 0.003 |
| Gamithromycin | HMDB0252606 | 1.632 | 0.001 |
| Indoxyl Sulfate | HMDB0000682 | 1.910 | 0.047 |
| SM (d16:1/PGJ2) | HMDB0290269 | 2.071 | 0.002 |
| 2-Phenylethanol glucuronide | HMDB0010350 | 9.858 | 0.046 |
Table 2 Disturbed metabolites with P < 0.05 in the high-dose SJGB decoction-treated group compared to the model group
| Metabolite | HMDB ID/ PubChem CIDa | Fold change SJGB vs M | P value SJGB vs M |
|---|---|---|---|
| PC (18:0/18:2(9Z,12Z)) | HMDB0008039 | 0.664 | 0.001 |
| PC (18:2(9Z,12Z)/20:3(5Z,8Z,11Z)) | HMDB0008139 | 0.669 | 0.007 |
| PC (38:4) | HMDB0007988 | 0.671 | 0.024 |
| PS (15:0/24:1(15Z)) | HMDB0112341 | 0.677 | 0.015 |
| PC (18:0/20:4(8Z,11Z,14Z,17Z)) | HMDB0008049 | 0.693 | 0.026 |
| PE-NMe (20:4(8Z,11Z,14Z,17Z)/20:0) | HMDB0113462 | 0.708 | 0.037 |
| PC (36:4) | HMDB0007889 | 0.763 | 0.041 |
| PC (34:1) | HMDB0007879 | 0.803 | 0.047 |
| Thioetheramide-pc | PubChem CID: 10259406 | 0.828 | 0.020 |
| Galactosylhydroxylysine | HMDB0000600 | 0.843 | 0.046 |
| 2-Hydroxyadipic acid | HMDB0000321 | 0.862 | 0.000 |
| Floionolic acid | HMDB0034295 | 1.064 | 0.034 |
| 4-Nitrophenol | HMDB0001232 | 1.077 | 0.039 |
| 5b-Cholestane-3a,7a,12a,23R,25-pentol | HMDB0000513 | 1.153 | 0.021 |
| Glycocholic Acid | HMDB0000331 | 1.195 | 0.010 |
| 2-Hydroxy-6-Pentadecylbenzoic Acid | HMDB0029683 | 1.211 | 0.023 |
| Astromicin | HMDB0248678 | 1.223 | 0.049 |
| N-Glycolylneuraminic acid | HMDB0000833 | 1.228 | 0.040 |
| 5'-Adenylic acid | HMDB0000045 | 1.252 | 0.020 |
| Riboflavin | HMDB0000244 | 1.267 | 0.025 |
| LysoPI (20:4(5Z,8Z,11Z,14Z)/0:0) | HMDB0061690 | 1.272 | 0.047 |
| LysoPE (P-18:1(9Z)/0:0) | HMDB0240599 | 1.276 | 0.038 |
| Riboflavin (Vitamin B2) | HMDB0000244 | 1.293 | 0.031 |
| DL-Citrulline | HMDB0000904 | 1.336 | 0.002 |
| Cytidine 2'-phosphate | HMDB0011692 | 1.349 | 0.032 |
| 15beta-Hydroxydesogestrel | HMDB0060707 | 1.380 | 0.045 |
| Prostaglandin F1a | HMDB0002685 | 1.409 | 0.018 |
| PE (20:4(5Z,8Z,11Z,14Z)/0:0) | HMDB0011517 | 1.491 | 0.015 |
| PA (18:0/16:1(9Z)) | HMDB0114877 | 1.497 | 0.003 |
| Gamithromycin | HMDB0252606 | 1.632 | 0.001 |
| Indoxyl Sulfate | HMDB0000682 | 1.910 | 0.047 |
| SM (d16:1/PGJ2) | HMDB0290269 | 2.071 | 0.002 |
| 2-Phenylethanol glucuronide | HMDB0010350 | 9.858 | 0.046 |
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