Fufang Muji Granules Ameliorate Liver Fibrosis by Reducing Oxidative Stress and Inflammation, Inhibiting Apoptosis, and Modulating Overall Metabolism
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemical Compounds and Reagents
2.2. Determination of Chemical Composition and Content
2.3. Animal Study
2.4. Histopathological Analysis
2.5. Serum and Liver Biochemistry Analysis
2.6. Immunohistochemistry Analysis
2.7. TUNEL Analysis
2.8. HPLC-MS/MS-Based Metabolomics
2.8.1. Sample Preparation
2.8.2. HPLC-MS/MS Method
2.8.3. Data Analysis
2.9. Quantitative Real-Time PCR (qPCR) Analysis
2.10. Determination of Bile Acid Levels in Serum
2.11. Statistical Analysis
3. Results
3.1. Ingredient Identification and Quality Control Results of FMGs
3.2. FMGs Alleviate Hepatic Pathological and Functional Changes in Liver Fibrosis
3.3. FMGs Ameliorate Liver Fibrosis in Rats
3.4. FMGs Ameliorate Myofibroblast Activation in Liver Fibrotic Rats
3.5. FMGs Ameliorate Oxidative Stress Caused by Liver Fibrosis in Rats
3.6. FMGs Reduce Inflammatory Factors in Hepatic Fibrotic Rats
3.7. FMGs Reverse the Metabolic Profiles of Hepatic Fibrotic Rats
3.8. Metabolic Pathway Analysis
3.9. FMGs Regulate Taurine and Hypotaurine Metabolism in Liver Fibrotic Rats
3.10. FMGs Regulate Bile Metabolism in Liver Fibrotic Rats
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene | Forward Primer (5′-3′) | Reverse Primer (5′-3′) |
---|---|---|
TNF-α | AAGGACACCATGAGCACTGAAAGC | AGGAAGGAGAAGAGGCTGAGGAAC |
TGF-β1 | TATTGAGCACCTTGGGCACTGTTG | CCTTAACCTCTCTGGGCTTGTTTCC |
IL-1β | GGACAGGATATGGAGCAACAAGTGG | TCATCTTTCAACACGCAGGACAGG |
IL-6 | GAAACCGCTATGAAGTTCCTCTCTG | GTATCCTCTGTGAAGTCTCCTCTCC |
α-SMA | CAGGGAGTAATGGTTGGAATGGG | AGTTGGTGATGATGCCGTGTTC |
Smad4 | CTGTTGTGACTGTGGATGGCTATG | CTCTCAATCGCTTCTGTCCTGTG |
Smad7 | AAGAGGCTGTGTTGCTGTGAATC | ATTGGGTATCTGGAGTAAGGAGGAG |
FXR | ACCTCGGCTCCTTCTCAGTTG | GGTGGCTGTGGTGAAGACTAATC |
CYP7A1 | ACAGAGGCCCATAGCATCCC | ACAAGGCAAAGCAGGAAGCA |
CSAD | TCATCACGGAGAGCCTCAACAC | GGAGCCACCAGGACAGAAGAC |
Mrp2 | TAGCCTCATTCAGACGACCATCC | TTCAGGACTGCCGTATTCAACAATC |
Bsep | GAAGCCATTGCCGACCAGATG | TGAGAGGACTGACAGCGAGAATC |
CDO | GATTCCATTGGCTTACACCGAGTAG | TTTATGCCCTGTTCTCTGGTCAAAG |
Bile Acids | Abbreviation | Parent Ion (m/z) | Daughter Ion (m/z) | DP (Volts) | CE (Volts) |
---|---|---|---|---|---|
α-Muricholic Acid | α-MAC | 407.4 | 371.2 | −50 | −46 |
Cholic acid | CA | 407.4 | 343.1 | −60 | −47 |
Chenodeoxycholic acid | CDCA | 391.3 | 391.3 | −160 | −60 |
Deoxycholic acid | DCA | 391.4 | 345.1 | −60 | −44 |
Glycocholic acid | GCA | 464.4 | 464.4 | −50 | −8 |
Glydeoxycholic acid | GDCA | 448.4 | 448.4 | −150 | −8 |
Taurocholic acid | TCA | 514.4 | 79.9 | −270 | −120 |
Taurochenodeoxycholic acid | TCDCA | 498.4 | 123.9 | −43 | −66 |
Tauroursodeoxycholic acid | TUDCA | 498.4 | 79.8 | −80 | −120 |
Tauro-α-muricholic acid | T-α-MAC | 514.4 | 514.4 | −41 | −15 |
Bile Acids | Retention Time (min) | Calibration Curves | Correlation Coefficient (r2) | Linear Range (ng/mL) |
---|---|---|---|---|
α-MAC | 3.25 | y = 6.090 × 10−3x + 6.530 × 10−3 | 0.9925 | 1–1000 |
CA | 4.09 | y = 2.123 × 10−2x + 4.413 × 10−2 | 0.9946 | 1–1000 |
CDCA | 5.57 | y = 0.2421x + 2.572 | 0.9945 | 1–1000 |
DCA | 5.78 | y = 3.328 × 10−2x + 3.209 × 10−2 | 0.9926 | 1–1000 |
GCA | 3.13 | y = 0.1203x + 0.1686 | 0.9951 | 1–1000 |
GDCA | 4.22 | y = 0.1828x + 0.3437 | 0.9907 | 1–1000 |
TCA | 2.57 | y = 2.894 × 10−2x + 3.643 × 10−2 | 0.9910 | 1–1000 |
TCDCA | 3.36 | y = 1.461 × 10−2x + 1.598 × 10−2 | 0.9905 | 1–1000 |
TUDCA | 2.51 | y = 9.325 × 10−2x + 1.062 × 10−2 | 0.9922 | 1–1000 |
T-α-MAC | 1.95 | y = 0.1631x + 0.1663 | 0.9908 | 1–1000 |
Bile Acids | Control (ng/mL) | Model (ng/mL) | FMG (ng/mL) |
---|---|---|---|
α-MAC | 836.4 ± 398.7 | 3686 ± 306.3 ### | 33.19 ± 22.42 *** |
CA | 3700 ± 1877 | 22,973 ± 2894 ### | 368.1 ± 138.3 *** |
CDCA | 1110 ± 636.3 | 2365 ± 34.49 | 33.20 ± 14.90 |
DCA | 389.6 ± 187.5 | 229.9 ± 57.5 | 54.02 ± 20.2 |
GCA | 1411 ± 659.6 | 6669 ± 1532 ## | 983.3 ± 446.4 ** |
GDCA | 250.2 ± 125.1 | 908.3 ± 350.1 ## | 223.5 ± 24.2 * |
TCA | 3155 ± 841.2 | 26,283 ± 1365 ### | 1397 ± 561.5 *** |
TCDCA | 330.2 ± 159.8 | 1499 ± 272.6 ### | 131.4 ± 159.8 ** |
TUDCA | 174.8 ± 79.0 | 200.9 ± 57.1 | 131.4 ± 18.0 |
T-α-MAC | 806.4 ± 386.4 | 3696 ± 1216 ## | 643.0 ± 198.9 * |
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Men, L.; Gu, Z.; Wang, E.; Li, J.; Li, Z.; Li, K.; Li, C.; Gong, X. Fufang Muji Granules Ameliorate Liver Fibrosis by Reducing Oxidative Stress and Inflammation, Inhibiting Apoptosis, and Modulating Overall Metabolism. Metabolites 2024, 14, 446. https://doi.org/10.3390/metabo14080446
Men L, Gu Z, Wang E, Li J, Li Z, Li K, Li C, Gong X. Fufang Muji Granules Ameliorate Liver Fibrosis by Reducing Oxidative Stress and Inflammation, Inhibiting Apoptosis, and Modulating Overall Metabolism. Metabolites. 2024; 14(8):446. https://doi.org/10.3390/metabo14080446
Chicago/Turabian StyleMen, Lei, Zhihong Gu, Enhua Wang, Jiwen Li, Zhongyu Li, Keke Li, Chunbin Li, and Xiaojie Gong. 2024. "Fufang Muji Granules Ameliorate Liver Fibrosis by Reducing Oxidative Stress and Inflammation, Inhibiting Apoptosis, and Modulating Overall Metabolism" Metabolites 14, no. 8: 446. https://doi.org/10.3390/metabo14080446