Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model
Abstract
:1. Introduction
2. Results
2.1. Effects of Arazyme on Metabolic Parameters
2.2. Effects of Arazyme on Plasma Biomarkers
2.3. Arazyme Reduced HFD-Induced Hepatic Steatosis
2.4. Arazyme Inhibited Lipid Accumulation in HepG2 Cells
2.5. Arazyme Suppressed Steatohepatitis and Macrophage Recruitment
2.6. Arazyme Inhibited PA-Induced Inflammation in RAW264.7 Macrophages
3. Discussion
4. Materials and Methods
4.1. Animals and Diets
4.2. Measurement of Metabolic Parameters
4.3. Histological Analysis of the Liver
4.4. Real-Time Quantitative RT-PCR (qRT-PCR)
4.5. Western Blotting Analysis
4.6. Experiments in HepG2 Cells and RAW264.7 Cells
4.7. Statistical Analyses
Author Contributions
Funding
Conflicts of Interest
References
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ND | HFD | HFD+Ara | HFD+MT | |
---|---|---|---|---|
Initial body weight (g) | 23.5 ± 0.3 | 23.6 ± 0.3 | 23.3 ± 0.4 | 23.5 ± 0.4 |
Final body weight (g) | 38.8 ± 0.8 ‡ | 44.9 ± 1.4 ‡, ## | 44.4 ± 0.8 ‡ | 43.6 ± 1.9 ‡ |
Weight gain (g) | 15.5 ± 0.5 | 21.3 ± 1.2 ## | 21.5 ± 0.7 | 20.2 ± 1.5 |
Food intake (g/day) | 2.3 ± 0.1 | 2.0 ± 0.2 # | 2.0 ± 0.1 | 2.1 ± 0.1 |
Liver (g) | 1.28 ± 0.04 | 1.36 ± 0.09 | 1.29 ± 0.06 | 1.35 ± 0.13 |
ND | HFD | HFD+Ara | HFD+MT | |
---|---|---|---|---|
Glucose | ||||
Initial (mg/dL) | 99.9 ± 3.5 | 100.6 ± 3.7 | 99.5 ± 2.6 | 100.9 ± 4.3 |
Final (mg/dL) | 126.4 ± 10.3 † | 169.0 ± 12.9 ‡, # | 145.8 ± 7.9 ‡ | 152.5 ± 14.7 ‡ |
Insulin | ||||
Initial (ng/mL) | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 |
Final (ng/mL) | 0.9 ± 0.2 ‡ | 2.3 ± 0.5 ‡, # | 2.5 ± 0.4 ‡ | 2.3 ± 0.4 ‡ |
HbA1c (%) | 5.05 ± 0.09 | 5.41 ± 0.12 # | 5.15 ± 0.06 | 5.12 ± 0.13 * |
HOMA-IR | 6.9 ± 1.3 | 19.1 ± 4.7 # | 16.4 ± 3.0 | 17.5 ± 4.3 |
TG (mg/dL) | 118.2 ± 7.5 | 142.7 ± 9.5 # | 118.4 ± 4.2 * | 111.7 ± 10.0 ** |
TC (mg/dL) | 144.5 ± 6.4 | 183.1 ± 9.4 # | 170.6 ± 7.2 | 162.0 ± 13.0 |
HDL-C (mg/dL) | 74.4 ± 6.0 | 73.2 ± 2.6 | 68.0 ± 2.2 | 63.7 ± 3.6 |
HDL-C/TC (%) | 50.7 ± 3.5 | 40.9 ± 2.0 ## | 39.7 ± 1.5 | 40.4 ± 1.6 |
NEFA (mEq/L) | 2.1 ± 0.1 | 2.7 ± 0.1 # | 2.3 ± 0.1 * | 2.2 ± 0.2 * |
AST (IU/L) | 96.5 ± 6.1 | 117.4 ± 5.8 # | 105.8 ± 11.3 | 111.8 ± 10.4 |
ALT (IU/L) | 26.0 ± 3.6 | 45.3 ± 10.2 # | 41.5 ± 5.7 | 40.2 ± 4.2 |
Gene (Number) | Forward Primer | Reverse Primer |
---|---|---|
Acc1 (NM_133360) | AGTTTCCCAGCCAGCAGATT | ATCCATCACCACAGCCTTCA |
Acc2 (NM_133904) | CCCATCACCACTCCTTCTGA | GTCCGAGTCTCCACAGCAAT |
Acox (NM_015729) | GCTGGGCTGAAGGCTTTTACTA | AATCCCACTGCTGTGAGAATAGC |
Ccl2 (NM_011333) | TCACCTGCTGCTACTCATTC | TACAGAAGTGCTTGAGGTGG |
Ccl3 (NM_011337) | CACCCTCTGTCACCTGCTCAA | TGGCGCTGAGAAGACTTGGT |
Ccl4 (NM_013652) | CTAACCCCGAGCAACACCAT | AGCCCATTGGTGCTGAGAAC |
Ccl5 (NM_013653) | TCCCTGTCATTGCTTGCTCTAG | GAGCAGCTGAGATGCCCATT |
Cpt1α (NM_013495) | CTGCACTCCTGGAAGAAGAA | GTTCTTCGTCTGGCTTGACA |
Dgat1 (NM_010046) | ACAACCTGACCTACCGAGAT | AGTAGGGACCATCCACTGTT |
Dgat2 (NM_026384) | GCTGGCATTTGACTGGAACA | TGGTCAGCAGGTTGTGTGTCTT |
Fas (NM_007988) | TGTGAGTGGTTCAGAGGCAT | TTCTGTAGTGCCAGCAAGCT |
Gapdh (NM_001001303) | ACATCATCCCTGCATCCACT | AGATCCACGACGGACACATT |
Gpam (NM_001356285) | CCGGAAGAGGCCCTTCGTGG | GCCAGCCATCCTCTGTGCCT |
Il1b (NM_008361) | ATGGCAACTGTTCCTGAACTCAACT | ATATTCTGTCCATTGAGGTGGAGAGCT |
Mlxipl (NM_021455) | CAGATGCGGGACATGTTTGA | AATAAAGGTCGGATGAGGATGCT |
Pparα (NM_011144) | CCTGAACATCGAGTGTCGAA | GTACTGGCATTTGTTCCGGT |
Scd1 (NM_009127) | ACGCCGACCCTCACAATTC | AGTTTTCCGCCCTTCTCTTTG |
Scd2 (NM_009128) | CCGTGGCTTCTTTTTCTCTCA | TTCCGCCCTTCTCTTTGACA |
Srebf1 (NM_011480) | GAGCGAGCGTTGAACTGTAT | ATGCTGGAGCTGACAGAGAA |
Srebf2 (NM_033218) | TCCTCCATCAACGACAAAATCA | ACTTGTGCATCTTGGCATCTGT |
Tnfa (NM_001278601) | CTCAGATCATCTTCTCAAAATTCGAGTGACA | CTTCACAGAGCAATGACTCCAAAGT |
Ucp2 (NM_011671) | GCCTCTACGACTCTGTCAAA | CTTCGACAGTGCTCTGGTAT |
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Li, H.; Yoo, W.; Park, H.-M.; Lim, S.-Y.; Shin, D.-H.; Kim, S.; Park, H.-Y.; Jeong, T.-S. Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model. Int. J. Mol. Sci. 2019, 20, 2325. https://doi.org/10.3390/ijms20092325
Li H, Yoo W, Park H-M, Lim S-Y, Shin D-H, Kim S, Park H-Y, Jeong T-S. Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model. International Journal of Molecular Sciences. 2019; 20(9):2325. https://doi.org/10.3390/ijms20092325
Chicago/Turabian StyleLi, Hua, Wonbeak Yoo, Hye-Mi Park, Soo-Youn Lim, Dong-Ha Shin, Seokho Kim, Ho-Yong Park, and Tae-Sook Jeong. 2019. "Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model" International Journal of Molecular Sciences 20, no. 9: 2325. https://doi.org/10.3390/ijms20092325