Heat Killed Lactobacillus reuteri GMNL-263 Reduces Fibrosis Effects on the Liver and Heart in High Fat Diet-Hamsters via TGF-β Suppression
"> Figure 1
<p>The animal epididymal adipose tissue weight and liver lipid and cholesterol metabolism biomarkers. (<b>A</b>) The HFD hamster epididymal adipose tissue weights were higher than in the control group, and the apididymal adipose tissue weight was lower in the heat-killed Lr263 (625 and 3125 mg/kg-hamster/day) treatment groups; (<b>B</b>) FAS; (<b>C</b>) HMG-CoA reductase; (<b>D</b>) CYP7A1; (<b>E</b>) PPARγ; and (<b>F</b>) LDLR are liver lipid metabolism biomarkers and were analyzed using RT-PCR. FAS and HMG-CoA reductase were greater in the HFD group and lower in the heat killed Lr263 (3125 mg/kg-hamster/day) treatment groups, but the difference in HMG-CoA reductase was not significant in the heat-killed Lr263 treatment groups. LDLR and CYP7A1 was lower in the HFD group and greater in the heat killed Lr263 (625 and 3125 mg/kg-hamster/day) treatment groups. (* <span class="html-italic">p</span> < 0.05 compared with the HFD group).</p> "> Figure 2
<p>Fibrosis assessments. Cirrhosis was assessed using Massion’s trichrome (MS) staining assay to indicate the collagen accumulation (by the blue color) in liver tissue slides. The bar length is 100 μm.</p> "> Figure 3
<p>TGF-β protein levels in animal livers. (<b>A</b>) High TGF-β expression was observed only in the HFD group hamster livers; (<b>B</b>) The normalized protein expression of TGF-β with β-actin (* <span class="html-italic">p</span> < 0.05 compared with the control group, # <span class="html-italic">p</span> < 0.05, and ### <span class="html-italic">p</span> < 0.001 compared with the HFD group).</p> "> Figure 4
<p>The echocardiography analysis of the hamster was performed using a 10 MHz transducer (GE 10S-RS). The B-mode was visualized for two-dimensional (long-axis and short-axis of the left ventricle) mode images and B-mode perspectives were further used to evaluate the left ventricle for the M-mode cursor. An M-mode evaluation of heart function was performed by comparing the left ventricular systolic and diastolic distances, which are shown as ejection fraction (EF) values and fractional shortening (FS) values (<span class="html-italic">n</span> = 6 in each group, * <span class="html-italic">p</span> < 0.05 compared with control group, and # <span class="html-italic">p</span> < 0.05 compared with HFD group).</p> "> Figure 5
<p>Assessment of fibrosis in the animal hearts. Cirrhosis was assessed using Masson’s trichrome (MS) staining assay to indicate collagen accumulation (the blue color indicated by the arrows) in the heart tissue slides. The bar length is 100 μm.</p> "> Figure 6
<p>TGF-β pathway protein expression analysis. (<b>A</b>) The TGF-β/Smad-3/CTGF expression levels increased in the HFD treatment hamster hearts and decreased in the heat-killed Lr263 (625 and 3125 mg/kg-hamster/day) treatment hamster hearts; (<b>B</b>) The normalized protein expression of TGF-β/β-actin, p-Erk/Erk, p-Smad3/Smad3, and CTGF/β-actin (*** <span class="html-italic">p</span> < 0.001 compared with the control group, # <span class="html-italic">p</span> < 0.05, and ### <span class="html-italic">p</span> < 0.001 compared with the HFD group).</p> ">
Abstract
:1. Introduction
2. Results and Discussion
Treatments | Control | HFD | HFD + Lr-263(L) | HFD + Lr-263(H) |
---|---|---|---|---|
Parameter | ||||
Body Weight (g) | 126.5 ± 10.6 * | 138.3 ± 5.9 | 136.7 ± 3.8 | 124.7 ± 4.0 * |
Food Intake (g) | 8.1 ± 0.8 | 8.2 ± 0.9 | 7.9 ± 1.1 | 7.8 ± 0.5 |
Liver | ||||
TG (mg/g) | 85.0 ± 6.5 * | 93.3 ± 5.3 | 90.7 ± 3.4 | 84.0 ± 4.3 * |
T-CHO (mg/g) | 89.7 ± 0.4 * | 130.0 ± 4.9 | 118.3 ± 12.8 | 127.3 ± 6.0 |
MDA (μg/g) | 4.2 ± 0.4 * | 8.6 ± 0.7 | 6.7 ± 1.1 | 5.5 ± 0.4 * |
Fecal | ||||
TG (mg/g) | 5.9 ± 0.7 | 4.7 ± 0.8 | 7.6 ± 0.9 * | 10.4 ± 1.7 * |
T-CHO (mg/g) | 6.7 ± 0.7 | 8.5 ± 1.4 * | 8.5 ± 1.4 | 11.0 ± 0.8 * |
3. Experimental Section
3.1. Preparing the Probiotic Suspensions
3.2. Animals
Components (g/kg) | Normal Diet | High-Fat Diet |
---|---|---|
Casein | 200 | 232 |
l-Cystine | 3.0 | 3.0 |
d,l-Methionine | N.D. | 3.5 |
Corn Starch | 397.48 | 137 |
Maltodextrin | 132 | 150 |
Sucrose | 100 | 162.58 |
Cellulose | 50 | 50 |
Cholesterol | N.D. | 1.9 |
Mineral Mix (AIN-93) | 35 | 40.60 |
Calcium phosphate dibasic | N.D. | 4.64 |
Vitamin Mix (AIN-93) | 10 | 16.24 |
Choline Bitartrate | 2.5 | 5 |
Tert-butylhydroquinone | 0.014 | 0.04 |
Soybean oil | 70 | 40 |
Lard | N.D. | 153.5 |
3.3. Cardiac Echocardiography
3.4. Masson’s Trichrome Staining
3.5. RNA Extraction and RT-PCR
3.6. Tissue Protein Extraction
3.7. Western Blot Assay
3.8. Liver and Fecal Lipid Profile Analysis
3.9. Statistical Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Ting, W.-J.; Kuo, W.-W.; Hsieh, D.J.-Y.; Yeh, Y.-L.; Day, C.-H.; Chen, Y.-H.; Chen, R.-J.; Padma, V.V.; Chen, Y.-H.; Huang, C.-Y. Heat Killed Lactobacillus reuteri GMNL-263 Reduces Fibrosis Effects on the Liver and Heart in High Fat Diet-Hamsters via TGF-β Suppression. Int. J. Mol. Sci. 2015, 16, 25881-25896. https://doi.org/10.3390/ijms161025881
Ting W-J, Kuo W-W, Hsieh DJ-Y, Yeh Y-L, Day C-H, Chen Y-H, Chen R-J, Padma VV, Chen Y-H, Huang C-Y. Heat Killed Lactobacillus reuteri GMNL-263 Reduces Fibrosis Effects on the Liver and Heart in High Fat Diet-Hamsters via TGF-β Suppression. International Journal of Molecular Sciences. 2015; 16(10):25881-25896. https://doi.org/10.3390/ijms161025881
Chicago/Turabian StyleTing, Wei-Jen, Wei-Wen Kuo, Dennis Jine-Yuan Hsieh, Yu-Lan Yeh, Cecilia-Hsuan Day, Ya-Hui Chen, Ray-Jade Chen, Viswanadha Vijaya Padma, Yi-Hsing Chen, and Chih-Yang Huang. 2015. "Heat Killed Lactobacillus reuteri GMNL-263 Reduces Fibrosis Effects on the Liver and Heart in High Fat Diet-Hamsters via TGF-β Suppression" International Journal of Molecular Sciences 16, no. 10: 25881-25896. https://doi.org/10.3390/ijms161025881