The Root of Polygonum multiflorum Thunb. Alleviates Non-Alcoholic Steatosis and Insulin Resistance in High Fat Diet-Fed Mice
<p>Cytotoxicity of PM in HepG2 cells. (<b>A</b>) HepG2 cells were treated with PM at the indicated concentrations for 24 h, and cell viability was measured by MTT assay. Data are the mean ± SEM from three independent experiments. ** <span class="html-italic">p</span> < 0.01 vs. CTL (control group). (<b>B</b>) The cells were seeded in E-plate 16, and proliferation, migration, and adherence were monitored every 15 min in the real time cell analyzer (RTCA) for 48 h. When the cells reached the logarithmic growth phase, PM was treated at the concentrations of 0, 0.05, 0.1, or 0.2 mg/mL in quadruplicate, and cell viability was monitored by continuous impedance recording every 15 min in RTCA.</p> "> Figure 2
<p>The effect of PM on intracellular lipid and triglyceride (TG) accumulation in free fatty acid (FFA)-treated HepG2 cells. Representative pictures (magnification 200X) of Oil-red O stain in HepG2 cells from three independent experiments (<b>A</b>), the quantification of lipid contents (<b>B</b>), and intracellular TG levels (<b>C</b>) were shown. Control groups were treated with 1% of fatty acid-free bovine serum albumin (BSA), and FFA groups were treated with 1 mM of FFA for 24 h, and then were harvested. PM was pre-treated 1 h prior to FFA at the indicated concentrations. Data are the mean ± SEM. ** <span class="html-italic">p</span> < 0.01 vs. Veh + control group; <sup>##</sup> <span class="html-italic">p</span> < 0.01 vs. Veh + FFA group.</p> "> Figure 3
<p>The effect of PM extract on the adenosine monophosphate-activated kinase (AMPK) and acetyl-CoA carboxylase (ACC) phosphorylation, glucose transporter 4 (GLUT4), and sterol regulatory element-binding protein-1 (SREBP-1) protein levels in FFA-treated HepG2 cells. Representative images of western blot analysis were from three independent experiments, and the densitometric quantification of relative band intensities are presented. The cells were treated with 1 mM of FFA for 24 h, and then were harvested. PM was pretreated 1 h prior to FFA treatment at the indicated concentrations, and control cells were treated with 1% of fatty acid-free BSA. Data are the mean ± SEM. * <span class="html-italic">p</span> < 0.05 and ** <span class="html-italic">p</span> < 0.01 vs. Veh + control group; <sup>#</sup> <span class="html-italic">p</span> < 0.05 and <sup>##</sup> <span class="html-italic">p</span> < 0.01 vs. Veh + FFA group.</p> "> Figure 4
<p>The effect of PM extract on body weight, food intake, and weight of adipose tissues in normal diet (ND)- or high-fat diet (HFD)-fed mice. The mice were administered PM extract or vehicle orally for 16 weeks, and the adipose tissues were collected. Body weight (<b>A</b>) and food intake (<b>B</b>) were measured every 10 days for 16 weeks. At sacrifice, representative images of epididymal and perirenal fats were taken, and the weight of these fats were measured. Data are the mean ± SEM. * <span class="html-italic">p</span> < 0.05 and ** <span class="html-italic">p</span> < 0.01 vs. Veh + ND group; <sup>#</sup> <span class="html-italic">p</span> < 0.05 and <sup>##</sup> <span class="html-italic">p</span> < 0.01 vs. Veh + HFD group.</p> "> Figure 5
<p>The effect of PM extract on hepatic triglyceride (TG) and lipid accumulation in HFD-fed mice. Hepatic TG levels (<b>A</b>) and representative images (magnification 200X) of H&E (<b>B</b>) and Oil-red O staining (<b>C</b>). The mice fed with ND or HFD were treated with PM extract or vehicle for 16 weeks, and the liver tissues were collected. Data are the mean ± SEM. * <span class="html-italic">p</span> < 0.05 vs. Veh + ND group; <sup>#</sup> <span class="html-italic">p</span> < 0.05 vs. Veh + HFD group.</p> "> Figure 6
<p>The effect of PM extract on fasting blood glucose levels, glucose tolerance, and insulin sensitivity in HFD-fed mice. The mice fed with ND or HFD were treated with PM extract or vehicle. The mice fasted for 16 h, and their blood glucose levels were measured every 2 weeks. For the glucose tolerance test (GTT), the mice fasted for 16 h before assaying. Fasting blood glucose levels (<b>A</b>), GTT performed after injecting 20% D-glucose (2 g/kg body weight) (<b>B</b>), and insulin tolerance test (ITT) performed after injecting insulin (1 U/kg body weight) (<b>C</b>). Data are the mean ± SEM. * <span class="html-italic">p</span> < 0.05 vs. Veh + ND group; <sup>#</sup> <span class="html-italic">p</span> < 0.05 vs. Veh + HFD group.</p> "> Figure 7
<p>The effect of PM extract on AMPK and ACC phosphorylation, GLUT4, and SREBP-1 protein levels in the liver tissues of ND- or HFD-fed mice. Representative images of western blot analysis are shown, and densitometric quantifications of relative band intensities are presented. The mice fed with ND or HFD were treated with PM extract or vehicle for 16 weeks, and the liver tissues were collected. Data are the mean ± SEM. * <span class="html-italic">p</span> < 0.05 and ** <span class="html-italic">p</span> < 0.01 vs. Veh + ND group; <sup>#</sup> <span class="html-italic">p</span> < 0.05 and <sup>##</sup> <span class="html-italic">p</span> < 0.01 vs. Veh + HFD group.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Preparation of PM Extract
2.2. High-Performance Liquid Chromatography (HPLC)
2.3. Animals and Treatments
2.4. Cell Culture and Treatments
2.5. Cytotoxicity of PM
2.6. Measurement of Blood Glucose Levels
2.7. Glucose Tolerance Test and Insulin Tolerance Test
2.8. Triglyceride Levels in Liver Tissues and HepG2 Cells
2.9. H&E and Oil Red O Staining of Liver Tissues
2.10. Western Blot Analysis
2.11. Statistical Analysis
3. Results
3.1. Phytochemical Contents in the PM Extract
3.2. Cytotoxicity of PM in HepG2 Cells
3.3. PM Extract Attenuated the Increases of Lipid Accumulation and Intracellular TG Levels in FFA-Exposed HepG2 Cells
3.4. PM Extract Modulated Lipogenic and Lipolytic Protein Levels in FFA-Exposed HepG2 Cells
3.5. PM Extract Attenuated the Weight Increase of Adipose Tissues without Changing Body Weight or Food Intake in HFD-Fed Mice
3.6. PM Extract Attenuated the Increases of Hepatic TG Levels and Hepatocellular Lipid Accumulation in HFD-Fed Mice
3.7. PM Extract Reduced Fasting Blood Glucose Levels, Improved Glucose Tolerance, and Insulin Sensitivity in HFD-Fed Mice
3.8. PM Extract Modulated Lipogenic and Lipolytic Protein Levels in HFD-Fed Mice
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Phytochemical | Content 1 (mg/g) |
---|---|
Catechin | 1.51 ± 0.07 |
2,3,5,4′-tetrahydroxystilbene-2-O-α-glucoside (TSG) | 36.68 ± 1.83 |
Rhein | 0.3 ± 0.02 |
Emodin | 0.05 ± 0.00 |
Chrysophenol | ND 2 |
Total | 38.54 ± 1.93 |
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Jung, S.; Son, H.; Hwang, C.E.; Cho, K.M.; Park, S.W.; Kim, H.; Kim, H.J. The Root of Polygonum multiflorum Thunb. Alleviates Non-Alcoholic Steatosis and Insulin Resistance in High Fat Diet-Fed Mice. Nutrients 2020, 12, 2353. https://doi.org/10.3390/nu12082353
Jung S, Son H, Hwang CE, Cho KM, Park SW, Kim H, Kim HJ. The Root of Polygonum multiflorum Thunb. Alleviates Non-Alcoholic Steatosis and Insulin Resistance in High Fat Diet-Fed Mice. Nutrients. 2020; 12(8):2353. https://doi.org/10.3390/nu12082353
Chicago/Turabian StyleJung, Soonwoong, Hyeonwi Son, Chung Eun Hwang, Kye Man Cho, Sang Won Park, Hwajin Kim, and Hyun Joon Kim. 2020. "The Root of Polygonum multiflorum Thunb. Alleviates Non-Alcoholic Steatosis and Insulin Resistance in High Fat Diet-Fed Mice" Nutrients 12, no. 8: 2353. https://doi.org/10.3390/nu12082353