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. 2024 Jan 19;16(2):305.
doi: 10.3390/nu16020305.

Effects of Fermented Polygonum cuspidatum on the Skeletal Muscle Functions

Young-Seon Kim  1   2   3 , Ji-Hye Han  3 , Chang-Hoon Lim  1   2   4 , Xue-Quan Fang  1   2   4 , Hyeock-Soon Jang  3 , Sang-Yun Lee  3 , Woo-Jong Yim  3 , Ji-Hong Lim  1   2   4
Affiliations

Effects of Fermented Polygonum cuspidatum on the Skeletal Muscle Functions

Young-Seon Kim et al. Nutrients. .

Abstract

Plant extract fermentation is widely employed to enhance the nutritional and pharmaceutical value of functional foods. Polygonum cuspidatum (Pc) contains flavonoids, anthraquinones, and stilbenes, imparting protective effects against inflammatory diseases, cancer, diabetes, and cardiovascular diseases. However, the effects of fermented Pc on skeletal muscle strength remain unexplored. In this study, we generated fermented Pc using a complex of microorganisms containing Lactobacillus spp. (McPc) and assessed its effects on muscle strength and motor function in mice. Compared to unfermented Pc water extract, elevated levels of emodin and resveratrol were noted in McPc. This was identified and quantified using UPLC-QTOF/MS and HPLC techniques. Gene expression profiling through RNA-seq and quantitative RT-PCR revealed that McPc administration upregulated the expression of genes associated with antioxidants, glycolysis, oxidative phosphorylation, fatty acid oxidation, and mitochondrial biogenesis in cultured C2C12 myotubes and the gastrocnemius muscle in mice. McPc significantly improved skeletal muscle strength, motor coordination, and traction force in mice subjected to sciatic neurectomy and high-fat diet (HFD). McPc administration exhibited more pronounced improvement of obesity, hyperglycemia, fatty liver, and hyperlipidemia in HFD mice compared to control group. These findings support the notion that emodin and resveratrol-enriched McPc may offer health benefits for addressing skeletal muscle weakness.

Keywords: Polygonum cuspidatum; emodin; fermentation; resveratrol; skeletal muscle.

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Conflict of interest statement

Authors Young-Seon Kim, Ji-Hye Han, Hyeock-Soon Jang, Sang-Yun Lee and Woo-Jong Yim were employed by the company Jung-Ang Microbe Research Institute (JM). The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Qualification and quantification of primary compounds in the Polygonum cuspidatum (Pc) water extract and microbe complex-fermented Polygonum cuspidatum (McPc). (A) Ultra-high performance liquid chromatogram of McPc and Pc. (B) Identification of 10 compounds in Pc and McPc. (C) High-performance liquid chromatogram (HPLC) for quantitative anthraquinones in Pc and McPc. (D) HPLC for quantitative stilbenes in Pc and McPc. (E) Total anthraquinone and stilbene contents of the primary compounds in the stem of Pc.
Figure 2
Figure 2
Transcriptome analysis in McPc-treated C2C12 myotubes. (A) MA plot (log2 fold change vs. average logCPM) of vehicle and McPc. (B) Gene ontology (GO) classification of differentially expressed genes. The graph shows approximately 10 GO terms with most genes annotated. The top 10 GO terms were assigned to the differentially expressed genes in C2C12 myotubes treated with or without McPc. (C) Gene expression of C2C12 myotubes incubated with McPc (μg/mL) for 24 h. (D) The protein level of LPL in C2C12 myotubes treated with various concentrations of McPc for 24 h. (E) Gene expression of C2C12 myotubes incubated with 20 μM emodin or 50 μM resveratrol for 24 h. The values represent the mean ± SD (n = 3); * p < 0.05, ** p < 0.01 and *** p < 0.001. ns = not significant. One-way analysis of variance (ANOVA) Tukey’s post hoc test was performed for statistical analysis.
Figure 3
Figure 3
Antioxidant activity of fermented McPc in vitro and in vivo. (A,B) Antioxidant-related genes, NQO1, GSTA2, and GPX3, mRNA expression of C2C12 myotubes treated with McPc (n = 3) and McPc-diet mice (n = 5). (C) Intracellular reactive oxygen species (ROS) levels of C2C12 myoblasts and myotubes (n = 4). C2C12 cells were treated with 200 μg/mL of McPc in presence or absence of H2O2 (100 μM). (D) Intracellular ROS levels of 100 μg/mL McPc-treated C2C12 myotubes incubated with dexamethasone (100 μM) to induce ROS (n = 4). The values represent the mean ± SD; * p < 0.05, ** p < 0.01, and *** p < 0.001. ns = not significant. One-way ANOVA Tukey’s post hoc test was conducted for multiple comparisons, and the two-tailed unpaired Student’s t-test was conducted for single comparisons.
Figure 4
Figure 4
Expression of genes associated with mitochondrial oxidative capacity in McPc-treated C2C12 myotubes. (A) Expression of mRNA-encoding transcription factors and coactivators related to mitochondrial oxidative capacity in C2C12 myotubes incubated with McPc for 24 h. (B) Glycolysis-related gene expression. (C) Fatty-acid-oxidation-related gene expression. (D) Oxidative phosphorylation (OXPHOS)-associated gene expression. (E) Levels of OXPHOS complex proteins (CI: NDUFB8, CII: SDHB, CIII: UQCRC2, CV: ATP5A) in C2C12 myotubes treated with McPc for 24 h. (F) Protein levels in C2C12 myotubes treated with McPc for 24 h. (G) Mitochondrial content of 100 μg/mL McPc-treated C2C12 myotubes measured using the MitoTracker staining assay. The density of MitoTracker Deep Red FM fluorescent was evaluated using a microplate reader (λex 644 nm and λem 665 nm). The values represent mean ± SD; * p < 0.05, ** p < 0.01, and *** p < 0.001. One-way ANOVA Tukey’s post hoc test was conducted for multiple comparisons and two-tailed unpaired Student’s t-test was conducted for single comparisons.
Figure 5
Figure 5
Skeletal muscle strength and function in McPc-treated mice. (A) Grip strength and (B) rotarod latency in McPc-treated mice. McPc (100 mg/kg) was orally administered once a day for 7 days (n = 5). (C–F) Effect of McPc treatment on (C) grip strength, (D) rotarod latency, (E) gastrocnemius muscle weight, and (F) thickness in sciatic neurectomy (NTX) in mice (n = 5). McPc (75 mg/kg) was orally administered once a day for 28 days from 2 weeks after NTX. (G) The mRNA expression of Mstn, Fbxo32, and Trim63 in vehicle or McPc-treated NTX mice model. (H) Protein levels of muscle differentiation markers, MyoD, myogenin, and MyHC, in C2C12 myotubes incubated with McPc in a dose-dependent manner for 24 h. (I) Brightfield image by differentiation time of C2C12 myotubes treated with 100 μg/mL McPc in differentiation media (2% horse serum). (J) OXPHOS and (K) fatty-acid-oxidation-related gene expression in gastrocnemius muscle of vehicle or McPc-treated NTX mice. (L) Mitochondrial DNA content in gastrocnemius muscle of vehicle or McPc-treated NTX mice. The values represent the mean ± SEM; * p < 0.05, ** p < 0.01, and *** p < 0.001. ns = not significant. One-way ANOVA Tukey’s post hoc test was conducted for multiple comparisons and two-tailed unpaired Student’s t-test was conducted for single comparisons.
Figure 6
Figure 6
McPc rescues impaired skeletal muscle strength and function caused by HFD. (A) Body weight and (B) food intake of male C57BL/6 mice fed a normal chow diet (NCD) or an HFD supplemented with or without McPc over 12 weeks (n = 9). (C) Effect of McPc diet on rotarod latency in NCD and HFD mice. (D) TFAM and ERRγ mRNA expression in the gastrocnemius muscle of non-treated or McPc-treated HFD mice. (E) NQO1, PGC1α, and ACSL1 mRNA expression in the gastrocnemius muscle of non-treated or McPc-treated HFD mice. (F) Tissue weight of liver, inguinal white adipose (ingWAT), and gastrocnemius of NCD and HFD mice supplemented with vehicle or McPc. The values represent the mean ± SEM (n = 9); * p < 0.05, ** p < 0.01, and *** p < 0.001. One-way ANOVA Tukey’s post hoc test for multiple comparisons and two-tailed unpaired Student’s t-test for single comparisons were performed for statistical analysis. (G) Expression of OXPHOS complex proteins in the gastrocnemius muscle of NCD and HFD mice supplemented with vehicle or McPc (n = 6). (H) Densitometric analysis of Western blots revealing OXPHOS (complex I, II, and V) expression. Six independent Western blots were quantified with densitometry using Image J Software Version 1.53. The protein levels were normalized with respect to the corresponding intensities of α-tubulin. The values represent the mean ± SD (n = 6); ** p < 0.01 and *** p < 0.001. ns = not significant. Unpaired Student’s t-test was performed for statistical analysis.
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References

    1. Ke J., Li M.T., Xu S., Ma J., Liu M.Y., Han Y. Advances for pharmacological activities of Polygonum cuspidatum—A review. Pharm. Biol. 2023;61:177–188. doi: 10.1080/13880209.2022.2158349. - DOI - PMC - PubMed
    1. Geng Q., Wei Q., Wang S., Qi H., Zhu Q., Liu X., Shi X., Wen S. Physcion 8-O-beta-glucopyranoside extracted from Polygonum cuspidatum exhibits anti-proliferative and anti-inflammatory effects on MH7A rheumatoid arthritis-derived fibroblast-like synoviocytes through the TGF-beta/MAPK pathway. Int. J. Mol. Med. 2018;42:745–754. doi: 10.3892/ijmm.2018.3649. - DOI - PMC - PubMed
    1. Liu B., Li S., Sui X., Guo L., Liu X., Li H., Gao L., Cai S., Li Y., Wang T., et al. Root Extract of Polygonum cuspidatum Siebold & Zucc. Ameliorates DSS-Induced Ulcerative Colitis by Affecting NF-kappaB Signaling Pathway in a Mouse Model via Synergistic Effects of Polydatin, Resveratrol, and Emodin. Front. Pharmacol. 2018;9:347. doi: 10.3389/fphar.2018.00347. - DOI - PMC - PubMed
    1. Zeng H., Wang Y., Gu Y., Wang J., Zhang H., Gao H., Jin Q., Zhao L. Polydatin attenuates reactive oxygen species-induced airway remodeling by promoting Nrf2-mediated antioxidant signaling in asthma mouse model. Life Sci. 2019;218:25–30. doi: 10.1016/j.lfs.2018.08.013. - DOI - PubMed
    1. Ding W., Dong M., Deng J., Yan D., Liu Y., Xu T., Liu J. Polydatin attenuates cardiac hypertrophy through modulation of cardiac Ca2+ handling and calcineurin-NFAT signaling pathway. Am. J. Physiol. Heart Circ. Physiol. 2014;307:H792–H802. doi: 10.1152/ajpheart.00017.2014. - DOI - PubMed

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