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Advanced glycation end products promote ROS production via PKC/p47 phox axis in skeletal muscle cells

The Journal of Physiological Sciences volume 74, Article number: 51 (2024) Cite this article

Abstract

Advanced glycation end products (AGEs) are risk factors for various diseases, including sarcopenia. One of the deleterious effects of AGEs is the induction of abnormal reactive oxygen species (ROS) production in skeletal muscle. However, the underlying mechanism remains poorly understood. Therefore, the aim of this study was to elucidate how AGEs induce ROS production in skeletal muscle cells. This study demonstrated that AGEs treatment promoted ROS production in myoblasts and myotubes while PKC inhibitor abolished ROS production by AGEs stimulation. Phosphorylation of p47 phox by kinases such as PKCα is required to form the Nox2 complex, which induces ROS production. In this study, AGEs treatment promoted the phosphorylation of PKCα and p47 phox in myoblasts and myotubes. Our findings suggest that AGEs promote ROS production through the phosphorylation of PKCα and p47 phox in skeletal muscle cells.

Introduction

Type 2 diabetes mellitus (T2DM) is a significant public health issue. Previous studies estimated that 382 million individuals had T2DM in 2013 and that this number will exceed 590 million people by 2035 worldwide [1, 2]. This indicates a rapid increase in the number of T2DM patients. T2DM is associated with mortality and morbidity, and causes multiple adverse conditions including insulin resistance and hyperglycemia [3]. Previous studies have reported that advanced glycation end products (AGEs) are accumulated in T2DM patients [4, 5]. AGEs are formed by a nonenzymatic glycation reaction known as the Maillard reaction [6], and have deleterious effects on various tissues. For instance, AGEs are recognized as risk factors for Alzheimer’s disease and sarcopenia [7, 8]. Since AGEs induce various signal pathways associated with the pathophysiology of diabetic complications [9], this study aimed to elucidate how AGEs play a deleterious role.

One of the adverse effects of AGEs is the induction of reactive oxygen species (ROS) production. Previous reports have demonstrated that AGEs induce ROS production in various cell types [10,11,12,13]. In particular, it is suggested that AGEs promote ROS production via NADPH oxidase 2 (Nox2) activation in renal glomerular tissue and hepatic stellate cells [13, 14]. The Nox2 complex is composed of gp91 phox, p22 phox, p67 phox, p47 phox, p40 phox and Rac. To activate Nox2, p47 phox is required to be phosphorylated. Protein kinase C (PKC) is phosphorylated and activated by various factors, including growth factors and hyperglycemia [15,16,17]. PKC phosphorylates p47 phox, activates Nox2 and induces ROS production [18].

AGEs impair the functions of skeletal muscle [19], which is the most abundant tissue in the body and has fundamental multiple functions such as movement, heat production and metabolism [20, 21]. The dysfunction and atrophy of skeletal muscle can cause several detrimental conditions and shorter life expectancy [22]. AGEs cause insulin resistance by blunting the insulin sensitivity of skeletal muscle [23], and promote ROS production in skeletal muscle tissue [24]. Excessive ROS levels impair skeletal muscle function and damage the skeletal muscle cells [25], necessitating the appropriate neutralization of ROS by antioxidant enzymes such as catalase.

While previous report has demonstrated that AGEs promote ROS production in skeletal muscle tissue [24], the signal pathway of ROS production by AGEs remains to be elucidated. In hepatic stellate cells, AGEs induce ROS production via Nox2, which is activated by PKC [13]. Therefore, we hypothesized that AGEs activate PKC and Nox2, and then promote ROS production in skeletal muscle cells.

Materials and methods

Cell culture

Mouse myoblast C2C12 cells were cultured in growth medium containing Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Biowest, Nuaillé, France) on 35 mm culture dishes (Asone, Osaka, Japan), 96 well clear bottom plates (TPP Techno Plastic Products AG, Trasadingen, Switzerland) or 96 well black bottom plates (Sumitomo Bakelite, Tokyo, Japan), which are coated with type I collagen (Corning Incorporated, Corning, NY, USA) at 37 °C in a 5% CO2 incubator. To differentiate C2C12 myoblasts into myotubes, the culture medium was changed to differential medium containing DMEM supplemented with 2% heat-inactivated horse serum (HS; Thermo Fisher Scientific) 2 days after myoblasts were cultured with growth medium. In this study, we used myotubes 6 days after differentiation.

AGEs stimulation

AGEs were prepared as described previously [26]. Briefly, 50 mg/mL bovine serum albumin (BSA; Nacalai Tesque, Kyoto, Japan) was incubated with 0.1 M glyceraldehyde (Sigma-Aldrich Co., St. Louis, MO, USA) in 0.2 M phosphate buffer (pH, 7.4) under sterile conditions at 37 °C for 7 days. Unincorporated glyceraldehyde was removed by dialysis. Non-glycated BSA was incubated under the same conditions except in the absence of glyceraldehyde as a negative control.

Myoblasts were treated with 1 mg/ml AGEs or 1 mg/ml BSA for 1 h while myotubes were treated with 1 mg/ml AGEs or 1 mg/ml BSA for 24 h. To inhibit PKC activity, both myoblasts and myotubes were incubated with 100 nM PKC antagonist Go6983 (Abcam, Cambridge, UK) for 30 min prior to AGEs stimulation. These cells were used for subsequent experiments.

ROS production analysis

To analyze ROS production, the Highly Sensitive DCFH-DA ROS Assay kit (Dojindo Laboratories, Kumamoto, Japan) was used according to the manufacture’s instruction. Briefly, cells were washed with Hanks' Balanced Salts Solution (HBSS; Thermo Fisher Scientific) after AGEs stimulation, and then incubated with DCFH-DA solution for 30 min. After washing with HBSS, ROS production was analyzed by microscopy or plate reader. For microscopic observation, images were captured at × 10 magnification using BZ-X800 (Keyence, Osaka, Japan). To analyze fluorescent intensity, Spark (Tecan, Männedorf, Switzerland) was used at excitation/emission: 490/540 nm.

Western blotting

After AGEs stimulation, cells were lysed with radioimmunoprecipitation assay buffer (RIPA buffer) containing with 50 mM Tris, 3 mM MgCl2, 100 mM NaCl, 1 mM dithiothreitol (DTT), 1% triton X-100, 0.1% sodium dodecyl sulfate (SDS), 0.1% sodium deoxycholate, 1 mM phenylmethylsulfonyl fluoride (PMSF), 10 μM leupeptin, 10 mM NaF and 10 mM b-glycerophosphate. After centrifugation at 17,000 × g for 10 min at 4 °C, the supernatant was diluted with an equal volume of 2 × Laemmli sample buffer containing with 4% SDS, 200 mM DTT, 100 mM Tris, 20% glycerol and 0.05% bromophenol blue, and then boiled at 95 °C for 5 min. The samples were subjected to SDS-PAGE and transferred to Immobilon-P Transfer Membrane (Merck, Darmstadt, Germany). The membrane was blocked with PVDF blocking reagent (Toyobo, Osaka, Japan) for 1 h at room temperature and then incubated with anti-phospho-PKCα antibody (1:1000, sc-377565, Santa Cruz Biotechnology, Dallas, Texas, USA) diluted with TBST or anti-phospho-p47 phox antibody (1:5000, SAB4504289, Sigma-Aldrich Co.) diluted with Can Get Signal Solution 1 (Toyobo) overnight at 4 °C. Then, the membrane was incubated with anti-mouse IgG conjugated with horseradish peroxidase (HRP) antibody (1:5000, NA931V, Cytiva, Tokyo, Japan) or anti-rabbit IgG conjugated with HRP antibody (1:5000, NA934V, Cytiva) diluted with Can Get Signal Solution 2 (Toyobo). The membrane was treated with Chemi-Lumi One Ultra (Nacalai Tesque) and captured by LuminoGraph (Atto, Tokyo, Japan). To reprobe antibodies, the membrane was incubated with 15% H2O2 for 30 min at room temperature or WB Stripping Solution Strong (Nacalai Tesque) for 10 min at room temperature. Similarly, the membrane was blocked and incubated with anti-PKCα antibody (1:1000, sc-8393, Santa Cruz Biotechnology) diluted with TBST or anti-p47 phox antibody (1:1000, sc-17845, Santa Cruz Biotechnology) diluted with Can Get Signal Solution 1, incubated with appropriate secondary antibodies and then treated with Chemi-Limi One Ultra. After detection with LuminoGraph, the membrane was stained with Coomassie Brilliant Blue (CBB) solution containing with 40% MeOH, 10% AcOH and 0.05% CBB R-250 (Nacalai Tesque). Signal intensity was measured by ImageJ (National Institute of Health, Bethesda, MD, USA). The signal intensity of the target protein was normalized to total protein (CBB staining intensity).

Statistics

All values were presented as means ± standard error of the mean (SEM). Statistical analyses were performed using GraphPad Prism 10 (GraphPad Software, Boston, MA, USA). To compare the means between the two groups, Mann–Whitney U test was used. Two-way ANOVA with Tukey’s multiple comparison test was employed for comparisons of the mean among the four groups. Differences among groups were considered significant at p < 0.05.

Results

AGEs promote ROS production in myoblasts

To examine the effect of AGEs on ROS production in myoblasts, C2C12 cells were treated with 1 mg/ml AGEs or 1 mg/ml BSA for 1 h. BSA was used as control for AGEs. After the cells were treated with DCFH-DH, ROS production was measured. Consequently, AGEs treatment significantly increased the fluorescent intensity compared to BSA treatment, suggesting that AGEs promote ROS production in myoblasts (Fig. 1).

Fig. 1
figure 1

Advanced glycation end products (AGEs) promote radical oxygen species (ROS) production in C2C12 myoblasts. Myoblasts are treated with 1 mg/ml AGEs or 1 mg/ml BSA for 1 h. A The representative image of myoblasts stained with DCFH-DA. Scale bars, 200 μm. B Quantification of fluorescent intensity of myoblasts stained with DCFH-DA (n = 16). Error bars show mean ± SEM. ** indicates p < 0.01 by Mann–Whitney U test

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AGEs induce phosphorylation of PKCα and p47 phox in myoblasts

Because previous studies reported that PKCα induces phosphorylation of p47 phox, activates Nox2, and then promotes ROS production [14, 18, 27], we hypothesized that AGEs induce the phosphorylation of PKCα and p47 phox, and then promote ROS production in myoblasts.

To verify this hypothesis, myoblasts were stimulated with AGEs as described above, and the amount of p-PKCα and p-p47 phox was measured. While the ratio of p-PKCα to PKCα was tended to be increased, p-PKCα was significantly increased by AGEs stimulation (Fig. 2A–C). While the ratio of p-p47 phox to p47 phox was significantly increased, p-p47 phox was tended to be increased by AGEs treatment (Fig. 2D–F). Taken together, it is suggested that AGEs promote the phosphorylation of both PKCα and p47 phox in myoblasts.

Fig. 2
figure 2

AGEs promote phosphorylation of PKCα and p47 phox in C2C12 myoblasts. Myoblasts are treated with 1 mg/ml AGEs or 1 mg/ml BSA for 1 h. A The representative immunoblot image of p-PKCα and PKCα. B–C Quantification of the expression level of p-PKCα normalized to PKCα or total protein (n = 12). D The representative immunoblot image of p-p47 phox and p47 phox. E–F Quantification of the expression level of p-p47 phox normalized to p47 phox or total protein (n = 12). For all graphs, error bars show mean ± SEM. * indicates p < 0.05 by Mann–Whitney U test

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ROS production is induced by AGEs in myotubes

To analyze the effect of AGEs on ROS induction in myotubes, myotubes were treated with 1 mg/ml AGEs or 1 mg/ml BSA for 24 h. After DCFH-DA treatment, the fluorescent intensity was significantly increased by AGEs treatment, indicating that AGEs treatment promotes ROS production in myotubes (Fig. 3).

Fig. 3
figure 3

AGEs promote ROS production in C2C12 myotubes. Myotubes are treated with 1 mg/ml AGEs or 1 mg/ml BSA for 24 h. A The representative image of myotubes stained with DCFH-DA. Scale bars, 200 μm. B Quantification of fluorescent intensity of myotubes stained with DCFH-DA (n = 16). Error bars show mean ± SEM. ** indicates p < 0.01 by Mann–Whitney U test

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Phosphorylation of PKCα and p47 phox is promoted by AGEs in myotubes

To examine whether AGEs promote the phosphorylation of PKCα and p47 phox in myotubes, myotubes were treated with AGEs in a similar manner as described above. While the ratio of p-PKCα to PKCα was tended to be increased, p-PKCα was significantly increased by AGEs treatment (Fig. 4A–C). AGEs stimulation significantly increased both p-p47 phox and the ratio of p-p47 phox to p47 phox (Fig. 4D–F). Collectively, it is suggested that AGEs induce the phosphorylation of both PKCα and p47 phox in myotubes.

Fig. 4
figure 4

AGEs promote phosphorylation of PKCα and p47 phox in C2C12 myotubes. Myotubes are treated with 1 mg/ml AGEs or 1 mg/ml BSA for 24 h. A The representative immunoblot image of p-PKCα and PKCα. B–C Quantification of the expression level of p-PKCα normalized to PKCα or total protein (n = 15). D The representative immunoblot image of p-p47 phox and p47 phox. E–F Quantification of the expression level of p-p47 phox normalized to p47 phox or total protein (n = 15). For all graphs, error bars show mean ± SEM. * and **** indicate p < 0.05 and p < 0.0001, respectively, by Mann–Whitney U test

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PKC antagonist abolishes ROS induction by AGEs in myoblasts and myotubes

To confirm whether AGEs promote ROS production via PKC, cells were treated with 100 nM PKC antagonist Go6983 for 30 min prior to AGEs stimulation. AGEs stimulation induced ROS production in both myoblasts and myotubes whereas Go6983 abolished ROS induction by AGEs, suggesting that AGEs induce ROS production via PKC (Fig. 5).

Fig.5
figure 5

PKC inhibitor abrogates ROS induction by AGEs treatment in C2C12 myoblasts and myotubes. Following a 30 min preincubation with 100 nM Go6983, myoblasts are treated with 1 mg/ml AGEs or 1 mg/ml BSA for 1 h while myotubes are treated with 1 mg/ml AGEs or 1 mg/ml BSA for 24 h. A Quantification of fluorescent intensity of myoblasts (n = 16) and B myotubes stained with DCFH-DA (n = 16–32). For all graphs, error bars show mean ± SEM. ns, *** and **** indicate p > 0.05, p < 0.001 and p < 0.0001, respectively, by two-way ANOVA with Tuker’s multiple comparison test

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Discussion

This study investigated how AGEs induce ROS production in skeletal muscle cells. We showed that AGEs treatment increased the fluorescent intensity of DCFH-DA staining in myoblasts and myotubes, suggesting that AGEs promote ROS production (Figs. 1, 3). AGEs treatment also promoted the phosphorylation of PKCα and p47 phox in myoblasts and myotubes (Figs. 2, 4). Considering that PKC inhibitor abolished ROS production by AGEs stimulation (Fig. 5), we propose that AGEs induce ROS production through PKC/Nox2 axis in skeletal muscle.

In this study, AGEs promoted ROS production in myoblasts and myotubes in vitro (Figs. 1, 3). A previous report also suggests that ROS production is increased by AGEs treatment in skeletal muscle tissue in vivo [24]. However, the mechanism of ROS induction by AGEs in skeletal muscle is not fully understood. Because AGEs promote ROS production via Nox2 in hepatic stellate cells [13], we speculated that ROS production is induced by AGEs through Nox2. In this study, AGEs treatment promoted the phosphorylation of p47 phox, a component of the Nox2 complex, in myoblasts and myotubes (Figs. 2D–F, 4D–F). To produce ROS via Nox2, it is necessary to form the complex consisting of gp91 phox, p22 phox, p67 phox, p47 phox, p40 phox and Rac [18]. All components of the Nox2 complex are expressed in myoblasts and myotubes [28, 29]. Phosphorylation of p47 phox is required for the Nox2 complex formation. Considering that AGEs induce the phosphorylation of p47 phox (Figs. 2D–F, 4D–F), it is plausible that AGEs promote the ROS production via the phosphorylation of p47 phox in myoblasts and myotubes.

PKC is one of the kinases which phosphorylate p47 phox [27]. PKC belongs to a family of serine/threonine kinases and regulates a myriad of fundamental signal pathways, leading to the control of cell growth, differentiation, apoptosis and so on [30]. PKC is phosphorylated and activated by various ligands such as growth factors and Wnt [15, 31]. Since a previous report demonstrated that AGEs induce the phosphorylation of PKCα and ROS production in renal glomerular tissue [14], we assumed that AGEs also transduce the same signal pathway in skeletal muscle. Indeed, AGEs treatment promoted the phosphorylation of PKCα in myoblasts and myotubes (Figs. 2A–C, 4A–C). Taken together, this study suggests that PKCα is phosphorylated by AGEs treatment, and then PKCα phosphorylates p47 phox.

T2DM is a metabolic disorder which develops multifactorial symptoms including hyperglycemia. Hyperglycemia accelerates the formation of AGEs, which are involved in the pathogenesis of T2DM and diabetic complications [4, 9]. T2DM and AGEs cause multiple deleterious conditions such as abnormal ROS production [10,11,12, 32]. Excessive ROS burden impairs skeletal muscle function and causes frailty [33]. Considering that this study demonstrated that AGEs induce ROS production via Nox2 in skeletal muscle cells, Nox2 inhibitors may protect skeletal muscle from excessive ROS production. Indeed, gp91ds-tst, a peptide inhibitor of Nox2, mitigates skeletal muscle atrophy induced by hindlimb unloading [34].

The primary receptor of AGEs is the receptor for advanced glycation end products (RAGE). The interaction between AGEs and RAGE transduces various signal pathways, resulting in pathological conditions. Previous reports have demonstrated that AGE/RAGE signal activates PKC in cardiomyocytes [35] and induces ROS production via Nox2 in human umbilical vein endothelial cells (HUVEC) [11]. Together with this study, it is possible that AGEs interact with RAGE, phosphorylate PKC and p47 phox, and then induce ROS production.

We acknowledge that this study has several limitations. First, although this study demonstrated that AGEs promote the phosphorylation of PKCα and p47 phox in skeletal muscle cells (Figs. 2, 4), previous studies have suggested that AGEs induce ROS production by different PKC isoforms [10, 13, 14, 16] and these PKC isoforms phosphorylate p47 phox [27]. Therefore, it is possible that AGEs may phosphorylate p47 phox via activation of not only PKCα but also other PKC isoforms. Second, AGEs may promote ROS production via mitochondrial dysfunction as well as Nox2 activation in skeletal muscle cells. In Achilles tendon-derived fibroblasts, AGEs impair mitochondrial function [36]. Mitochondrial dysfunction causes abnormal ROS production [37]. Thus, further studies are needed to elucidate whether AGEs phosphorylate other PKC isoforms and whether AGEs induce ROS production via mitochondrial dysfunction.

Our results demonstrated that AGEs promoted ROS production in myoblasts and myotubes (Figs. 1, 3) while PKC inhibitor abolished ROS induction by AGEs (Fig. 5). p47 phox is a component of the Nox2 complex, which is one of the molecular systems associated with the induction of ROS production. AGEs promoted the phosphorylation of PKCα and p47 phox in myoblasts and myotubes (Figs. 2, 4). Therefore, this study suggests that AGEs-induced ROS production is mediated by PKC/p47 phox axis.

Data availability

The data underpinning the findings of this study are available from the corresponding author, TE, upon reasonable request.

References

  1. Guariguata L, Whiting DR, Hambleton I, Beagley J, Linnenkamp U, Shaw JE (2014) Global estimates of diabetes prevalence for 2013 and projections for 2035. Diabet Res Clin Pract 103:137–149. https://doi.org/10.1016/j.diabres.2013.11.002

    Article  CAS  Google Scholar 

  2. Ozougwu O (2013) The pathogenesis and pathophysiology of type 1 and type 2 diabetes mellitus. J Physiol Pathophysiol 4:46–57. https://doi.org/10.5897/jpap2013.0001

    Article  Google Scholar 

  3. Barr EL, Zimmet PZ, Welborn TA, Jolley D, Magliano DJ, Dunstan DW, Cameron AJ, Dwyer T, Taylor HR, Tonkin AM, Wong TY, McNeil J, Shaw JE (2007) Risk of cardiovascular and all-cause mortality in individuals with diabetes mellitus, impaired fasting glucose, and impaired glucose tolerance: the Australian diabetes, obesity, and lifestyle study (AusDiab). Circulation 116:151–157. https://doi.org/10.1161/CIRCULATIONAHA.106.685628

    Article  CAS  PubMed  Google Scholar 

  4. Nowotny K, Jung T, Hohn A, Weber D, Grune T (2015) Advanced glycation end products and oxidative stress in type 2 diabetes mellitus. Biomolecules 5:194–222. https://doi.org/10.3390/biom5010194

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Kilhovd BK, Berg TJ, Birkeland KI, Thorsby P, Hanssen KF (1999) Serum levels of advanced glycation end products are increased in patients with type 2 diabetes and coronary heart disease. Diabet Care 22:1543–1548. https://doi.org/10.2337/diacare.22.9.1543

    Article  CAS  Google Scholar 

  6. Maillard LC (1912) Action of amino acids on sugars. Formation of melanoidins in a methodical way. Compte Rendu De l’Academie Des Sci 154:66–68

    CAS  Google Scholar 

  7. Sasaki N, Fukatsu R, Tsuzuki K, Hayashi Y, Yoshida T, Fujii N, Koike T, Wakayama I, Yanagihara R, Garruto R, Amano N, Makita Z (1998) Advanced glycation end products in Alzheimer’s disease and other neurodegenerative diseases. Am J Pathol 153:1149–1155. https://doi.org/10.1016/S0002-9440(10)65659-3

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Yabuuchi J, Ueda S, Yamagishi SI, Nohara N, Nagasawa H, Wakabayashi K, Matsui T, Yuichiro H, Kadoguchi T, Otsuka T, Gohda T, Suzuki Y (2020) Association of advanced glycation end products with sarcopenia and frailty in chronic kidney disease. Sci Rep 10:17647. https://doi.org/10.1038/s41598-020-74673-x

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Khalid M, Petroianu G, Adem A (2022) Advanced glycation end products and diabetes mellitus: mechanisms and perspectives. Biomolecules. https://doi.org/10.3390/biom12040542

    Article  PubMed Central  PubMed  Google Scholar 

  10. Wang M, Zhang W, Xu S, Peng L, Wang Z, Liu H, Fang Q, Deng T, Men X, Lou J (2017) TRB3 mediates advanced glycation end product-induced apoptosis of pancreatic beta-cells through the protein kinase C beta pathway. Int J Mol Med 40:130–136. https://doi.org/10.3892/ijmm.2017.2991

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Wautier MP, Chappey O, Corda S, Stern DM, Schmidt AM, Wautier JL (2001) Activation of NADPH oxidase by AGE links oxidant stress to altered gene expression via RAGE. Am J Physiol Endocrinol Metab 280:E685-694. https://doi.org/10.1152/ajpendo.2001.280.5.E685

    Article  CAS  PubMed  Google Scholar 

  12. Ding Y, Kantarci A, Hasturk H, Trackman PC, Malabanan A, Van Dyke TE (2007) Activation of RAGE induces elevated O2- generation by mononuclear phagocytes in diabetes. J Leukoc Biol 81:520–527. https://doi.org/10.1189/jlb.0406262

    Article  CAS  PubMed  Google Scholar 

  13. Guimarães EL, Empsen C, Geerts A, van Grunsven LA (2010) Advanced glycation end products induce production of reactive oxygen species via the activation of NADPH oxidase in murine hepatic stellate cells. J Hepatol 52:389–397. https://doi.org/10.1016/j.jhep.2009.12.007

    Article  CAS  PubMed  Google Scholar 

  14. Thallas-Bonke V, Thorpe SR, Coughlan MT, Fukami K, Yap FY, Sourris KC, Penfold SA, Bach LA, Cooper ME, Forbes JM (2008) Inhibition of NADPH oxidase prevents advanced glycation end product-mediated damage in diabetic nephropathy through a protein kinase C-alpha-dependent pathway. Diabetes 57:460–469. https://doi.org/10.2337/db07-1119

    Article  CAS  PubMed  Google Scholar 

  15. Dowling CM, Kiely PA (2015) Targeting protein kinase C downstream of growth factor and adhesion signalling. Cancers 7:1271–1291. https://doi.org/10.3390/cancers7030836

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  16. Geraldes P, King GL (2010) Activation of protein kinase C isoforms and its impact on diabetic complications. Circ Res 106:1319–1331. https://doi.org/10.1161/CIRCRESAHA.110.217117

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Koya D, King GL (1998) Protein kinase C activation and the development of diabetic complications. Diabetes 47:859–866. https://doi.org/10.2337/diabetes.47.6.859

    Article  CAS  PubMed  Google Scholar 

  18. Vermot A, Petit-Hartlein I, Smith SME, Fieschi F (2021) NADPH oxidases (NOX): an overview from discovery molecular mechanisms to physiology and pathology. Antioxidants. https://doi.org/10.3390/antiox10060890

    Article  PubMed Central  PubMed  Google Scholar 

  19. Borchers A, Pieler T (2010) Programming pluripotent precursor cells derived from Xenopus embryos to generate specific tissues and organs. Genes 1:413–426. https://doi.org/10.3390/genes1030413

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. McKendry J, Currier BS, Lim C, McLeod JC, Thomas ACQ, Phillips SM (2020) Nutritional supplements to support resistance exercise in countering the sarcopenia of aging. Nutrients. https://doi.org/10.3390/nu12072057

    Article  PubMed Central  PubMed  Google Scholar 

  21. Yadav A, Yadav SS, Singh S, Dabur R (2022) Natural products: potential therapeutic agents to prevent skeletal muscle atrophy. Eur J Pharmacol 925:174995. https://doi.org/10.1016/j.ejphar.2022.174995

    Article  CAS  PubMed  Google Scholar 

  22. Kalyani RR, Corriere M, Ferrucci L (2014) Age-related and disease-related muscle loss: the effect of diabetes, obesity, and other diseases. Lancet Diabet Endocrinol 2:819–829. https://doi.org/10.1016/S2213-8587(14)70034-8

    Article  Google Scholar 

  23. Cassese A, Esposito I, Fiory F, Barbagallo AP, Paturzo F, Mirra P, Ulianich L, Giacco F, Iadicicco C, Lombardi A, Oriente F, Van Obberghen E, Beguinot F, Formisano P, Miele C (2008) In skeletal muscle advanced glycation end products (AGEs) inhibit insulin action and induce the formation of multimolecular complexes including the receptor for AGEs. J Biol Chem 283:36088–36099. https://doi.org/10.1074/jbc.M801698200

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Du H, Ma Y, Wang X, Zhang Y, Zhu L, Shi S, Pan S, Liu Z (2023) Advanced glycation end products induce skeletal muscle atrophy and insulin resistance via activating ROS-mediated ER stress PERK/FOXO1 signaling. Am J Physiol Endocrinol Metab 324:E279–E287. https://doi.org/10.1152/ajpendo.00218.2022

    Article  CAS  PubMed  Google Scholar 

  25. Zhang H, Qi G, Wang K, Yang J, Shen Y, Yang X, Chen X, Yao X, Gu X, Qi L, Zhou C, Sun H (2023) Oxidative stress: roles in skeletal muscle atrophy. Biochem Pharmacol 214:115664. https://doi.org/10.1016/j.bcp.2023.115664

    Article  CAS  PubMed  Google Scholar 

  26. Kong X, Wang GD, Ma MZ, Deng RY, Guo LQ, Zhang JX, Yang JR, Su Q (2015) Sesamin ameliorates advanced glycation end products-induced pancreatic beta-cell dysfunction and apoptosis. Nutrients 7:4689–4704. https://doi.org/10.3390/nu7064689

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Fontayne A, Dang PM, Gougerot-Pocidalo MA, El-Benna J (2002) Phosphorylation of p47phox sites by PKC alpha, beta II, delta, and zeta: effect on binding to p22phox and on NADPH oxidase activation. Biochemistry 41:7743–7750. https://doi.org/10.1021/bi011953s

    Article  CAS  PubMed  Google Scholar 

  28. Padron S, de Figueiredo A, Salmon AB, Bruno F, Jimenez F, Martinez HG, Halade GV, Ahuja SS, Clark RA, DeFronzo RA, Abboud HE, El Jamali A (2015) Nox2 mediates skeletal muscle insulin resistance induced by a high fat diet. J Biol Chem 290:13427–13439. https://doi.org/10.1074/jbc.M114.626077

    Article  CAS  Google Scholar 

  29. Charrasse S, Comunale F, Fortier M, Portales-Casamar E, Debant A, Gauthier-Rouviere C (2007) M-cadherin activates Rac1 GTPase through the Rho-GEF trio during myoblast fusion. Mol Biol Cell 18:1734–1743. https://doi.org/10.1091/mbc.e06-08-0766

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  30. Cosentino-Gomes D, Rocco-Machado N, Meyer-Fernandes JR (2012) Cell signaling through protein kinase C oxidation and activation. Int J Mol Sci 13:10697–10721. https://doi.org/10.3390/ijms130910697

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  31. Sheldahl LC, Park M, Malbon CC, Moon RT (1999) Protein kinase C is differentially stimulated by Wnt and Frizzled homologs in a G-protein-dependent manner. Current biology CB 9:695–698. https://doi.org/10.1016/s0960-9822(99)80310-8

    Article  CAS  PubMed  Google Scholar 

  32. Uryash A, Umlas J, Mijares A, Adams JA, Lopez JR (2023) Enhancing muscle intracellular Ca(2+) homeostasis and glucose uptake: passive pulsatile shear stress treatment in type 2 diabetes. Biomedicines. https://doi.org/10.3390/biomedicines11102596

    Article  PubMed Central  PubMed  Google Scholar 

  33. El Assar M, Angulo J, Rodriguez-Manas L (2020) Frailty as a phenotypic manifestation of underlying oxidative stress. Free Radic Biol Med 149:72–77. https://doi.org/10.1016/j.freeradbiomed.2019.08.011

    Article  CAS  PubMed  Google Scholar 

  34. Lawler JM, Hord JM, Ryan P, Holly D, Janini Gomes M, Rodriguez D, Guzzoni V, Garcia-Villatoro E, Green C, Lee Y, Little S, Garcia M, Hill L, Brooks MC, Lawler MS, Keys N, Mohajeri A, Kamal KY (2021) Nox2 inhibition regulates stress response and mitigates skeletal muscle fiber atrophy during simulated microgravity. Int J Mol Sci. https://doi.org/10.3390/ijms22063252

    Article  PubMed Central  PubMed  Google Scholar 

  35. Yu L, Zhao Y, Xu S, Ding F, Jin C, Fu G, Weng S (2013) Advanced glycation end product (AGE)-AGE receptor (RAGE) system upregulated connexin43 expression in rat cardiomyocytes via PKC and Erk MAPK pathways. Int J Mol Sci 14:2242–2257. https://doi.org/10.3390/ijms14022242

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  36. Patel SH, Yue F, Saw SK, Foguth R, Cannon JR, Shannahan JH, Kuang S, Sabbaghi A, Carroll CC (2019) Advanced glycation end-products suppress mitochondrial function and proliferative capacity of achilles tendon-derived fibroblasts. Sci Rep 9:12614. https://doi.org/10.1038/s41598-019-49062-8

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Bhatti JS, Bhatti GK, Reddy PH (2017) Mitochondrial dysfunction and oxidative stress in metabolic disorders—a step towards mitochondria based therapeutic strategies. Biochim Biophys Acta Mol Basis Dis 1863:1066–1077. https://doi.org/10.1016/j.bbadis.2016.11.010

    Article  CAS  PubMed  Google Scholar 

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Funding

This study was financially supported by JSPS KAKENHI Grant Numbers 21H03319, 22K18413 and 23H03283, and the Uehara Memorial Foundation.

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Authors and Affiliations

  1. Laboratory of Molecular Adaptations to Exercise, Graduate School of Human and Environmental Studies, Kyoto University, Yoshida-Nihonmatsu-Cho, Sakyo-Ku, Kyoto, 606-8501, Japan

    Shinichiro Suzuki & Tatsuro Egawa

  2. Laboratory of Sports and Exercise Medicine, Graduate School of Human and Environmental Studies, Kyoto University, Yoshida-Nihonmatsu-Cho, Sakyo-Ku, Kyoto, 606-8501, Japan

    Tatsuya Hayashi

Authors
  1. Shinichiro Suzuki
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  2. Tatsuya Hayashi
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  3. Tatsuro Egawa
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Contributions

SS performed and analyzed all experiments, and wrote the manuscript. TH and TE supervised the study. TE revised the manuscript.

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Correspondence to Tatsuro Egawa.

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Suzuki, S., Hayashi, T. & Egawa, T. Advanced glycation end products promote ROS production via PKC/p47 phox axis in skeletal muscle cells. J Physiol Sci 74, 51 (2024). https://doi.org/10.1186/s12576-024-00944-1

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  • DOI: https://doi.org/10.1186/s12576-024-00944-1

Keywords

The Journal of Physiological Sciences

ISSN: 1880-6562

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