Abstract

Exogenous lactate administration has more recently been investigated for its various prophylactic effects. Lactate derived from potential functional foods, such as fermented oyster extract (FO), may emerge as a practical and effective method of consuming exogenous lactate. The current study endeavored to ascertain whether the lactate derived from FO may act on muscle cell biology, and to what extent this may translate into physical fitness improvements. We examined the effects of FO in vitro and in vivo, on mouse C2C12 cells and exercise performance indicators in mice, respectively. In vitro, biochemical analysis was carried out to determine the effects of FO on lactate content and muscle cell energy metabolism, including adenosine triphosphate (ATP) activity. Western blot analysis was also utilized to measure the protein expression of total adenosine monophosphate-activated protein kinase (AMPK), p-AMPK (Thr172), lactate dehydrogenase (LDH), succinate dehydrogenase (SDHA) and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) in response to FO administration. Three experimental groups were formed: a positive control (PC) treated with 1% horse serum, FO10 treated with 10 μg/mL and FO50 treated with 50 μg/mL. In vivo, the effects of FO supplementation on exercise endurance were measured using the Rota-rod test, and Western blot analysis measured myosin heavy-chain 2 (MYH2) to assess skeletal muscle growth, alongside p-AMPK, total-AMPK, PGC-1α, cytochrome C and UCP3 protein expression. Biochemical analysis was also performed on muscle tissue to measure the changes in concentration of liver lactate, lactate dehydrogenase (LDH), glycogen and citrate. Five groups (n = 10/per group) consisted of a control group (CON), exercise group (Ex), positive control treated with Ex and 500 mg/kg Taurine (Ex-Tau), Ex and 100 mg/kg FO supplementation (Ex-FO100) and Ex and 200 mg/kg FO supplementation (Ex-FO200) orally administered over the 4-week experimental period.FO50 significantly increased PGC-1α expression (p < 0.001), whereas both FO10 and FO50 increased the expression of p-AMPK (p < 0.001), in C2C12 muscle cells, showing increased signaling important for mitochondrial metabolism and biogenesis. Muscle lactate levels were also significantly increased following FO10 (p < 0.05) and FO50 (p < 0.001). In vivo, muscle protein expression of p-AMPK (p < 0.05) and PGC-1α were increased, corroborating our in vitro results. Cytochrome C also significantly increased following FO200 intake. These results suggest that the effects of FO supplementation may manifest in a dose-response manner. FO administration, in vitro, and supplementation, in vivo, both demonstrate a potential for improvements in mitochondrial metabolism and biogenesis, and even for potentiating the adaptive effects of endurance exercise. Mechanistically, lactate may be an important molecule in explaining the aforementioned positive effects of FO.

Highlights

  • Regular physical activity is an established approach to improved quality of life and a preventative measure in the case of chronic and/or degenerative diseases

  • Biochemical and Western blot analysis: Biochemical analysis of myoblast C2C12 cells administered administered with FO10 and FO50 demonstrated a significant increase in lactate concentration in a with FO10 and FO50 demonstrated a significant increase in lactate concentration in a dose-response dose-response manner, p < 0.05 and p < 0.001, respectively (Figure 4)

  • The results of the present study demonstrate that, in vitro, mitochondrial homeostasis regulator proliferator-activated receptor gamma coactivator-1α (PGC-1α) was significantly upregulated by FO50 administration alongside the upregulation of upstream p-AMPK required for the phosphorylation of PGC-1α (p < 0.001)

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Summary

Introduction

Regular physical activity is an established approach to improved quality of life and a preventative measure in the case of chronic and/or degenerative diseases. Exercise approaches, such as endurance training, have been shown to induce a number of advantageous physiological adaptations on skeletal muscle (SkM), including remodeling of the mitochondrial network, the contractile apparatus and the vasculatures [1,2], which underlie many of the health benefits. The more recent lactate paradigm suggests that it counteracts acidosis, maintains neuron and astrocyte function [3] and fuels aerobic metabolism It is an important intermediary in numerous metabolic processes [4]. Mitochondrial homeostasis is heavily regulated by peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), a transcriptional coactivator central to the upregulation of mitochondrial biogenesis in response to exercise [10,11]

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