CRISPR/Cas9-mediated MSTN gene editing induced mitochondrial alterations in C2C12 myoblast cells

Background: Myostatin (MSTN) negatively regulates muscle mass and is a potent regulator of energy metabolism. However, MSTN knockout have affect mitochondrial function. This research assessed the mitochondrial energy metabolism of Mstn−/+ KO cells, and wondered whether the mitochondria biogenesis are affected. Results: In this study, we successfully achieved Mstn knockout in skeletal muscle C2C12 cells using a CRISPR/Cas9 system and measured proliferation and differentiation using the Cell-Counting Kit-8 assay and qPCR, respectively. We found that MSTN dysfunction could promote proliferation and differentiation compared with the behaviour of wild-type cells. Moreover, Mstn KO induced an increase in KIF5B expression. The mitochondrial content was significantly increased in Mstn KO C2C12 cells, apparently associated with the increases in PGC-1α, Cox1, Cox2, ND1 and ND2 expression. However, no differences were observed in glucose consumption and lactate production. Interestingly, Mstn KO C2C12 cells showed an increase in IL6 and a decrease in TNF-1α levels. Conclusion: These findings indicate that MSTN regulates mitochondrial biogenesis and metabolism. This gene-editing cells provided favourable evidence for animal breeding and metabolic diseases.

Effects of carbohydrate restriction on exercise performance / 体力科学

<p>Carbohydrate is a crucial energy fuel for exercise, and carbohydrate supplementation as peri-exercise has beneficial effects on exercise performance. However, recent studies have indicated the possibility that periodized carbohydrate restriction improves exercise performance. Carbohydrate restriction before exercise increases fatty-acid oxidation (FAO) and alternatively prevents carbohydrate consumption during exercise. This may contribute to the prevention of muscle glycogen depletion during endurance exercise competition. Additionally, acute and chronic studies have shown that peri-exercise carbohydrate restriction enhances molecular and functional adaptation related to FAO. Similarly, exercise training in a low-muscle glycogen state accompanied by carbohydrate restriction was reported to enhance mitochondrial biogenesis and improve FAO capacity, aerobic capacity, and endurance performance in untrained and highly trained subjects. The potential mechanism of these metabolic adaptations may be related to elevated circulating fatty-acid and adrenaline concentration during exercise with carbohydrate restriction and/or a low-muscle glycogen state. In addition, a decrease in muscle glycogen content may trigger signaling pathways related to FAO and mitochondria biogenesis by activating proteins with a glycogen-binding domain. This article reviews the effects of exercise with carbohydrate restriction and/or low-muscle glycogen state on metabolic adaptation and exercise performance and describes the molecular mechanisms and availability.</p>

Endogenous nitric oxide synthase inhibitor increase skeletal muscle con-tractility and mitochondria biosynthesis in 4-week running rats / 中国病理生理杂志

[ ABSTRACT] AIM:To observe the effect of endogenous nitric oxide synthase ( NOS) inhibitor asymmetric dimeth-ylarginine ( ADMA ) and its signaling pathways on NO levels and skeletal muscle contractility in 4-week running rats. METHODS:The 4 weeks running rat model was established.The twitch tension, tetanic tension and the fatigue test of sole-us muscle induced by electrical stimulation ex vivo were detected.The ATP content, mitochondrial DNA levels and the mR-NA expression of peroxisome proliferator-activated receptor γcoactivator-1α(PGC-1α), nuclear respiratory factor (NRF) were measured to reflect the mitochondrial function and biosynthesis in the skeletal muscle.Serum ADMA concentration was detected by high performance liquid chromatography.The endogenous ADMA enzymes PRMT1 and 2 subtypes of ADMA me-tabolism enzyme DDAH, 3 subtypes of NOS protein expression in the skeletal muscle were determined by Western blot.NOS activity and nitric oxide ( NO) content were analyzed by colorimetric method.RESULTS: Compared with normal control group, the twitch tension, tetanic tension and the anti-fatigue capability of soleus muscle in running group were significantly enhanced, ATP content, mitochondrial DNA content and the mRNA expression of PGC-1αand NRF were significantly in-creased (P ance of soleus muscle.The mechanism may be that increased cNOS expression feedbacks to increase ADMA concentration, thus maintaining the increase in NO synthesis at a relatively low level, and resulting in promoting skeletal muscle mitochon-dria biosynthesis and mitochondrial function.