A study of cytotoxic effect of the uremic toxin indoxyl sulfate on myoblasts in vitro, the expression of myostatin mRNA in myoblast cell culture, and the possibility of exogenous regulation

Introduction: indoxyl sulfate is a uremic toxin considered as one of the sources of muscle failure (cachexia) in patients with renal failure; this category of patients has a poor prognosis. Objective: an examination of the possibilities of exogenous regulation of the influence of indoxyl sulfate on myoblasts in vitro. Material and methods: the culture of primary mouse myoblasts was obtained according to the standard method. The cells were cultured in DMEM medium with 10% fetal calf serum (FST) (Gibco) at 37 °C and 6% CO2. The study of the effect of indoxyl sulfate on the proliferation rate of myoblasts was carried out by measuring the rate of reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma). To measure the expression of myostatin, RNA was isolated from mouse myoblasts using the Trizol reagent (Thermo Fisher Scientific). Reverse transcription was performed using a set of reagents from Silex. To compare the expression of myostatin mRNA in myoblasts, a real-time polymerase chain reaction was used. Ethylmethylhydroxypyridine malate was used at a physiological concentration of 70 μg/ml. Results: we have shown that indoxyl sulfate has a significant cytostatic effect on myoblasts in vitro, suppressing their proliferation. Ethyl methyl hydroxypyridine malate showed a statistically significant protective effect for myoblasts treated with the toxicant. In experiments where the expression of myostatin mRNA was studied, indoxyl sulfate did activate the expression for euther 48 hours or 72 hours. However, the expression of myostatin mRNA increased with the combined use of indoxyl sulfate and ethylmethylhydroxypyridine malate for 48 hours and persisted after 72 hours. Conclusion: ethylmethylhydroxypyridine malate, leveling the toxic effect of indoxyl sulfate, has a protective effect on myoblasts in vitro. The mechanism of joint activation of mRNA expression of myostatin indoxyl sulfate and ethylmethylhydroxypyridine malate remains unclear and requires further study.

индоксилсульфат, хроническая почечная недостаточность, миостатин, этилметилгидроксипиридина малат (этоксидол), indoxyl sulfate, chronic renal failure, myostatin, ethylmethylhydroxypyridine malate (ethoxydol)

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