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Метод неинвазивной оценки влияния биологически активных веществ на скорость восстановления уровня рН в мышце после предельной нагрузки с помощью 1Н МРС

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Цель исследования: с помощью протонной магнитно-резонансной спектроскопии (1Н МРС) определить возможность неинвазивной оценки влияния креатина и бета-аланина на скорость восстановления уровня рН в мышце после предельной нагрузки.Материал и методы. В качестве тестируемых биологически активных веществ (БАВ) были взяты креатина моногидрат и бета-аланин, применяемые согласно рекомендациям производителя. На первом этапе были построены калибровочные кривые зависимости рН от величины химического сдвига при снятии 1Н спектров модельных растворов дипептида карнозина для неинвазивного определения внутримышечного рН. Далее проводились эксперименты на лабораторных животных (мыши линии BALB/c) с использованием 9 Тл ЯМР спектрометра Bruker Advance III WB 400 МГц WB (Bruker, Germany). Следующим шагом стали эксперименты на добровольцах по отработке методики оценки эффектов БАВ на прямой четырехглавой мышце бедра. Использовался функциональный тест pwc170 в варианте степ-теста, позволяющий достичь закисления цитоплазмы миоцитов лактатом и оценить эффективность исследуемых БАВ на запас выносливости и функциональность аэробных систем по скорости восстановления уровня рН исследуемой мышцы. Дальнейшее сканирование осуществлялось с помощью высокопольного магнитно-резонансного томографа (Philips Healthcare, Achieva 3.0 Tл, North Braband, the Nederlands) и двух поверхностных кольцевидных радиочастотных катушек SENSE Flex-L.Результаты. Методом 1Н МРС было оценено влияние перорального приема креатина и бета-аланина на восстановление рН прямой четырехглавой мышцы бедра после закисления цитоплазмы миоцитов лактатом. Эксперименты с участием мелких лабораторных животных показали необходимость разработки и использования более точных методик выделения вокселя и подавления сигнала от жировой ткани при доклинической in vivo 1H спектроскопии для надежной фиксации химических сдвигов пиков карнозина. С помощью полученных для добровольцев протоколов удалось достичь воспроизводимости результатов, оптимальных соотношения сигнал/шум и ширины спектральных пиков карнозина с помощью 1H МРС при 3 Tл в прямой четырехглавой мышце бедра.Заключение. Полученные с помощью 1Н МРС данные на добровольцах позволяют сделать вывод, что разработанная методика дает возможность неинвазивной оценки влияния БАВ на скорость восстановления уровня рН в мышце после предельной нагрузки в режиме реального времени in vivo.

Ключевые слова:
магнитно-резонансная томография, рН-буферизация, карнозин, БАД, magnetic resonance imaging, рН-buffering, carnosine, BAS

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Method for non-invasive assessment of the effect of biologically active substances on the rate of the pH level restoration in the muscle after maximum load using 1H MRS

Purpose of the study: To determine the possibility of a non-invasive evaluation of the biologically active substances (BAS) effect on the rate of a pH level restoration in a muscle after a maximum load using 1H magnetic resonance spectroscopy (MRS).Materials and methods. Creatine monohydrate and beta-alanine were taken as tested biologically active substances, used according to the manufacturer's recommendations. At the first stage, calibration curves of a pH dependence on the magnitude of chemical shifts were plotted during assigning 1H spectra of model carnosine dipeptide solutions for non-invasive determination of intramuscular pH. Further experiments were carried out on laboratory animals (mice) using a 9 T NMR spectrometer Bruker Advance III WB 400MHz WB (Bruker, Germany). In experiments on volunteers the functional test pwc170 was used for assessing the ergogenic effects of biologically active substances on rectus quadriceps femoris. The test allows to achieve the level of myocytes cytoplasm acidification with lactate, and the effectiveness of functional biologically active substances on endurance, and also the function of aerobic systems by the muscle pH rate of recovery. Detection was performed using a high-field magnetic resonance imaging scanner (Philips Healthcare, Achieva 3.0T, North Braband, The Netherlands) and two SENSE Flex-L surface ring radiofrequency coils.Results. The effect of oral intake of creatine and beta-alanine on the restoration of rectus quadriceps femoris muscle pH after an acidification of the myocytes cytoplasm with lactate was evaluated using the 1H MRS method. Reproducible results with optimal signal-to-noise ratios and width of carnosine spectral peaks were achieved in volunteers using individual protocols and 1H MRS at 3T in the quadriceps femoris. Animal experiments have highlighted the need to develop and use more accurate techniques for voxel extraction and fat suppression during in vivo 1H spectroscopy to reliably capture the chemical shifts of carnosine peaks.Conclusion. The data obtained using 1H MRS on volunteers allow us to conclude that the developed method makes it possible to non-invasively assess the effect of biologically active substances on the rate of restoration of pH level in a muscle after a critical load in real time in vivo.

Keywords:
магнитно-резонансная томография, рН-буферизация, карнозин, БАД, magnetic resonance imaging, рН-buffering, carnosine, BAS