scbmtБИОМЕДИЦИНАJournal Biomed2074-59822713-0428Scientific center of biomedical technologies of Federal Medical and Biological Agency10.33647/2074-5982-18-2-17-30scbmt-1384Research ArticleМЕТОДЫ И ТЕХНОЛОГИИ БИОМЕДИЦИНСКИХ ИССЛЕДОВАНИЙMETHODS AND TECHNOLOGIES OF BIOMEDICAL RESEARCHДинамика стабильности экспрессии генов sdha, hprt, prl3d1 и hes1 в рамках моделирования фиброза печени крысStability of the sdha, hprt, prl3d1 and hes1 Gene Expression in a Rat Liver Fibrosis ModelЛебедеваЕ. И.LebedevaE. I.

 к.б.н., доц., 

 210009, Республика Беларусь, Витебск, просп. Фрунзе, 27 

 Cand. Sci. (Biol.), Assoc. Prof., 

 210009, Republic of Belarus, Vitebsk, Frunze Ave., 27 

lebedeva.ya-elenale2013@yandex.ruБабенкоА. С.BabenkoA. S.

 к.х.н., доц., 

 220116, Республика Беларусь, Минск, просп. Дзержинского, 83 

 Cand. Sci. (Chem.), Assoc. Prof.,

 220116, Republic of Belarus, Minsk, Dzerzhinsky Ave., 83, build. 15 

ЩастныйА. Т.ShchastniyA. T.

 д.м.н., проф.,

 210009, Республика Беларусь, Витебск, просп. Фрунзе, 27 

 Dr. Sci. (Med.), Prof.,

 210009, Republic of Belarus, Vitebsk, Frunze Ave., 27 

admin@vsmu.byУчреждение образования «Витебский государственный ордена Дружбы народов медицинский университет»БеларусьVitebsk State Order of Peoples’ Friendship Medical University BelarusУчреждение образования «Белорусский государственный медицинский университет»БеларусьBelarussian State Medical UniversityBelarus2022100620221821730Copyright © Лебедева Е.И., Бабенко А.С., Щастный А.Т., 20222022Лебедева Е.И., Бабенко А.С., Щастный А.Т.Lebedeva E.I., Babenko A.S., Shchastniy A.T.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.scbmt.ru/jour/article/view/1384

До настоящего времени не удалось выявить универсального набора референсных генов для нормализации данных полимеразной цепной реакции в режиме реального времени. Многочисленные исследования, затрагивающие выбор референсных генов для конкретных целей, далеко не всегда тщательно прорабатывают стратегию выбора. В ряде работ этап выбора референсных генов в принципе не используется ввиду дороговизны и иных причин. В результате этого для нормализации данных часто используются гены, неплохо зарекомендовавшие себя в иных, часто отличающихся, обстоятельствах эксперимента. Цель настоящего исследования заключалась в изучении динамики уровня мРНК генов в рамках моделирования фиброза печени крыс тиоацетамидом.При рассмотрении процесса фиброгенеза в целом, от интактной печени до развитого в ней фиброза, оптимальными референсными генами являются hes1 и sdha. Однако при акцентах на конкретных этапах фиброза стоит выбирать пару генов в зависимости от показателей стабильности. На начальных стадиях фиброгенеза можно использовать sdha и hprt. Ген hes1 наилучшим образом подойдёт на роль референсного в том случае, когда среднее значение Cq генов-мишеней будет составлять примерно 29 циклов (как у hes1). С осторожностью стоит использовать hes1 при работе в диапазонах Cq генов-мишеней 26–29 и более 30, поскольку ошибка в таком случае будет нарастать. В отношение гена sdha, придерживаясь того же принципа, отметим как оптимум значение Cq, равное 27. В то же время допустима работа с диапазоном Cq 24–27, а в диапазонах выше 28 использование sdha может быть сопряжено с повышением ошибок расчётов.

So far, no versatile set of reference genes for normalizing real-time polymerase chain reaction data has been identified. Numerous studies focusing the selection of reference genes for specific purposes frequently fail to elaborate a suitable selection strategy. In a number of such studies, the stage of selecting reference genes is ignored due to either its high cost or other reasons. As a result, the normalization of data is carried out using genes, which have previously shown their effectiveness under other, sometimes completely different, experimental conditions. In this work, we aim to study variations in the level of mRNA expression of several genes, some of which are commonly used to normalize RT-PCR data. As special conditions, modeling of rat liver fibrosis with thioacetamide was used.In our experiment, when considering the process of fibrogenesis as a whole, the optimal reference genes were found to be hes1 and sdha. However, when focusing on specific stages of fibrosis, a pair of genes should be selected depending on the stability indicators. At the initial fibrogenesis stages, sdha and hprt can be used. The hes1 gene is suitable as a reference gene, when the average Cq value of the target genes is approximately 29 cycles (as in hes1). Hes1 should be used with care when working in the Cq ranges of target genes of 26–29 and above 30, since the error is likely to increase. Following the same principle, the optimum Cq value for the sdha gene was observed to be 27, although the Cq range of 24–27 is also acceptable. At the same time, when working in the Cq range of above 28, the use of sdha may be associated with an increase in calculation errors.

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The authors declare that there are no conflicts of interest present.