Investigation of the effect of boiling on the level of milk miR-191

Year 2024, , 480 - 488, 28.09.2024

Fatih Atilla Bağcı , Dilek Pirim

https://doi.org/10.29050/harranziraat.1409969

Abstract

MicroRNAs (miRNAs) are small non-coding RNA sequences ~22 bp in length that play an active role in cellular processes. Recent studies have identified miRNA abundance in cow’s milk, highlighting their nutritional impact and their potential utilization as biomarkers of food quality. However, current research suggests that dietary intake of cow’s milk miRNAs may transfer to humans and have nutritional relevance for human health by entering human circulation and affecting important pathways associated with human diseases. Therefore, it is crucial to determine the miRNA content in milk and dairy products. The miR-191 has a similar sequence in cows and humans, and it has been previously shown to abundantly exist in cow milk. Here, we aimed to investigate the effects of boiling to the miR-191 levels in milk. Total RNA was isolated from raw and milk boiled at 100°C, and miR-191 levels in raw and boiled milk were analyzed by RT-qPCR method. Previous research reported that homogenization and pasteurization processes used in milk production stages have miRNA-specific distinct effects. After heat treatments, the amount of miR-191 was reduced by 95.8% (p<0.0001) in boiled raw milk and 66.4% (p=0.001) in boiled pasteurized milk compared to pasteurized milk. Meanwhile, we observed a statistically significant difference (p<0.0001) in the CT values obtained by quantification of miR-191 in raw and pasteurized milk. The results of our study present preliminary data for the effects of boiling milk on the milk miRNA content and point out the significance of miRNA-specific effects of milk processing steps on milk miRNA composition.

Keywords

miRNA, raw milk, boiled milk, pasteurized milk, RT-qPCR, miR-191

Project Number

FLO-2023-1551

Sütün kaynatılmasının süt miR-191 düzeyine etkisinin araştırılması

Abstract

MikroRNA'lar (miRNA'lar), gen anlatımının düzenlenmesinde etkin rol oynayan ~22 bp uzunluğunda küçük, kodlanmayan RNA dizileridir. Son yıllarda yapılan araştırmalarda inek sütünde bol miktarda miRNA bulunduğu tespit edilmiş ve inek sütü miRNA’larının gıda kalitesinde biyobelirteç olarak kullanım potansiyellerine yönelik bulgular elde edilmiştir. Ayrıca, güncel araştırmalar beslenme yoluyla inek sütü miRNA’larının insana transfer olarak önemli bir biyoaktif besin komponenti olabileceğini göstermektedir. Süt ve süt ürünlerinde üretim aşamalarında bozunmadan kalan inek sütü miRNA'larının insanların dolaşım sistemine geçerek farklı insan hastalıkları ile ilişkili önemli yolaklara etki edebileceği düşünülmektedir. Bu sebepten süt ve süt ürünlerinin miRNA içeriklerinin belirlenmesi önemlidir ve bu konuda güncel literatürde önemli bir boşluk olduğu gözlenmektedir. Bu çalışmada, literatürden insan homolog sekansına sahip ve inek sütünde bol miktarda bulunan miR-191’in kaynatma aşaması sonrası içme sütündeki miktarındaki değişiklik araştırılmıştır. Bu kapsamda süt örnekleri (çiğ süt ve pastörize süt) 100°C’de kaynatılarak örneklerden total RNA izolasyonu gerçekleştirilmiş ve elde edilen RNA’lardaki miR-191 miktarı RT-qPCR yöntemi ile analiz edilmiştir. Literatürde içme sütünün üretiminde kullanılan homojenizasyon ve pastörizasyon işlemlerinin miRNA spesifik farklı etkilere sebep olduğu gözlenmiştir. Gerçekleştirilen işlemler sonucunda literatüre uyumlu biçimde miR-191 miktarında kaynatılmış çiğ sütte %95.8 oranında (p<0.0001) ve kaynatılmış pastörize sütte %66.4 oranında (p=0.001) azalma gözlemlenmiştir. Bunun yanında çiğ süt ve pastörize sütte analiz edilen miR-191 için elde edilen CT değerleri arasında istatistiksel olarak anlamlı bir farklılık olduğu gözlenmiştir (p<0.0001). Çalışmamızın sonucu, sütün kaynatılmasının süt miRNA içeriği üzerindeki etkilerine ilişkin ön veriler ortaya koyarak işleme adımlarının süt miRNA bileşimi üzerine miRNA spesifik etkisinin olduğunu önemle vurgulamaktadır.

Keywords

miRNA, çiğ süt, pastörize süt, kaynamış süt, RT-qPCR, miR-191

Ethical Statement

Bu çalışmada etik kurul onayına ihtiyaç yoktur.

Supporting Institution

Bu çalışma Bursa Uludağ Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon birimi tarafından desteklenmiştir (FLO-2023-1551)

Project Number

FLO-2023-1551

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