人乳汁中microRNA的稳定性研究

为了研究人乳汁中的microRNAs(miRNAs)在体外储存状态下的稳定性。本研究采用模拟体外乳汁储存条件,对乳汁进行室温孵育24h,反复冻融6次,100℃孵育10min,以及模拟体内消化环境(RNA酶A/T137℃孵育1h)的处理条件,通过实时荧光定量PCR(qRT-PCR)方法检测处理前后乳汁中9个内源性免疫相关和1个外源性人工合成miRNA的相对表达量变化差异。研究结果显示:处理前后乳汁中内源性miRNAs和外源性miRNA均显著性降解(p<0.01)。内源性miRNAs降解为初始量的25%～70%之间,而外源性miRNA则急剧降解至初始量的0.06%～0.96%。我们的实验表明人乳汁中内源性的miRNAs较外源性miRNAs在体外不同环境下具有更强的稳定性。

Studies on the Stability of miRNAs in Human Breast Milk

In order to investigate the stability of microRNAs(miRNAs) in human breast milk,the raw breast milk was subjected to four types of treatment:incubated at 26℃ for 0.5 to 24 hours;subjected to six freeze-thaw cycles at-20℃;treated with RNase A and RNase T1 for 60 min at 37℃;incubated at 100℃ for 10 min.We profiled the expression of 9 endogenous miRNAs and 1 exogenous synthetic miRNAs between different treatments with control using real-time PCR(qRT-PCR).The result showed that the endogenous miRNAs and exogenous synthetic miRNAs were rapidly degraded(p<0.01).Compared to control,the endogenous miRNAs degraded to 25%～70%,whereas the exogenous synthetic miRNAs degraded to 0.06%～0.96%.These results indicated that the endogenous miRNAs should have resistance to the harsh conditions to a certain extent.