Antisense microrna-25 and its application
A microRNA-25, antisense technology, applied to antisense microRNA-25 and its application field
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Embodiment 1
[0025] A kind of antisense microRNA-25, it is the antisense RNA of microRNA-25, and it is the antisense RNA of microRNA-25, and its base sequence is as shown in SEQ ID NO.1, and the sequence of microRNA-25 is as shown in SEQ ID NO .2 shown.
[0026] The antisense microRNA-25 can be obtained by artificial synthesis, which is a conventional technical means.
Embodiment 2
[0028] In this embodiment, cobalt chloride (CoCl 2 ) and Oxygen Glucose Deprivation (OGD) model (OxygenGlucoseDeprivation, OGD) two methods to simulate in vitro hypoxia / oxidative stress conditions, respectively, to study the effects of two forms of oxidative damage on the expression of microRNA-25 in primary RPE cells and human RPE cell lines.
[0029] Primary RPE cells were deprived of glucose and oxygen for 12, 16, 18, and 20 hours, and reoxygenated for 2 hours. The expression of miR-25 was detected by qRT-PCR. It was found that the expression of miR-25 increased with the prolongation of glucose and oxygen deprivation.
[0030] After the primary RPE cells were treated with different concentrations of cobalt chloride solution for 12 hours, the expression of miR-25 was detected by qRT-PCR, and it was found that the expression of miR-25 was up-regulated with the increase of the treatment concentration.
[0031] Primary RPE cells were treated with 200 μM cobalt chloride solution...
Embodiment 3
[0035] Taking mouse oxygen-induced retinopathy (OxygenInduced Retinopathy, OIR) as the model of hypoxia / oxidative stress in vivo, the expression of microRNA-25 in the neural retina and RPE / choroid complex was detected by qRT-PCR in the normoxia group and the hypoxia group The expression difference was further confirmed by in vivo experiments on the induction effect of hypoxia / oxidative stress on microRNA-25.
[0036] Construction method of OIR mouse model:
[0037] On the 7th day after birth, the mice in the model group were placed in the animal experiment cabin connected to the oxygen meter along with 2 nursing mothers, and oxygen (O 2 ), so that the oxygen concentration is stably controlled at 75% ± 2%. Daylighting. Regularly open the oxygen chamber every day to clean, change the dressing (keep it dry), change food and add water, and replace the nursing mothers in the normal environment with the mothers in the cabin. On the 12th day after the mice were born, the mice in t...
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