A kind of mirna-2861 antisense nucleotide pharmaceutical composition and its application
A technology of miRNA-2861 and antisense nucleotides, applied in the pharmaceutical composition of miRNA-2861 antisense nucleotides for the treatment of heart diseases, in the field of new drugs for endogenous non-coding small RNAs, and can solve the problem of unidentified key miRNAs, etc. problems, to achieve the effect of pharmacological reliability, reasonable formula and wide application range
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0017] The miRNA-2861 antisense nucleotide overexpression adenovirus vector and the negative control adenovirus were constructed. In this example, the miRNA-2861 antisense nucleotide sequence was artificially synthesized and subcloned into the pSilencer Adeno 1.0-CMV system of Ambion Company. Construction of miRNA-2861 antisense nucleotide overexpression adenovirus. Its miRNA-2861 antisense nucleotide sequence is shown in the following SEQ ID NO: 1: 5'-CCGCCCGCCGCCAGGCCCC-3'
[0018] According to the company's instructions, the Pac I linearized vector and the linearized adenovirus backbone (AdenovirTs LacZ Backbone) were co-transfected into HEK-293 cells, the adenovirus was packaged, the virus was amplified and collected, and the virus titer was determined; the empty vector Co-transfect HEK-293 cells with the adenovirus backbone according to the above steps, and package the negative control adenovirus.
[0019] Dilute the above obtained recombinant adenovirus phosphate buffer...
Embodiment 2
[0021] Changes in the expression level of miRNA-2861 under the stimulation of PE and Ang Ⅱ. For the hypertrophy model of cardiomyocytes in this example, the established method was used to cultivate the primary cardiomyocytes of rat suckling mice (rat suckling mice were purchased from Beijing Medical University, Da The mouse strain is Wistar, and the preparation of primary cardiomyocytes of rat suckling rats can be found in the following literature: W.-Q.Tan, et al, Foxo3aInhibits Cardiomyocyte Hypertrophy through Transactivating Catalase J BiolChem.2008October 31; 283(44):29730-29739) , the cardiomyocytes were treated with PE and Ang Ⅱ, and the total RNA of the cells was extracted at different times of culture, and the expression level of miRNA-2861 was detected by real-time fluorescent quantitative PCR technology, as shown in figure 1 As shown in A and 1B, among them, figure 1 A is the expression level of miRNA-2861 over time in primary cardiomyocytes of rat neonatal rats tre...
Embodiment 3
[0023] miRNA-2861 antisense nucleotides can inhibit hypertrophy induced by PE at the cellular level. In this example, primary cardiomyocytes were transfected with miRNA-2861 antisense nucleotides (anta-2861), which can effectively inhibit miRNA -2861 expression, at the same time the cells were treated with PE, and at the same time treated with the negative control (anta-NC) of miRNA-2861 antisense nucleotide, after 24 hours, the cardiac hypertrophy index was detected on the cardiomyocytes , such as the surface area of cardiomyocytes ( figure 2 A), protein / DNA ( figure 2 B) Detection, the experimental results prove that miRNA-2861 antisense nucleotides can effectively inhibit cardiomyocyte hypertrophy induced by PE.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More