Application of reagent for down-regulating Czib gene expression in preparation of medicine for treating or improving pulmonary fibrosis
A gene expression and pulmonary fibrosis technology, applied in the field of biomedicine, can solve problems such as obvious side effects and achieve the effect of inhibiting the occurrence of pulmonary fibrosis
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Embodiment 1
[0059] Expression changes of Czib in a bleomycin-induced mouse model of lung fibrosis.
[0060] The construction of the bleomycin mouse model: the mice were anesthetized by intramuscular injection of pentobarbital sodium (30 mg / kg). Draw out 0.3ml of bleomycin (Bleo) normal saline solution (dissolved in 0.2-0.3 normal saline according to the ratio of 5 mg / kg) with the syringe, align the needle tip of the syringe with the cricoid cartilage space, penetrate the trachea and draw back the air, then raise the animal with the left hand With the head at a certain angle, push the needle with the right hand to inject slowly along the back wall of the trachea, and then continue to inject 0.3ml of air, then immediately erect the animal, rotate left and right, and pat the mouse on the chest and back to promote the uniform distribution of the drug in the lungs, the control group (Sal) Inject an equal volume of normal saline in the same way. After 21 days, the samples were taken, part of w...
Embodiment 2
[0062] Expression of Czib in bleomycin-induced A549 cells.
[0063] Construction of A549 cell injury model: A549 cells were treated with bleomycin for 72 hours, the total protein was extracted, and the expressions of Czib and fibrosis marker protein aSMA were detected by qPCR and Western blot. The result is as figure 2 The up-regulated expression of aSMA indicated that the induction of the A549 cell injury model was successful, and the expression of Czib was significantly up-regulated in this cell model.
Embodiment 3
[0065] Preparation of effective Czib siRNA.
[0066] Using siRNA design software, according to GenBank to obtain the Czib mRNA sequence of rats (NM_001304759.2), search the AA sequence and record the 19 nucleotides adjacent to the 3' end of each AA, and screen out the GC content between 30-55% siRNA. Further screening is carried out according to the siRNA design principles, and the homology of the screened siRNA sequences is searched by BLAST in the genome database of GenBank, and sequences with more than 3 base mismatches with non-homologous genes are selected to exclude For the possibility of non-specific inhibition, on the basis of meeting the above conditions, three siRNAs that inhibit the expression of Czib were screened out (see Table 1).
[0067] Table 1 siRNA sequences targeting Czib
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[0069]
[0070] Note: In the siRNA sequence, 5'-3' is the sense strand, and 3'-5' is the antisense strand.
[0071] Synthesize the siRNA sequences shown in Table 1, and...
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