Let-7f-5p and application of target gene thereof for preparing medicine for treating osteoporosis
A technology of let-7f-5p, 1.let-7f-5p, applied in the application field of drugs for the treatment of osteoporosis, to achieve the effect of reducing bone loss
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Embodiment 1
[0041] High-throughput sequencing screening.
[0042] 1. High-throughput sequencing
[0043] 1.1 Method.
[0044] Using high-throughput sequencing, we screened the differential miRNAs of vertebral body tissues in the GIOP patient group and the control group (non-GIOP patients) from the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, and predicted and verified their target genes. At the same time, predict differential miRNAs target genes in MiRanda, MiRBase and TargetScan databases.
[0045] 1.2 Results.
[0046] The result is figure 1 As shown, figure 1 It is the cluster analysis diagram of miRNA between the GIOP patient group and the control group, where: GIOP is the GIOP patient group, and CON is the control group. It was found that miRNA let-7f-5p was differentially expressed.
[0047] 2. qRT-PCR verification
[0048] 1.1 Method.
[0049] The qRT-PCR method was used to verify and compare the expression of miRNA let-7f-5p between the GIOP patient grou...
Embodiment 2
[0054] The animal experiment verification of miRNA let-7f-5p as a medicine for treating osteoporosis.
[0055] 1. Let-7f-5p promotes osteogenic differentiation.
[0056] 1 method.
[0057] The long bones (tibia and femur) of 8-week-old mice were aseptically removed, and the bone marrow was washed with α-MEM containing 20% fetal bovine serum. Filtered with a 40μm cell strainer to obtain mouse bone marrow derived-mesenchymal stem cells (BMSCs), which were placed in a 35mm culture dish at 4×10 4 / cm 2 The density is at 37℃, 5% CO 2 Cultivate for 4 days.
[0058] The above-mentioned cultured BMSCs cells were seeded into a 12-well plate, and osteogenic induction medium (in a-MEM containing 25mg / mL L-ascorbic acid, 5mM β-glycerophosphate, 10% FBS) was used to induce osteogenic differentiation, and let -7f-5p mimics (single-stranded chemically enhanced oligonucleotides, let-7f-5p mimics, purchased from Guangzhou Ruibo Biotechnology Co., Ltd.) intervention, that is, let-7f-5p mimics (to ac...
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