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Application of miR-7 expression inhibitor in preparing medicines used for treating systemic lupus erythematosus

A lupus erythematosus and mir-7 technology, applied in the field of biomedicine, can solve the problem of no research on the effect

Inactive Publication Date: 2014-07-02
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the role of miR-7 in the pathogenesis of autoimmune diseases, especially SLE, has not been studied

Method used

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  • Application of miR-7 expression inhibitor in preparing medicines used for treating systemic lupus erythematosus
  • Application of miR-7 expression inhibitor in preparing medicines used for treating systemic lupus erythematosus
  • Application of miR-7 expression inhibitor in preparing medicines used for treating systemic lupus erythematosus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The expression of miR-7 in the PBMC of embodiment 1 SLE patient and HC, T cell, B cell, monocyte

[0032] Total RNA was extracted by Trizol method, and after reverse transcription, real-time quantitative PCR was used to amplify and measure miRNA expression. The TaqMan probe method was used, and the reagents were primers and kits commercialized by Applied Biosystems (ABI) (reverse kit: name FG, Taqman(r) microRNA RT Kit 200rxns, product number 4366596; PCR kit: name TaqMan Gene Expression Master Mix, Cat. No. 4369016; Reverse Primer and PCR Primer: Name MicroRNA Assays 50RT+150PCR, Cat. No. 4427975).

[0033] Four cases of newly treated SLE patients and HC were collected, and the expression of miR-7 in PBMC was detected. It was found that the expression of miR-7 in SLE patients was significantly higher than that in HC, with a statistical difference (0.86±0.03vs0.10±0.08, p=0.0001 ), in order to further clarify the differences in the expression of miR-7 on each cell su...

Embodiment 2

[0034] Example 2 Dual-luciferase reporter gene system verification of the predicted target gene of miR-7

[0035] The target genes of miR-7 were predicted by Targetscan, PicTar and miRanda software. According to whether the biological functions of these target genes were related to the pathogenesis of autoimmune diseases, 8 target genes were selected as miR-7 for verification. They are PTEN, FOXO1, CD200R1, XIAP, RELA, CD72, BCL2L11, IL17RB, respectively.

[0036] Clone the 3'-UTR sequence of the predicted target gene of miR-7, and add restriction sites XbaI (5'-TCTAGA-3') and NotI (5'-GCGGCCGC-3') at both ends of the primer. The primer sequences are as follows:

[0037]

[0038] The pmiRGLO vector (purchased from Promega) and the amplified sequence were subjected to XbaI and NotI double enzyme digestion. After the target fragments are recovered, the respective amplified fragments are connected to the carrier to construct a dual-luciferase reporter gene carrier carrying t...

Embodiment 3

[0048] The detection of target gene PTENmRNA level in the T of embodiment 3 SLE patient and HC, B cell

[0049] The RNA of T and B cells of SLE patients and HC were extracted respectively, and after being reverse transcribed into cDNA, the mRNA expression level of target gene PTEN was measured by fluorescent quantitative PCR, and GAPDH was used as an internal reference gene.

[0050] Primers were designed as follows:

[0051]

[0052] The PCR reaction conditions are as follows: 95°C pre-denaturation for 1 min, 95°C for 5 s, 60°C for 30 s, 30 cycles, 95°C for 1 min, 55°C for 1 min, and then increasing the temperature by 0.5°C every 10 s for a total of 81 cycles to 95°C. Melting curve. The result is as image 3 shown. Comparing the expression of target gene PTEN mRNA in T and B cells of newly treated SLE patients and HC, it was found that the expression of PTEN mRNA in B cells of SLE patients was lower than that of HC, with a statistical difference (0.92±0.24vs 2.20±0.49, ...

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Abstract

The invention discloses an application of a miR-7 expression inhibitor in preparing medicines used for treating systemic lupus erythematosus (SLE). The invention also provides SLE treatment medicines containing the miR-7 expression inhibitor. When the expression of miR-7 is inhibited in B cells, the mRNA level of PTEN is raised, and B cell proliferation function is reduced. Therefore, the miR-7 expression inhibitor can influence the B cell proliferation function through the regulation on the PTEN gene, such that an SLE-treating effect is achieved. Also, miR-7 participates in SLE pathogenesis through inhibiting a target gene CD200R1 thereof. Therefore, the inhibition on miR-7 expression assists in treating SLE.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of miR-7 expression inhibitors in the preparation of medicines for treating systemic lupus erythematosus. Background technique [0002] Systemic lupus erythematosus (SLE) is the most classic autoimmune disease, covering almost the disorder of the entire immune system. The immune system is composed of immune organs, immune cells and immune molecules. The immune response includes innate immunity and adaptive immunity. In recent years, studies have found that micro RNA (miRNA, micro RNA) is involved in the disorder of the immune system. [0003] miRNAs are a class of non-coding RNAs with 20-22 oligonucleotides. Mature miRNAs recognize the 3'UTR region of specific target mRNAs through nucleic acid sequence complementarity, and inhibit the expression of target mRNAs at the post-transcriptional level, thereby inhibiting protein synthesis and achieving regulation ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K48/00A61P19/04A61P37/02
Inventor 张烜李扬沈濂吴湘妮
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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