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Uses of mir‑135 and its related drugs

1. The technology of mir-135, which is applied in the direction of drug combination, animal cells, biological testing, etc., can solve the problem that the function of miR-135 has not been studied, and achieve the effect of high efficiency

Inactive Publication Date: 2017-10-03
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite so many current studies on miR-135, most of the functions of miR-135 have not been investigated, especially in kidney diseases (such as podocyte disease)

Method used

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  • Uses of mir‑135 and its related drugs
  • Uses of mir‑135 and its related drugs
  • Uses of mir‑135 and its related drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Upregulation of miR-135 expression in podocyte injury model

[0058] 1. The expression of miR-135 is upregulated in the mouse model of podocyte injury

[0059] To study the role of miR-135 in podocyte injury, we first established a mouse podocyte injury model with doxorubicin, sacrificed the mice at different time points after injecting doxorubicin into the tail vein, dissected out the kidneys, and isolated the nephrons. ball. Then use real-time fluorescent quantitative PCR to detect the expression changes of miR-135, such as figure 1 As shown, the expression of miR-135 increased on the fourth day after doxorubicin injection and peaked on the eleventh day.

[0060] 2. The expression of miR-135 is upregulated in the in vitro model of podocyte injury

[0061] To further study the role of miR-135 in cells in vitro, we detected the expression changes of miR-135 in podocytes after treatment with different concentrations of doxorubicin for different periods of ti...

Embodiment 2

[0064] Example 2 Overexpression of miR-135 in the mouse podocyte line MPC5 can lead to podocyte damage

[0065] 1. Overexpression of miR-135 can lead to up-regulation of podocyte injury markers and down-regulation of functional markers

[0066] The cultured MPC5 cells were transfected with miR-135mimics, and after 48 hours, the cells were lysed to extract RNA and protein. As we all know, desmin and snail are two classic injury markers of podocytes, while nephrin, E-cadherin and WT1 are functional markers of podocytes. Quantitative PCR and western blot techniques were used to detect the expression changes of some related genes in podocytes. The detection results Such as Figure 4 (RNA level) and Figure 5 As shown (at the protein level), the expression of desmin and snail was upregulated, while the expression of nephrin, WT1 and E-cadherin was decreased.

[0067] 2. The overexpression of miR-135 can cause the disorder of podocyte cytoskeleton

[0068] Recently, it has been ...

Embodiment 3

[0069] Example 3 miR-135 can activate the wnt signaling pathway

[0070] 1. miR-135 can activate the activity of the Topflash reporter carrier

[0071] Figure 4 with Figure 5 It was shown that miR-135 can promote the expression of desmin and snail, and these two genes are the target genes downstream of the wnt signaling pathway, so we guess whether miR-135 can activate the wnt signaling pathway, and the activation of the wnt signaling pathway has been reported in the literature to lead to sufficient cell damage. Based on these studies, we first investigated the effect of miR-135 on Topflash activity in MPC5 and 293T cells, and the results were as follows Figure 7 As shown, miR-135 can activate Topflash activity in both MPC5 cells and 293T cells, and LiCl was used as a positive control.

[0072] 2. miR-135 can promote the expression of unphosphorylated active β-catenin

[0073] Total protein was extracted 48h after overexpressing miR-135 in MPC5 cells, and the expressio...

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Abstract

The invention relates to the field of biotechnology, in particular to a purpose and relevant medicine of miR-135. The invention discloses the dependency of the miR-135 and the foot cell injury for the first time, and discloses the capability of overexpression miR-135 to cause foot cell injury for the first time, and the process is more similar to the human disease causing process. A foot cell injury model has the sufficient foot cell injury typical features, can well simulate the occurrence and the development of foot cells in the body, and belongs to an ideal model of foot cell injury base and clinic application study, so that the miR-135 can be well applied to the medicine for screening and treating the foot cell injury.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of miR-135 and related medicines. Background technique [0002] As a class of small (18-25 nucleotides) non-coding RNA, microRNA plays a crucial role in regulating gene expression in the life activities of various organisms. In mammals, more than 1,000 microRNA sequences have been discovered, each microRNA can regulate multiple messenger RNAs (mRNA), and each mRNA can be regulated by hundreds or thousands of microRNAs. Studies estimate that over 60% of protein-coding gene expression is regulated by microRNAs. [0003] Recent studies have found that microRNA can abnormally regulate gene expression in kidney diseases. For example, miR-93 can promote glomerular injury by activating vascular endothelial growth factor; miR-195 can promote podocyte apoptosis by inhibiting the expression of BCL2; and accelerate podocyte injury by down-regulating miR-30. miR-192 can promote the fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N5/071C12Q1/02A61K48/00A61K31/7105A61P13/12G01N33/53
CPCA61K31/7105C12Q1/6883C12Q2600/178G01N33/5044
Inventor 周钦吕小岩李千音谢亚均吕中石
Owner CHONGQING MEDICAL UNIVERSITY
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