Application of microRNA-210 inhibitor in preparing medicine treating inflammatory dermatosis

A technology of microrna-210, 1.microrna-210, which is applied in the field of biomedicine and can solve problems such as uncertainty

Active Publication Date: 2018-06-12
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, whether these miRNAs molecules can be applied to the preparation

Method used

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  • Application of microRNA-210 inhibitor in preparing medicine treating inflammatory dermatosis
  • Application of microRNA-210 inhibitor in preparing medicine treating inflammatory dermatosis
  • Application of microRNA-210 inhibitor in preparing medicine treating inflammatory dermatosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1, the expression of microRNA-210 is increased in IMQ-induced inflammatory lesions of mice

[0042] Select 12 7-week-old, healthy female Balb / c mice with back hair removal 2×2cm 2 Afterwards, they were randomly divided into two groups: the first group was the model group, and each mouse was coated with 5% imiquimod ointment 62.5mg / d on the depilated area of ​​the back every day, for 6 consecutive days; the second group was the normal group, each mouse Apply matrix ointment 62.5 mg / d to the depilated area on the back of the mouse every day for 6 consecutive days. On the 7th day after modeling, the mice in each group were killed by neck breaking, and the skin lesions on their backs were taken, fixed in formalin solution overnight, dehydrated, embedded in paraffin, sliced ​​for HE staining, and histopathologically observed under a microscope Change. The results showed that compared with the normal group mice, the skin of the mice in the model group had visible sc...

Embodiment 2

[0045] Example 2. Research on the influence and mechanism of abnormal expression of microRNA-210 on keratinocytes and T cells

[0046] 1) microRNA-210 promotes keratinocyte proliferation

[0047] In this experiment, primary human keratinocytes were used. 12 hours (hours, hrs) before transfection, the primary human keratinocytes were seeded in 96-well plates, and cultured with corresponding 100 μL serum-free at 37°C, 5% CO 2 Cultivate in the incubator for 12h. When the cells grow to 90% confluence, transfect agomiR-210 (microRNA-210 mimic), antagomiR-210 and the corresponding negative control into keratinocytes, replace the fresh medium after 6hrs, and continue to culture for 24hrs or 48hrs . 4 hrs before the end of the culture, 10 μL of CCK8 solution was added to each well to continue the culture for 4 hrs. Cell proliferation was detected at a wavelength of 450 nm. The results showed that overexpression of microRNA-210 could significantly promote the proliferation of prim...

Embodiment 3

[0063] Example 3. Overexpression of microRNA-210 in vivo can significantly promote and aggravate IMQ-induced inflammatory skin lesions in mice

[0064] Select healthy female Balb / c mice aged 6-8 weeks, with hair removal on the back of 2×2cm, and randomly divide them into two groups: 1) agomiR-210 group: apply 5% IMQ ointment 62.5mg / d on the back every day, for 6 consecutive days , and intradermally injected agomiR-210 150 μl (5 nmol) on the back on days 0, 1, 2, and 3; 2) agomiR-NC group: 5% IMQ ointment 62.5 mg / d was applied externally on the back every day for 6 consecutive days, and On day 0, 1, 2, and 3, 150 μl (5 nmol) of agomiR-NC was injected intradermally on the back. Three mice in each group were sacrificed on the 4th, 7th, 10th, and 14th day of modeling, and the clinical and pathological changes of the skin lesions were observed, and histological analysis was performed. The expression of microRNA-210 was detected by real-time PCR. Expression changes. The results sh...

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Abstract

The invention discloses application of microRNA-210 inhibitor in preparing medicine treating inflammatory dermatosis. A large quantity of experiments prove that in-vitro inhibiting of expression of microRNA-210 can obviously improve the expression of a target gene STAT6 of microRNA-210, thus the formation of cell proliferation by cutin and secretion of chemokine CCL20 are inhibited, migration of chemotaxis T cells of chemokine CCL20 to the skin lesion parts is further inhibited, and meanwhile, differentiation of TH1 and TH17 can be inhibited. MicroRNA-210 knockout and injection of the microRNA-210 inhibitor (antagomiR-210 modified by cholesterol) given into the skin lesion specifically inhibits the expression of microRNA-210, which can both obviously inhibit mouse skin inflammation and improve T cell immune imbalance. A novel pathophysiology mechanism is provided for inflammatory dermatosis, and a new strategy capable of being used for preparing the medicine treating inflammatory dermatosis is provided.

Description

technical field [0001] The invention relates to a new medical application of an endogenous non-coding small RNAs inhibitor, specifically the application of a microRNA-210 inhibitor in the preparation of drugs for treating inflammatory skin diseases, which belongs to the field of biomedicine. Background technique [0002] Inflammatory skin diseases are a common type of skin diseases mediated by immune cells and keratinocytes. Abnormal response of innate immune system, abnormal activation of T lymphocytes, various inflammatory cytokines and their target cells (keratinocytes) play an important role in the pathogenesis of inflammatory skin diseases. Among them, a series of inflammatory reactions caused by helper T cell (Th cell) immune imbalance are relatively common in inflammatory skin diseases. Th cells are stimulated by some antigen-presenting cells and natural immune cells, and the immune balance among various cell subsets is disturbed, resulting in abnormal secretion of i...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/7088A61P17/00A61P17/06
CPCA61K45/00A61K31/7088A61K31/713A61P17/00A61P17/06C12N15/113C12N2310/113C12N2310/3515Y02A50/30C12N2310/141C12N2310/32C12N2320/34
Inventor 陆前进赵明苏玉文武瑞芳
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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