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Application of inhibitor of lmo4 gene expression in preparation of psoriasis external application type therapeutic drug

A technology for gene expression and therapeutic drugs, which is applied in the field of preparation of topical psoriasis therapeutic drugs for external use, can solve the problems of high drug prices, shortages, and unsatisfactory therapeutic effects, and achieve convenient administration, high safety, and applicable strong effect

Active Publication Date: 2021-10-08
ANHUI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the targets of these biological agents are mainly concentrated in the immune link of psoriasis pathogenesis, in the process of medication, it may cause the intervention of the patient's immune system, and there are more adverse reactions for patients with moderate and severe psoriasis It is difficult to tolerate, and the price of these drugs is relatively high, which limits the use of these drugs
[0005] Therefore, the existing psoriasis treatment mainly focuses on the first two links, and the therapeutic effect is not satisfactory, lacking the last (third) link (target) for its pathogenesis, that is, the lack of excessive proliferation and abnormality of keratinocytes. Methods and drugs for differentiating interventions and treatments

Method used

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  • Application of inhibitor of lmo4 gene expression in preparation of psoriasis external application type therapeutic drug
  • Application of inhibitor of lmo4 gene expression in preparation of psoriasis external application type therapeutic drug
  • Application of inhibitor of lmo4 gene expression in preparation of psoriasis external application type therapeutic drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1. Immunohistochemical detection of lmo4 gene expression product LMO4 protein in skin lesions of psoriasis patients

[0051] 34 cases of clinically and pathologically confirmed psoriasis skin lesions were collected as experimental samples, all of which were in the ongoing or stable stage, including 15 females, 19 males, and 5 cases with family history; normal skin samples from clinical circumcision were collected 4 cases of tissue were normal samples. The expression of lmo4 gene expression product LMO4 protein in normal skin tissue and skin lesion tissue of patients with psoriasis was detected by immunohistochemical method.

[0052] The specific method includes the following steps:

[0053]1) Making tissue wax blocks: soak the experimental samples and normal samples in 4% (V / V) formaldehyde solution immediately, rinse the tissues with running water for 10 hours after 24 hours, then put them into a dehydrator for dehydration, and then put them in Soak in xylene...

Embodiment 2

[0062] Example 2. Isolation, culture and in vitro differentiation of normal human skin keratinocytes and detection of differentiated cell marker molecules and lmo4 gene expression

[0063] Example 1 The lmo4 gene expression product LMO4 protein is highly expressed at the tissue level of psoriasis by immunohistochemical methods, and the expression level change of the lmo4 gene in the differentiation process of normal skin keratinocytes is further detected by the following method Condition.

[0064] 1. Primary culture of keratinocytes

[0065] Basal layer keratinocytes were isolated from normal human skin tissue samples for primary culture.

[0066] The primary culture method comprises the following steps:

[0067] 1) Preparation before the experiment: preparation method of rat tail collagen and pretreatment of petri dishes: take 4 rat tails, wash the epidermis with detergent, then soak in 75% (V / V) alcohol for 30 minutes, and put them on the ultra-clean workbench Inside, rin...

Embodiment 3

[0088] Embodiment 3, human skin keratinocyte cell line HaCaT cell culture and differentiation in vitro

[0089] In order to further explore the effect of abnormal expression of lmo4 gene in keratinocytes on their proliferation and differentiation, the human skin-derived keratinocyte line HaCaT cells were selected for follow-up experiments.

[0090] The human skin-derived keratinocyte cell line HaCaT cell culture and in vitro differentiation method comprises the following steps:

[0091] 1) Resuscitate the human skin keratinocyte cell line HaCaT (purchased from Life Technologies, USA) in a water bath at 42°C. After the cells are completely adhered to the wall, replace the cells with K-SFM medium without calcium ions;

[0092] 2) Induction of differentiation: After the cells become undifferentiated long spindle-shaped, induce the cells at 37°C with DMEM medium containing 2 mmol / L calcium ions and 10% calf serum (FBS, purchased from Gibco) Differentiation, the time point of indu...

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Abstract

Provided is the application of human LIM domain only 4 gene (lmo4) in the preparation of psoriasis (psoriasis) external application type treatment medicine. The active ingredient of the drug is a recombinant lentiviral vector inhibiting the expression of the lmo4 gene or a recombinant lentivirus inhibiting the expression of the lmo4 gene.

Description

technical field [0001] The present invention belongs to the new use of lmo4 gene in the field of biotechnology, and specifically relates to inhibiting the expression of lmo4 gene to intervene and inhibit the excessive proliferation and abnormal differentiation of skin keratinocytes of patients with psoriasis, which can be used to prepare external-use local treatment for psoriasis disease (psoriasis) treatment drugs. Background technique [0002] Psoriasis (commonly known as "psoriasis"), with erythema, scales as the main manifestations, itching, easy to infection, easy to relapse. Psoriasis is a common chronic inflammatory disease of the skin, which is highly hereditary and accompanied by autoimmune features, and is more common in young adults. At present, the incidence of psoriasis in my country has reached 0.12%, and the number of patients is close to 10 million; in terms of worldwide statistics, the incidence of psoriasis accounts for 0.1% to 3% of the world's total popu...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12N15/113C12N15/86A61P17/06
CPCA61K48/00C12N15/113C12N15/86
Inventor 周海胜张学军
Owner ANHUI MEDICAL UNIV
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