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Application of JQ1 in preparing organ transplantation immunological rejection inhibiting medicines

A technology of immune rejection and immune exclusion, applied in the field of medical bioengineering, which can solve problems such as suppressing immune responses

Active Publication Date: 2018-06-01
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

JQ1 may potentially suppress the immune response

Method used

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  • Application of JQ1 in preparing organ transplantation immunological rejection inhibiting medicines
  • Application of JQ1 in preparing organ transplantation immunological rejection inhibiting medicines
  • Application of JQ1 in preparing organ transplantation immunological rejection inhibiting medicines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: The influence of JQ1 on the survival time of the transplanted heart was detected by constructing a mouse neck heterotopic heart transplantation model.

[0014] 1. Construction of mouse heterotopic heart transplantation model

[0015] 1.1 Experimental animals and main materials: The donor is a male pure-line C57 mouse, weighing 20-25g, purchased from Shanghai SLEC; the recipient is a male pure-line BALB / c mouse, weighing 20-25g, purchased from Shanghai Lecco Company. The arterial cannula is 26.

[0016] 1.2 Recipient preparation: The operation was performed under clean and non-sterile conditions, and anesthetized by intraperitoneal injection of 50 mg / kg of 1% pentobarbital sodium. After success, the mice were fixed in a supine position, and an oblique incision was made from the bottom of the right mandibular angle to the top of the midpoint of the right clavicle, and the right submandibular gland and right sternocleidomastoid muscle were removed layer by lay...

Embodiment 2

[0025] Example 2: Observation of pathological changes of heart grafts by HE staining.

[0026] 1. Experimental method: 1) On the 7th day after the transplantation, each 3 recipient mice in the control group and the experimental group were killed to obtain transplanted heart tissue pieces, which were fixed with 10% neutral formalin, Routinely embedded in paraffin, sliced ​​at 4 μm with a Leica microtome. 2) Sections were dewaxed with xylene and washed with various levels of ethanol to water. The dewaxing steps are: xylene (I) 5 min→xylene (II) 5 min→100% ethanol for 2 min→95% ethanol for 1 min→80% ethanol for 1 min→75% ethanol for 1 min→distilled water washing for 2 min. 3) Harris hematoxylin semen was stained for 5 minutes, and washed with tap water for 1 minute. 4) Differentiate in hydrochloric acid ethanol solution for 30s. 5) Soak in tap water for 15 minutes or warm water (about 50°C) for 5 minutes. 6) Place the eosin aqueous solution for 2 minutes. 7) Routine dehydrat...

Embodiment 3

[0028] Example 3: Detection of the expression changes of related inflammatory factors in the heart of mouse recipients by means of fluorescent quantitative PCR.

[0029] 1. Obtaining Heart Graft Specimens

[0030] Seven days after the transplantation, six mice in each of the control group and the experimental group were sacrificed, and the heart grafts were taken out, marked and stored at -80°C.

[0031] 2. Total RNA extraction (pipe tips and centrifuge tubes are sterilized by moist heat, no RNase)

[0032] 1) Take a homogenizer, add 1ml of Trizol Reagent, and pre-cool on ice.

[0033] 2) Take 100 mg of tissue and add it to a homogenizer.

[0034] 3) Grind thoroughly until no tissue pieces are visible.

[0035] 4) Centrifuge at 13000 g for 10 min to take the supernatant.

[0036] 5) Add 250 μl of chloroform, invert the centrifuge tube for 15 seconds, mix well, and let stand for 3 minutes.

[0037] 6) Centrifuge at 13000 g for 8 min at 4°C.

[0038] 7) Transfer the supernat...

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Abstract

The invention provides an application of JQ1 in preparing organ transplantation immunological rejection inhibiting medicines, in particular an application in preparing medicines for preventing and treating cardiac transplantation immunological rejection. Based upon experiments, it is proved that the survival time of transplanted hearts of an allogeneic mice (C57BL / 6 to BALB / c) in a control group is 7-10 days, while the survival time of transplanted hearts of mice applied with the JQ1 can reach 25 days to the greatest extent, indicating that the JQ1 provided by the invention is beneficial for reducing an alloimmunity reaction and prolonging the survival of transplants. The JQ1, as a novel small-molecule inhibitor, can solve the problem that transplanted organs are low in survival rate for along time due to rejection, and the JQ1 can offer a basis for preparing organ transplantation immunological rejection inhibiting preparations; therefore, an effective approach is provided for treating such chronic end-stage diseases such as liver diseases, renal diseases, cardiac diseases and the like.

Description

technical field [0001] The invention relates to the field of medical bioengineering, in particular to the application of JQ1 in the preparation of medicines for immune rejection of organ transplantation, especially the application in prolonging the survival of transplanted hearts. Background technique [0002] Heart transplantation is an effective means of treating various end-stage heart diseases. However, post-transplant rejection and related complications caused by anti-rejection therapy limit the long-term survival of recipients. The incidence of acute rejection has decreased with the use of newer immunosuppressants, but it remains a major cause of death in recipients within 1 year of heart transplantation. Acute cellular rejection (acute cellular rejection, ACR) often occurs 3-6 months after surgery or occasionally at any time after surgery, and is mediated by T cells to infiltrate the graft with lymphocytes and macrophages, which not only causes short-term graft damag...

Claims

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Application Information

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IPC IPC(8): A61K31/551A61P37/06
CPCA61K31/551
Inventor 龚渭华孙启明陈俊涛刘宝清刘晨
Owner ZHEJIANG UNIV
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