CN105288599A - Application of heme oxygenase-2 in preparing preparations inhibiting organ transplantation immune rejection - Google Patents
Application of heme oxygenase-2 in preparing preparations inhibiting organ transplantation immune rejection Download PDFInfo
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- CN105288599A CN105288599A CN201510697954.7A CN201510697954A CN105288599A CN 105288599 A CN105288599 A CN 105288599A CN 201510697954 A CN201510697954 A CN 201510697954A CN 105288599 A CN105288599 A CN 105288599A
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- organ transplantation
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- heme oxygenase
- skin
- immune rejection
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Abstract
The invention relates to application of heme oxygenase-2 in preparing preparations inhibiting organ transplantation immune rejection, and discloses application of heme oxygenase-2 and genes thereof in preparing preparations inhibiting organ transplantation immune rejection. The preparation can be used for inhibiting organ transplantation immune rejection and can increase the organ transplantation survival rate, and an effective method is provided for treating chronic end-stage diseases of the liver, the kidneys, the heart and the like.
Description
Technical field
The present invention relates to the application of Heme oxygenase-2 in preparation suppression organ transplantation immunity repulsion preparation.
Background technology
Organ transplantation is the main method of the chronic whole Terminal Disease such as treatment liver, kidney and heart, suppresses rejection to be one of key factor of transplant organ long-term surviving.Along with progress of research, the reaction of transplant organ indigenous protective draws attention gradually to stoping the effect of transplant rejection.Protective effect mediates mainly through protective gene, finds on the smooth muscle cell and vascular endothelial cell of transplanting survival organ, have the expression of protective gene by research, and the transplant organ having rejection seldom or is not expressed.Heme oxygenase-2 (HemeOxygenase-2; HO-2) HO-2 is a kind of important endogenous anti-apoptosis factor; key protective effect is play to oxidative stress and cytokine mediated apoptosis; HO-2 plays an important role in the cerebral blood flow regulating and controlling neonatal pig and mice. the transgenic mice of this research by preparing HO-2; by finding the allograft of mouse skin, the survival rate of HO-2 skin of transgenic mice is significantly higher than wild-type mice skin survival rate.
Summary of the invention
The difficult problem that long-term surviving rate is low is caused in order to solve transplant organ because of immunological rejection, we have prepared the transgenic mice of HO-2, research HO-2 reduces the effect of immunological rejection in mouse skin allograft, and then provides HO-2 and the application of gene in preparation suppression organ transplantation immunity rejection preparation thereof.
The invention discloses the application of Heme oxygenase-2 in preparation suppression organ transplantation immunity rejection preparation.
Invention also provides the application of Heme oxygenase-2 gene in preparation suppression organ transplantation immunity rejection preparation.
More specifically, the application of carrier for expression of eukaryon in preparation suppression organ transplantation immunity rejection preparation of HO-2 is provided.
We construct the carrier for expression of eukaryon pEF1 α-IRES-AcGFP of HO-2, prepare HO-2 transgenic mice by embryo's microinjection technique.HO-2 skin of transgenic mice is transplanted on BALB/c mouse back, and wild type FVB mice is as negative control simultaneously.Observation HO-2 transgenic mice and the time-to-live of negative control skin, the quantitative comparison of immunohistochemical analysis inflammatory cell change.
Preparation of the present invention can be used for suppressing organ transplantation immunity rejection, improves the survival rate of organ transplantation, for the chronic whole Terminal Disease such as treatment liver, kidney and heart provide effective ways.
Accompanying drawing explanation
Fig. 1 is that HO-2 transgenic mice and wild-type mice skin graft survival rate contrast.
Fig. 2 is inflammatory reaction after HO-2 transgenic mice and wild-type mice skin transplantation.
Detailed description of the invention
The amplification of embodiment 1:HO-2 gene and the structure of carrier for expression of eukaryon pEF1 α-HO-2-AcGFP
Extract C57BL/6 Hippocampus of Mice total serum IgE, with TaKaRa Reverse Transcription box synthesis cDNA.Take cDNA as template.With F:5 '-GTGAATTCATGTCTTCAGAGGTGGAGACC (EcoRI)-3 ' (SEQIDNO.1), R:5 '-TAGGATCCCATGTAGTACCAGGCCAATAG (BamHI)-3 ' (SEQIDNO.2) is the 848bp fragment of upstream and downstream primer amplification HO-2, amplification condition: 94 DEG C of 4min, 94 DEG C of 40s, 60 DEG C of 40s, 72 DEG C of 40s, 72 DEG C of 10min, 30 circulations.Amplified production 1% agarose gel electrophoresis, to determine amplified production size, then the PCR primer of H0-2 is used EcoRI and BamHI double digestion with carrier pEF1 α-IRES-AcGFP (buying in Clontech company) after reclaiming purification simultaneously, 1% agarose gel electrophoresis is carried out after enzyme action, and the digestion products both reclaiming through gel purification test kit, by the mixed in molar ratio of the product of both enzyme action purification according to 3:1, 24h is connected with T4DNA ligase 37 DEG C, transform in DH5d competent cell, then the screening of positive colony is carried out, extract positive bacteria plasmid with plasmid extraction kit and carry out enzyme action qualification, Sangon Biotech (Shanghai) Co., Ltd. is sent to check order.
The foundation of embodiment 2:HO-2 transgenic animal model
The transgenic mice (Sai Ye biotech firm has assisted by Guangzhou) of HO-2 is prepared by microinjection technique.
Embodiment 3: mouse skin transplantation experiments
The HO-2 transgenic FVB mice that application is grown up and wild type FVB mouse skin are transplanted to adult BALB/c mouse back respectively, each time period respectively gets the survival rate that 10 positive Mus and wild-type mice observe skin, as shown in Figure 1, during 2d, HO-2 transgenic (Tg) cutify compares (A1 with wild type (W), A2), A1 skin survival rate comparatively A2 skin survival rate difference with insignificance; During 4d, B1 skin is B2 skin comparatively, and B1 skin survival rate is 70%, B2 skin survival rate is 40%, and both reach significant level at survival rate difference; 8d, C1 skin is C22 skin comparatively, and C1 skin survival rate is 50%, B2 skin survival rate is 20%, and both reach significant level at survival rate difference.And immunohistochemical experiment is carried out, the inflammatory reaction of both observations to the positive skin of 2d, 4d, 8dd and negative skin, and as shown in Figure 2,2d, transgenic (Tg) mice cutify is wild type (W) mice there was no significant difference comparatively; 4d, the inflammatory cell digital display work of transgenic mice cutify is less than wild type and transplants mouse skin inflammatory cell number; 8d, inflammatory cell number obviously increases, but the inflammatory cell digital display of transgenic mice cutify work is less than wild type transplanting mouse skin inflammatory cell number.Result shows, and the skin survival rate of HO-2 transgenic mice is significantly higher than matched group, and scytitis cell number is significantly lower than matched group.
Claims (3)
1. Heme oxygenase-2 suppresses the application in organ transplantation immunity rejection preparation in preparation.
2. Heme oxygenase-2 gene suppresses the application in organ transplantation immunity rejection preparation in preparation.
3. the carrier for expression of eukaryon of Heme oxygenase-2 suppresses the application in organ transplantation immunity rejection preparation in preparation.
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| CN201510697954.7A CN105288599A (en) | 2015-10-22 | 2015-10-22 | Application of heme oxygenase-2 in preparing preparations inhibiting organ transplantation immune rejection |
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| CN201510697954.7A CN105288599A (en) | 2015-10-22 | 2015-10-22 | Application of heme oxygenase-2 in preparing preparations inhibiting organ transplantation immune rejection |
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| CN105288599A true CN105288599A (en) | 2016-02-03 |
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| CN201510697954.7A Pending CN105288599A (en) | 2015-10-22 | 2015-10-22 | Application of heme oxygenase-2 in preparing preparations inhibiting organ transplantation immune rejection |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105727379A (en) * | 2016-02-25 | 2016-07-06 | 顾宇春 | Haem oxygenase drug eluting stent |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000012118A2 (en) * | 1998-08-28 | 2000-03-09 | President And Fellows Of Harvard College | Inhibiting cardiomyocyte death |
| CN1507348A (en) * | 2001-03-30 | 2004-06-23 | ɣ����ҽҩ��˾ | Carbon monoxide generating compounds for treatment of vascular, inflammatory and immune disorders |
| CN1545548A (en) * | 2001-06-21 | 2004-11-10 | ��˼����˹�����Ͽ���˹ҽҩ���� | Carbon monoxide improves outcomes in tissue and organ transplants and suppresses apoptosis |
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2015
- 2015-10-22 CN CN201510697954.7A patent/CN105288599A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000012118A2 (en) * | 1998-08-28 | 2000-03-09 | President And Fellows Of Harvard College | Inhibiting cardiomyocyte death |
| CN1507348A (en) * | 2001-03-30 | 2004-06-23 | ɣ����ҽҩ��˾ | Carbon monoxide generating compounds for treatment of vascular, inflammatory and immune disorders |
| CN1545548A (en) * | 2001-06-21 | 2004-11-10 | ��˼����˹�����Ͽ���˹ҽҩ���� | Carbon monoxide improves outcomes in tissue and organ transplants and suppresses apoptosis |
Non-Patent Citations (2)
| Title |
|---|
| 张维维等: "HO-1通过调节T细胞发挥对移植器官的保护作用", 《医学综述》 * |
| 张腾业等: "血红素氧合酶-2基因真核表达载体的构建", 《家畜生态学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105727379A (en) * | 2016-02-25 | 2016-07-06 | 顾宇春 | Haem oxygenase drug eluting stent |
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Application publication date: 20160203 |