CN116942702A - Application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury used in uterine transplantation - Google Patents
Application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury used in uterine transplantation Download PDFInfo
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- CN116942702A CN116942702A CN202311059575.6A CN202311059575A CN116942702A CN 116942702 A CN116942702 A CN 116942702A CN 202311059575 A CN202311059575 A CN 202311059575A CN 116942702 A CN116942702 A CN 116942702A
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- reperfusion injury
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- ischemia reperfusion
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- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims abstract description 23
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses an application of an exosome secreted by human umbilical cord mesenchymal stem cells as an anti-ischemia reperfusion injury used in uterine transplantation, belongs to the technical field of cell biology, and is an anti-ischemia reperfusion injury preparation used in uterine transplantation independently.
Description
Technical Field
The invention belongs to the technical field of cell biology, and particularly relates to application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury used in uterine transplantation.
Background
Uterine transplantation is the only treatment available for persons suffering from absolute infertility. However, due to the abundant and complex blood vessels around the uterus, the time consumption of the uterus transplantation operation is longer than that of other organ transplantation operations, and the organs face longer ischemia time and more serious ischemia reperfusion injury after operation. This exacerbates the risk of graft necrosis and loss of function following uterine implantation.
Ischemia reperfusion injury occurs mainly after blood reperfusion, resulting from the release of reactive oxygen species during reperfusion. The effects of ischemia reperfusion injury on the uterus are manifested in a number of ways, mainly by causing tissue peroxidation, causing aseptic inflammation, causing cell and tissue necrosis, causing microvascular dysfunction, and the like.
The main methods for dealing with ischemia reperfusion injury in organ transplantation at present are improving organ preservation solution and anti-ischemia reperfusion injury drug treatment. Both of these approaches inevitably employ antioxidants and certain immunosuppressants. However, most antioxidants and immunosuppressants have strong toxic and side effects, so that the clinical application of the antioxidants and immunosuppressants is limited. Finding anti-ischemia reperfusion injury drugs or biological agents with low toxic and side effects has been a research hotspot in the field of organ transplantation.
Exosomes refer to small vesicles (30-150 nm) containing complex RNAs and proteins, which are thought to be important mediators involved in intercellular communication, transporting cellular secretions. The exosomes secreted by the human umbilical cord mesenchymal stem cells are proved to have the effects of promoting tissue repair and improving the tissue oxidation resistance, and simultaneously have good tissue compatibility and are not easy to cause inflammatory reaction. However, the application and research of the exosome in the anti-ischemia reperfusion injury of uterine transplantation are yet to be developed.
Disclosure of Invention
The invention aims to solve the problems of complex operation process, longer ischemia time of uterine graft and higher ischemia reperfusion injury degree after uterine transplantation, and provides application of exosomes secreted by human umbilical cord mesenchymal stem cells with weaker immunogenicity as an anti-ischemia reperfusion injury preparation used after uterine transplantation.
The invention is realized by the following technical scheme:
the application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury agents used in uterine transplantation, the exosomes alone being anti-ischemia reperfusion injury agents used in uterine transplantation.
Further, the exosomes are used at a dose of 50-800 μg/mouse.
Further, the exosomes are used at a dose of 400 μg/mouse.
Compared with the prior art, the invention has the following advantages:
1. in a mouse uterine transplantation model, the ischemia reperfusion injury after a graft operation can be obviously relieved by using an exosome alone, the toxic and side effects are greatly reduced, and the method can make a great contribution to the survival and functional reconstruction of the graft after the operation of a patient and the improvement of the success rate of later pregnancy.
2. The invention is the application of the anti-ischemia reperfusion injury preparation used after the uterus transplantation of exosomes secreted by human umbilical cord mesenchymal stem cells for the first time.
Drawings
FIG. 1 is an image of a human umbilical cord-derived mesenchymal stem cell under the microscope of example 1 of the present invention;
FIG. 2 is a schematic diagram showing the lipid differentiation test of human umbilical cord-derived mesenchymal stem cells in example 1 of the present invention;
FIG. 3 is a schematic diagram showing the detection of osteoblast differentiation of human umbilical cord-derived mesenchymal stem cells in example 1 of the present invention;
FIG. 4 is an image of an exosome of human umbilical cord-derived mesenchymal stem cells secreted by an electron microscope in example 1 of the present invention;
FIG. 5 is a graph showing the particle size distribution of exosomes secreted by human umbilical cord-derived mesenchymal stem cells in example 1 of the present invention;
FIG. 6 is a photograph showing the steps of uterine implantation in example 2 of the present invention;
FIG. 7 is a graph showing the detection of the degree of oxidative stress in the grafts in example 2 of the present invention;
FIG. 8 is an inflammatory factor index detection in the graft in example 2 of the present invention;
FIG. 9 is a map of Masson staining of graft tissue in example 2 of the present invention.
Detailed Description
For a further explanation of the invention, reference is made to the following specific examples and figures.
Example 1
Observing the human umbilical cord mesenchymal stem cells under a microscope to observe the morphology, inducing the mesenchymal stem cells to differentiate into lipid and bone, judging the differentiation capacity, and extracting exosomes by using a gradient centrifugation method. And the morphology of exosomes was observed with a transmission electron microscope. The particle size distribution of the obtained exosomes was detected using a nanoflow.
As a result, the human umbilical cord mesenchymal stem cells were fusiform as shown in FIG. 1.
As shown in fig. 2 and 3, human umbilical cord mesenchymal stem cells undergo adipogenic and osteogenic differentiation.
As shown in fig. 4, the mesenchymal stem cell exosomes are microvesicles, and are round or oval in shape and regular in shape, as observed by a transmission electron microscope.
As shown in FIG. 5, the particle size of the mesenchymal stem cell exosomes of the present patent is mainly distributed between 50-120nm through nano-flow statistics.
Example 2
C57BL/6J female mice are selected as a transplanting donor, and a sleeve method is adopted for uterine transplantation molding. The schematic flow of modeling is shown in the technical scheme 6.
Before the graft is anastomosed with the receptor, the graft is placed in 4 ℃ normal saline for constant temperature preservation to simulate cold ischemia extension. The negative control group (NC) was not subjected to cold ischemia for a prolonged period, and was subjected to physiological saline injection through the tail vein from the 0 th day after the transplantation, with an injection dose of 200. Mu.L/mouse.d. The cold ischemia time of the positive control group (PC) was prolonged to 24 hours, and physiological saline injection was performed through the tail vein from the 0 th day after the transplantation, and the injection dose was 200. Mu.L/mouse.d. The cold ischemia time of exosome treatment group (EXO) is prolonged to 24h, the human umbilical mesenchymal stem cells are injected through tail vein from the 0 th day after the transplantation, the injection dosage is 400 mug/mouse.d, the injection volume is 200 mug/mouse.d, and physiological saline is selected as the exosome solvent.
The recipient mouse grafts and peripheral blood serum of each group were extracted 72h post-surgery. Part of the grafts were prepared into homogenates and assayed for SOD, CAT, MDA activity and content in the grafts, and the other part was tissue-sectioned for Masson staining. Peripheral blood serum IL-1. Alpha. And TNF-alpha. Concentrations in peripheral blood were measured by Elisa.
The receptor is weight (22+ -2) g, and female C57BL/6J (B6, H-2K) of 8-12 weeks old b ) Purchased from beijing vernalia laboratory animal limited. All experimental animals were bred in the SPF grade environment of Xiamen university organ transplantation institute and were anesthetized with pentobarbital sodium intraperitoneal injection.
Through the above experiments, as shown in fig. 7, the activities of antioxidant enzymes SOD and CAT in the implant treated by exosomes in this example were higher than those in the positive control group, and the content of lipid peroxide MDA was lower than that in the positive control group. This demonstrates that exosome treatment effectively reduces the degree of peroxidation of uterine grafts within 72 hours after blood supply recovery.
As shown in FIG. 8, the concentrations of the inflammatory factors IL-1. Alpha. And TNF-alpha in the peripheral blood serum of the recipient mice treated with the exosomes in this example were lower than those in the positive control group. This demonstrates that the treatment of exosomes effectively reduces the inflammatory response caused by the ischemic reperfusion injury of the uterine graft within 72 hours after the recovery of blood supply.
As shown in fig. 9, the degree of fibrosis of the uterine graft after the exosome treatment in this example is significantly lower than that of the positive control group, which indicates that the exosome treatment restores the uterine tissue to avoid tissue fibrosis caused by ischemia reperfusion injury, and has the potential of preventing uterine adhesion.
The foregoing description is only illustrative of the preferred embodiments of the present invention and is not to be construed as limiting the scope of the invention, i.e., the invention is not to be limited to the details of the invention.
Claims (3)
1. An application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury agents used in uterine transplantation, characterized in that exosomes are used alone as anti-ischemia reperfusion injury agents used in uterine transplantation.
2. Use of an exosome secreted by human umbilical cord mesenchymal stem cells as claimed in claim 1 as anti-ischemia reperfusion injury for use in uterine transplantation, wherein the exosome is used at a dose of 50-800 μg/mouse.
3. Use of an exosome secreted by human umbilical cord mesenchymal stem cells as claimed in claim 1 as an anti-ischemia reperfusion injury for use in uterine transplantation, wherein the exosome is used at a dose of 400 μg/mouse.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311059575.6A CN116942702A (en) | 2023-08-22 | 2023-08-22 | Application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury used in uterine transplantation |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311059575.6A CN116942702A (en) | 2023-08-22 | 2023-08-22 | Application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury used in uterine transplantation |
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| CN116942702A true CN116942702A (en) | 2023-10-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202311059575.6A Pending CN116942702A (en) | 2023-08-22 | 2023-08-22 | Application of exosomes secreted by human umbilical cord mesenchymal stem cells as anti-ischemia reperfusion injury used in uterine transplantation |
Country Status (1)
| Country | Link |
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| CN (1) | CN116942702A (en) |
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2023
- 2023-08-22 CN CN202311059575.6A patent/CN116942702A/en active Pending
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