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Experimental method for treating POI by combining ASP with hAD-MSCs

An experimental method, the had-mscs technology, is applied in the field of ASP combined with hAD-MSCs in the treatment of POI, which can solve the problems of low migration homing rate and local survival rate and limit the treatment effect, and achieve the promotion of survival rate, good promotion prospect and improvement of proliferation Effect

Pending Publication Date: 2022-02-08
何连利
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Problems solved by technology

[0008] MSCs transplantation for the treatment of POI caused by RT has become a research hotspot, but its low migration homing rate and local survival rate limit its therapeutic effect.

Method used

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  • Experimental method for treating POI by combining ASP with hAD-MSCs

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Embodiment

[0039] An experimental method for treating POI with ASP combined with hAD-MSCs, comprising the following steps:

[0040] Specifically include the following steps:

[0041] 1) Preparation of ASP drug-containing serum: the dosage of Angelica sinensis containing polysaccharides was converted into the dosage of rats according to the clinical dosage, and the dosage of Angelica polysaccharides was converted into the relative dosage according to the same period of the previous year; 2 hours after the injection, the rats were killed, blood was collected from the heart, serum was collected by conventional methods, complement was inactivated at 56°C for 30 minutes, sterilized by 0.22μm filtration, and stored at -20°C for later use;

[0042] 2) Different concentrations of ASP drug-containing serum were added to the culture medium to cultivate hAD-MSCs, and the optimal concentration of ASP drug-containing serum was explored, and hAD-MSCs were identified and purified;

[0043] 3) hAD-MSCs...

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Abstract

The invention relates to an experimental method for treating POI by combining ASP with hAD-MSCs. The experimental method comprises the following steps: preparing ASP drug-containing serum; adding ASP drug-containing serum with different concentrations into a culture medium to culture hAD-MSCs; and carrying out hAD-MSCs transplantation; 5) carrying out experiment grouping; observing ultrastructure change of each level of follicles and endometrium by using an electron microscope; carrying out immunohistochemical determination on the expression of a granular cell surface marker Fshr and an oocyte surface marker DDX4; and using an Elisa method for determining secretion conditions of antioxidant enzymes and oxidation products ROS, SOD and the like of the ovary and uterus tissues; (13) carrying out Western-blot determining and obtaining the change condition of the phosphorylation level; determining apoptosis conditions of follicular granular cells and oocytes by Tunel staining; determining the expression of the ovarian tissue FOXO3a through immunofluorescence double labeling; and 16) carrying out a cage closing experiment. The method has the advantages that the angelica polysaccharide is combined with the hAD-MSCs to improve the proliferation of stem cells, the microenvironment of the radioactive POI can be improved, the migration homing and local survival rate of the stem cells can be promoted, the treatment effect of the stem cells on the radioactive POI can be improved, and the method has a good popularization prospect.

Description

technical field [0001] The invention relates to an experimental method for treating POI with ASP combined with hAD-MSCs. Background technique [0002] Ovarian insufficiency (POI) caused by radiotherapy (Radiotherapy RT). RT is a common treatment modality for many types of cancer, with approximately 70% of all cancer patients receiving RT as part of their disease treatment. Radiation can directly or indirectly cause damage to multiple organs in the body, among which germ cells with vigorous metabolism and rapid renewal are most likely to be damaged by radiation. Previous studies have shown that when the ovary is exposed to radiation, the radiation damages germ cells through the following effects: 1) oocytes will be lost through cell death pathways; 2) granulosa cell damage leads to impaired production of gonadal hormones; 3) ovarian blood vessels and interstitial cells quality is damaged. These injuries lead to POI and even premature ovarian failure (POF), which is the ter...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K31/715A61P15/00
CPCA61K35/28A61K31/715A61P15/00A61K2300/00
Inventor 何连利谢红梅张韵函莫晓莉张青云祝叶龙小琴
Owner 何连利
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