Application of rapamycin in preparation of miscarriage prevention medicine for spontaneous abortion

A technology of rapamycin and spontaneous abortion, applied in the field of medicine

Pending Publication Date: 2020-06-19
THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no report about the application of rapamycin of the present inven

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  • Application of rapamycin in preparation of miscarriage prevention medicine for spontaneous abortion
  • Application of rapamycin in preparation of miscarriage prevention medicine for spontaneous abortion
  • Application of rapamycin in preparation of miscarriage prevention medicine for spontaneous abortion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Analysis of decidual macrophage resident ratio and adhesion molecule levels in normal early pregnancy and spontaneous abortion women

[0065] 1 Experimental method

[0066] The decidual tissues from normal early pregnancy and spontaneous abortion women were cut up and digested, and the decidual immune cells were isolated and analyzed by flow cytometry (all flow cytometry antibodies were from BioLegend).

[0067] 2 Experimental results

[0068] Compared with normal early pregnancy group, CD45 of spontaneous abortion women + , CD14 + The number of decidual macrophages decreased in the tissue (eg figure 1 As shown in A, P=0.0173), and accounted for CD45 + Decreased proportion of decidua immune cells (eg figure 1 As shown in B, P=0.0048). CD14 was then detected by flow cytometry + Proportion of adhesion molecules on decidual macrophages, including E-cadherin, VE-cadherin, vascular cell adhesion molecule 1, and L-selectin. The expression of the above adhesi...

Embodiment 2

[0069] Example 2 Analysis of autophagy levels of decidual macrophages in normal early pregnancy and spontaneous abortion women

[0070] 1 Experimental method

[0071] The decidual tissues from normal early pregnancy and spontaneous abortion women were taken, cut up, digested and separated to obtain decidual immune cells, and then sorted by magnetic beads (CD14 magnetic beads and analysis related reagents were purchased from Miltenyi Company) CD14 + Decidual macrophages. The average fluorescence density of the FITC channel was analyzed by flow cytometry using autophagy-lysosome green fluorescent dye (purchased from Shanghai Dongren Chemical Technology Co., Ltd.).

[0072] 2 Experimental results

[0073] It was found that the autophagolysosome fluorescence density of spontaneously aborted decidual macrophages was lower than that of the control group (such as figure 2 As shown in A, P=0.0286). After the enriched decidual macrophages were obtained, RNA was extracted and analy...

Embodiment 3

[0076] Example 3 In vitro experiments verify that rapamycin promotes the adhesion of macrophages to decidua stromal cells

[0077] 1 Experimental method

[0078] Primary decidual stromal cells were isolated from normal early pregnancy decidual tissue, purified and cultured, treated with live cell red dye (purchased from sigma company), seeded in a well plate at an appropriate density, and cultured overnight until they adhered to the wall. Human peripheral blood mononuclear cell line U937 cells were treated with phorbol ester (purchased from sigma company, 100ng / mL) for 24 hours to induce macrophages, and empty-loaded lentivirus with green fluorescence (purchased from Shanghai Jikai Gene) was used to induce macrophages. Macrophages were infected and treated with rapamycin (purchased from MCE Company) or a control solvent, and the in vitro adhesion experiment was performed.

[0079] 2 Experimental results

[0080] The results showed that rapamycin treatment increased the relat...

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Abstract

The invention relates to the technical field of medicines, in particular to application of rapamycin in preparation of a miscarriage prevention medicine for spontaneous abortion. Compared with normalpregnancy, macrophages residing in decidua tissue of a patient suffering from recurrent spontaneous abortion are reduced, and expression of adhesion molecules is low; and the autophagy level of the decidual macrophages of the abortion patient is low. Experimental results show that the rapamycin can significantly promote adhesion of the macrophages to decidua matrix cells. Normal pregnant rats andabortion pregnant rat models are constructed, and in-vivo experiments prove that: the rapamycin is applied to reduce dwelling of decidual macrophages caused by autophagy disorders, maintains the steady state of decidua immune components and significantly reduces the embryo absorption rate of the pregnant rats, thereby improving the outcome of bad pregnancy. Therefore, the rapamycin is expected tobe used as the miscarriage prevention medicine for spontaneous abortion, a new treatment scheme is provided for patients suffering from repeated spontaneous abortion, the psychological burden of the patients is relieved, and the rapamycin can be well applied clinically.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of rapamycin in the preparation of natural abortion medicine. [0002] Compared with normal pregnancy, the macrophages resident in the decidual tissue of patients with repeated spontaneous abortion were reduced, and the expression of adhesion molecules was lower; and the autophagy level of decidual macrophages in patients with abortion was low. Rapamycin can significantly promote the adhesion of macrophages to decidual stromal cells. Then, normal pregnant mice and aborted pregnant mouse models were constructed, and in vivo experiments confirmed that the application of rapamycin can maintain the homeostasis of decidual immune components by rescuing the decrease in decidual macrophage residence caused by autophagy disorders, and significantly reduce Embryo absorption rate in pregnant mice, thereby improving adverse pregnancy outcomes. Therefore, rapamycin is expect...

Claims

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Application Information

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IPC IPC(8): A61K31/436A61P15/06A23L33/10
CPCA61K31/436A61P15/06A23L33/10A23V2002/00A23V2200/30
Inventor 李明清杨慧丽李大金
Owner THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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