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Method for establishing human endometritis model in vitro by using berberine to alleviate lipopolysaccharide

A technology for endometritis and endometrium, applied in the biological field, can solve the problem of no relevant reports on the efficacy of human endometritis

Pending Publication Date: 2019-07-26
THE FIRST HOSPITAL OF LANZHOU UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

At present, there is no relevant report on the curative effect of berberine in the treatment of human endometritis

Method used

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  • Method for establishing human endometritis model in vitro by using berberine to alleviate lipopolysaccharide
  • Method for establishing human endometritis model in vitro by using berberine to alleviate lipopolysaccharide
  • Method for establishing human endometritis model in vitro by using berberine to alleviate lipopolysaccharide

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Embodiment Construction

[0153] A method for establishing an in vitro human endometritis model by reducing lipopolysaccharide with berberine, comprising the following steps:

[0154] ⑴Isolation and culture of primary human endometrial stromal cells:

[0155] ①Take out the human endometrial tissue from the sample bottle filled with sterile normal saline, wash the human endometrial tissue 2~3 times with PBS, wash away the blood stains and secretions, and use ophthalmic scissors to cut the tissue to a diameter < 1mm;

[0156] ② Transfer the intima tissue obtained in step ① to a 15 mL centrifuge tube, add 5-7 mL type IV collagenase, digest in a 37°C water bath for 40-60 min, and shake the centrifuge tube every 10 min to speed up digestion , until the tissue becomes flocculent; then add 5-7 mL of DMEM / F-12 complete culture solution to stop the digestion;

[0157] ③ Filter once with a 100-mesh (150 μm) sieve to remove undigested tissue, then filter the filtered cell suspension A with a 400-mesh (40 μm) fi...

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Abstract

The invention relates to a method for establishing a human endometritis model in vitro by using berberine to alleviate lipopolysaccharide. The method comprises the steps of (1) isolation and culture of primary human endometrial stromal cells: using primary cell culture in vitro and the secondary screening method to extract 2-3 generations of human endometrial stromal cells; (2) utilizing the CCK8method to screen the appropriate concentration and time of LPS on the cells; (3) adopting the CCK8 method to screen the appropriate concentration and time of BBR on the cells; (4) detecting the content of IL-1 beta and TNF-alpha by ELISA; (5) data analysis. The CCK8 method is adopted to detect the toxic effects of different concentration gradients of LPS and BBR on hESCs at the different time, andthe ELISA method is utilized to identify an LPS-induced inflammation model to determine that berberine has a certain curative effect on human endometritis and is a potential drug for treating the endometritis.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a method for establishing an in vitro human endometritis model by using berberine to reduce lipopolysaccharide. Background technique [0002] Chronic endometritis (CE) is a persistent inflammatory lesion of the endometrium caused by bacterial invasion of the uterine cavity, and is characterized by abnormal plasma cell infiltration in the endometrial stroma. Studies have shown that the main pathogenic bacteria of CE are common bacteria, such as Escherichia coli, streptococcus, etc., followed by mycoplasma. [0003] The endometrium consists of two layers: the functional layer adjacent to the uterine cavity and the basal layer adjacent to the myometrium. Leukocytes can be found in the basal and functional layers, in luminal or intraepithelial locations of the gland. In the normal proliferative and early secretory phases of the menstrual cycle, leukocytes account for 10% of the...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2509/00C12N2501/90C12N2501/999
Inventor 马晓玲王一青骆晓荣
Owner THE FIRST HOSPITAL OF LANZHOU UNIV
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