CN-121999884-A - Method for researching relationship between intestinal tract fixation and oviduct edema after chlamydia genital tract infection and application

CN121999884ACN 121999884 ACN121999884 ACN 121999884ACN-121999884-A

Abstract

The invention discloses a method for researching the relation between intestinal tract field planting and oviduct edema after chlamydia genital tract infection and application thereof, belonging to the technical field of biological medicine. The invention provides a complete and standardized research system which can be used for researching the relation between intestinal tract fixation and salpingemphraxis after chlamydia genital tract infection, firstly reveals that the intestinal tract fixation and persistent and stable salpingoedema formation are necessary to be linked after chlamydia genital tract infection, provides a new direction for preventing and treating salpingoedema induced by chlamydia genital tract infection, shows that the method provided by the invention can be used for researching the relation between chlamydia intestinal tract fixation and salpingoedema and has reference value for other similar researches, and also provides a group of preparations which can be used for diagnosing and/or predicting risk and/or preventing and/or treating salpingoedema induced by chlamydia genital tract infection, wherein the preparations are marker flora obtained by identifying the method provided by the invention.

Inventors

ZHOU ZHOU
ZHOU YANG
TAN SHUI
CHEN LONG
HU YUCHEN
YANG WENJING
WEN XIN
TANG JIN

Assignees

南华大学

Dates

Publication Date: 20260508
Application Date: 20260122

Claims (10)

1. A method for researching the relationship between intestinal tract fixation and oviduct edema after chlamydia genital tract infection, which is characterized by comprising the following steps: S1, constructing a chlamydia genital tract infection animal model, dynamically monitoring chlamydia loading capacity of the animal model and analyzing the quantity and structure of intestinal flora; s2, constructing an animal model for the depletion of intestinal flora after chlamydia genital tract infection, and monitoring the fecal bacterial DNA content, the flora quantity, the weight change and the cecum condition of the animal model; S3, evaluating the influence of the depletion of intestinal flora on oviduct lesions induced by chlamydia genital tract infection; S4, constructing a chlamydia genital tract infection mouse model of fecal bacteria transplantation and remodeling, and detecting the number, weight and intestinal tract change condition of intestinal bacteria of the model animal; S5, evaluating whether intestinal flora can be rebuilt through fecal transplantation in a chlamydia genital tract infection animal model, and improving oviduct lesions; S6, screening marker species related to oviduct lesions induced by chlamydia genital tract infection.
2. The method for studying the relationship between intestinal tract colonization and tubal edema after chlamydia genital tract infection according to claim 1, wherein a pair of differential chlamydia strains and a pair of differential chlamydia tc0668 strains are adopted in the step S1 to respectively construct a genital tract infection animal model of chlamydia, wherein the chlamydia genital tract infection animal model is a mouse model, and the step of constructing the chlamydia genital tract infection animal model is as follows: following adaptive feeding, depo days prior to infection, mice were vaccinated with the resuscitated strain via the vaginal route in synchronization with the estrus cycle of the mice; The plasmid differential chlamydia strain is a chlamydia murine plasmid carrying strain Cm NiggII and a chlamydia murine plasmid deletion strain CM PLASMID-free, and the tc0668 differential chlamydia strain is a chlamydia murine tc0668 wild strain Cm G13.32.1 and a chlamydia murine tc0668 mutant strain Cm G28.51.1; Dynamic monitoring of chlamydia load in animal model by collecting vaginal and rectal swabs of mice on day 3, day 7, day 14 and following weekly, respectively, until day 56, and quantifying chlamydia load by indirect immunofluorescence, and taking mice inoculated with equivalent amount of SPG solution as control group.
3. The method for researching the relation between intestinal fixation and salpingemphraxis after chlamydia genital tract infection according to claim 1, wherein the analysis method for the intestinal flora structure in S1 is characterized in that feces of animals in different groups at different times after infection are collected, microbial DNA is extracted, PCR is carried out by adopting a primer specifically identifying a V4 region of a 16S rRNA gene, high-throughput sequencing is carried out on a PCR product, bioinformatics analysis is carried out on a sequencing result, the primer sequence specifically identifying the V4 region of the 16S rRNA gene is shown as SEQ ID NO.1 and SEQ ID NO.2, and the bioinformatics analysis comprises species classification unit division, alpha diversity evaluation, beta diversity evaluation, species composition analysis and inter-group biological marker identification.
4. The method for researching the relationship between intestinal tract colonization and tubal edema after chlamydia genital tract infection according to claim 2, wherein the method for constructing an animal model for intestinal flora depletion after chlamydia genital tract infection in S2 is characterized in that the mixture of vancomycin and gentamicin is infused on the 19 th day after the chlamydia differential strain genital tract infection and on the 3 rd day after the chlamydia differential strain genital tract infection in S1, the gastric lavage amount is 0.1mg/g of mice, and the mass ratio of the vancomycin to the gentamicin is 1:1.
5. The method for researching the relationship between intestinal colonization and salpingemphraxis after chlamydia genital tract infection according to claim 1, wherein the method for evaluating the influence of intestinal flora depletion on chlamydia genital tract infection induced oviduct lesions in S3 is to examine the oviduct disease transformation rate of chlamydia genital tract infection animals in a 60 th day of intestinal flora depletion group and chlamydia genital tract infection animals in S1, and the method for constructing a chlamydia genital tract infection mouse model by means of fecal fungus transplantation and remodeling in S4 is to collect fresh feces of chlamydia genital tract infection animals in S1 on 21 th day, prepare fecal fungus suspension and irrigate the chlamydia genital tract infection post-intestinal flora depletion animal model constructed in S2 every day.
6. The method for studying the relationship between intestinal tract colonization and salpingemphraxis after chlamydia genital tract infection according to claim 2, wherein the method for screening the differential marker species in S6 is characterized in that a LEfSe algorithm is applied to carry out deep analysis on LEfSe results in sequencing data of S1, and marker flora which is shared or not shared is screened between chlamydia virulent strain infection groups and between attenuated strain infection groups, wherein the chlamydia virulent strain infection groups are Cm NiggII infection groups and Cm G13.32.1 infection groups, and the attenuated strain infection groups are CM PLASMID-free infection groups and Cm G28.51.1 infection groups.
7. A preparation for preventing and/or intervening tubal edema, characterized in that the preparation comprises a bacterial population selected from the group consisting of a highly pathogenic bacterial population and a less pathogenic bacterial population according to any one of claims 1 to 6, wherein the highly pathogenic bacterial population is selected from the group consisting of helicobacter Helicobacteraceae, RF, burkholderiales Burkholderiales, wenyujin RIKENELLA, cyanobacteria Cyanobacteria, heterologous bacteria, 4C0d-2, erysipelas Erysipelotrichales, YS2, turicibacteraceae, verrucomicrobiales Verrucomicrobiales, verrucomicrobiales Verrucomicrobiae, beta proteobacteria, sarcandidia Sutterella, acremodella AKKERMANSIA, TURICIBACTERALES, erysipelas Erysipelotrichi, verrucomicrobiales Verrucomicrobia, verrucomicrobiaceae Verrucomicrobiaceae, cholangium RIKENELLACEAE, RF, corynebacteriaceae, rhodococcus Ruminococcus, rhodococcus rhodochrous 6569, rhodococcus 3695, and Staphylococcus Coriobacteriia.
8. A marker for detecting tubal edema induced by a chlamydia genital tract infection, wherein said marker is the type and abundance of the flora of claim 7.
9. Use of a method according to any one of claims 1-6 for studying the relationship between intestinal colonization and tubal edema following infection of the genital tract by chlamydia, for revealing the pathogenesis of tubal edema.
10. Use of a method of studying the relationship of chlamydia gut colonisation with tubal edema according to any one of claims 1 to 6 in the development of a diagnostic marker of chlamydia infection-induced tubal edema.

Description

Method for researching relationship between intestinal tract fixation and oviduct edema after chlamydia genital tract infection and application Technical Field The invention relates to the technical field of biological medicine, in particular to a method for researching the relationship between intestinal field planting and salpingemphraxis after chlamydia genital tract infection and application thereof. Background Genital tract infections are an important cause of female infertility and are also public health problems of general concern worldwide. The new cases of chlamydia trachomatis (CHLAMYDIA TRACHOMATIS, ct) genitourinary infections worldwide are about 1 million each year, which is a significant cause of acquired tubal infertility (Tubal Factor Infertility, TFI). Therefore, the method for clarifying the pathogenic mechanism of Ct genitourinary tract infection has important significance for breaking through the diagnosis and treatment bottleneck of female fallopian tube obstructive infertility. The study of the pathogenic mechanism of Ct depends on the establishment of a model of Ct animal infection, and at present, the establishment of a model of mouse genital tract infection by using Ct chlamydia (CHLAMYDIA MURIDARUM, cm) is a conventional animal model for studying the interaction of Ct with the host genital tract. The existing research shows that the chlamydia can spread to the intestinal tract after genital tract infection and realize the colonization, but the relation between the intestinal tract colonization of the chlamydia and the formation of persistent and stable tubal edema is not known at present, and a complete and standardized experimental method is lacking, which is used for systematically revealing the effect of the intestinal tract colonization of the chlamydia in the oviduct lesion caused by the chlamydia infection. Disclosure of Invention The invention aims to provide a method for researching the relation between intestinal tract field planting and salpingemphraxis after chlamydia genital tract infection and application thereof, which aims to solve the problem that a complete and standardized experimental method is lacking at present and is used for systematically researching the action and mechanism of the intestinal tract field planting in the salpingemphraxis caused by chlamydia genital tract infection, and the application of the method definitely plays a key role in the occurrence of the chlamydia genital tract infectious salpingemphraxis and the action thereof, thereby providing targets and experimental evidence for developing a diagnosis marker and/or prevention and intervention treatment of salpingemphraxis based on the intestinal flora. In order to achieve the above purpose, the invention provides a method for researching the relationship between intestinal tract field planting and oviduct edema after chlamydia genital tract infection, which comprises the following steps: S1, constructing a chlamydia genital tract infection animal model, dynamically monitoring chlamydia loading capacity of the animal model and analyzing the quantity and structure of intestinal flora; s2, constructing an animal model for the depletion of intestinal flora after chlamydia genital tract infection, and monitoring the fecal bacterial DNA content, the flora quantity, the weight change and the cecum condition of the animal model; S3, evaluating the influence of the depletion of intestinal flora on oviduct lesions induced by chlamydia genital tract infection; S4, constructing a chlamydia genital tract infection mouse model of fecal bacteria transplantation and remodeling, and detecting the number, weight and intestinal tract change condition of intestinal bacteria of the model animal; S5, evaluating whether intestinal flora can be rebuilt through fecal transplantation in a chlamydia genital tract infection animal model, and improving oviduct lesions; S6, screening marker species related to oviduct lesions induced by chlamydia genital tract infection. Preferably, in the step S1, a pair of differential chlamydia plasmid strains and a pair of differential chlamydia tc0668 strains are adopted to respectively construct a genital tract infection animal model of chlamydia, the chlamydia genital tract infection animal model is a mouse model, and the steps of constructing the chlamydia genital tract infection animal model are as follows: following adaptive feeding, depo days prior to infection, mice were vaccinated with the resuscitated strain via the vaginal route in synchronization with the estrus cycle of the mice; The plasmid differential chlamydia strain is a chlamydia murine plasmid carrying strain Cm NiggII and a chlamydia murine plasmid deletion strain CM PLASMID-free, and the tc0668 differential chlamydia strain is a chlamydia murine tc0668 wild strain Cm G13.32.1 and a chlamydia murine tc0668 mutant strain Cm G28.51.1; Dynamic monitoring of chlamydia load in animal model by collecting vaginal and rec