| The most common type of ectopic pregnancy in clinic is tubal ectopic pregnancy,and the treatment plan is mainly divided into expectant treatment,drug treatment and surgical treatment.The improvement of the level of early diagnosis makes it possible to use drug therapy for tubal ectopic pregnancy,especially for patients who are afraid of surgical treatment,have fertility requirements,and repeat tubal ectopic pregnancy.Nowadays,the effect of drug therapy is remarkable and the proportion of drug therapy for tubal ectopic pregnancy has increased significantly.In recent years,many clinical guidelines have confirmed the efficacy of drug therapy,especially Chinese medicine therapy for tubal ectopic pregnancy.Under the support of the national policy of accelerating the development of traditional Chinese medicine characteristics,it is of great significance to give better play to the characteristics of traditional Chinese medicine in the treatment of tubal ectopic pregnancy.Despite the continuous progress of detection methods,the accurate diagnosis of early pregnancy in unknown location is still a difficult problem.The pathogenesis of tubal ectopic pregnancy is still being explored continuously,and the mechanism research of Traditional Chinese medicine preparation for tubal ectopic pregnancy treatment is also being carried out comprehensively.Professor Deng Gaopi and his research group have carried out basic and clinical research on tubal ectopic pregnancy for more than 20 years.Huayu Xiaozheng formula has been clinically verified in many centers with significant efficacy,and there are also abundant data in basic research on Huayu Xiaozheng formula.Based on this,huayu Xiaozheng granule is the first Chinese medicine preparation for the adjuvant treatment of early tubal ectopic pregnancy,which is specialized in killing embryos and eliminating diseases.Huayu Xiaozheng granules have fill the blank of traditional Chinese medicine preparation of clinical targeted treatment of tubal ectopic pregnancy,its formulation is simplified,dosage form is stable and quality is controllable.Based on the above background,retrospective analysis was conducted on the early tubal ectopic pregnancy patients hospitalized in the Women and Children’s Center of the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine in recent 5 years who chose drug treatment,to observe the clinical efficacy of Huayu Xiaozheng granules in the treatment of early tubal ectopic pregnancy,and analyze the relevant factors affecting the clinical efficacy of Huayu Xiaozheng granules.At the same time,clinical samples were collected to verify the expression of Talin1 protein in tubal ectopic pregnancy tissues and villi.On the basis of literature support and clinical validation,using HTR-8/SVneo cell line as the carrier,the regulation of Talin1 protein on the invasion and migration of gestational trophoblast cells was verified.Finally,based on the previous work of the research group,the mechanism of Huayu Xiaozheng granules treating early tubal ectopic pregnancy was further explored from the perspective of Talin1 regulating the invasion of pregnancy trophoblast cells.1.Clinical researchObjectiveTo observe the clinical efficacy of Huayu Xiaozheng granules in the treatment of early tubal ectopic pregnancy and analyze the related factors affecting the clinical efficacy of Huayu Xiaozheng granules.MethodsA total of 374 tubal ectopic pregnancy patients who were hospitalized in the First Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2016 to March 2021 and chose drug therapy were included in the study.All patients were treated with integrated Traditional Chinese and Western medicine for tubal ectopic pregnancy.According to whether Huayu Xiaozheng granules were used or not,they were divided into Huayu Xiaozheng granules group and Xuefu Zhuyu granules group.The serum β-HCG value,the maximum diameter line value of pelvic mass and the maximum diameter line value of pelvic fluid were compared between the two groups before and after treatment,and the clinical efficacy was analyzed.We used univariate and multivariate logistic regression analysis to explore the possible related factors affecting the clinical efficacy of Huayu Xiaozheng granules.Results(1)The total effective rate of Huayu Xiaozheng granule group was 93.7%,and the total effective rate of Xuefu Zhuyu granule group was 82.7%,the curative effect of Huayu Xiaozheng granule in treating tubal ectopic pregnancy is better than Xuefu Zhuyu granule group(P<0.01).(2)Compared with before treatment,β-HCG value in 2 groups decreased significantly on day 7 of treatment,with statistical significance(P<0.01).After treatment,the maximum diameter line of pelvic mass and the maximum diameter line of pelvic effusion in Huayu Xiaozheng granule group were significantly smaller than those before treatment,and the difference was statistically significant(P<0.01).(3)The cutoff value of serum β-HCG before treatment is 1166.5 IU/L,AUC is 85.4%,specificity is 98%,sensitivity is 60%.It has certain predictive value for the treatment outcome of Huayu Xiaozheng granules.Conclusion(1)Huayu Xiaozheng granules can improve the clinical effective rate of early tubal ectopic pregnancy patients,and have obvious advantages in reducing β-HCG value and reducing the maximum diameter line of pelvic mass and effusion,with good safety and application value.(2)The serum β-HCG value before treatment was independent risk factors affecting the curative effect of Huayu Xiaozheng granules on tubal ectopic pregnancy.(3)The serum β-HCG value higher than 1166.5 IU/L is the clinical decision point for treatment.2.Experimental research(1)Differential expression of Talin1 between gestational fallopian tube tissue and villi and verification of invasion regulation of gestational trophoblast cellsObjectiveTo verify the expression difference of Talinl in the gestational fallopian tube tissue and the gestational villi of the fallopian tube and its effect on the invasion and migration of HTR-8/SVneo cells,and to explore its mechanism.Methods①We collected 15 cases of normal fallopian tube tissue,15 cases of gestational fallopian tube tissue and gestational fallopian tube villi,and 15 cases of normal intrauterine pregnancy villi from April 2020 to December 2020 in the Women and Children’s Center of the First Affiliated Hospital of Guangzhou University of Chinese Medicine.Western Blot and Immunohistochemistry were used to detect the different expression of Talinl in normal human fallopian tubes and gestational fallopian tubes,and the differences in protein expression between human intrauterine and tubal ectopic pregnancy villi.②Talin1 siRNA was transfected in HTR-8/SVneo cells,the effect of Talinl on invasion and migration of HTR-8/SVneo cells was detected by scratch assay and Transwell assay.③The expressions of MMP-2,MMP-9,N-cadherin and Snail in HTR-8/SVneo cells after si-Talin1 were detected by qRT-PCR and Western Blot.Results①Positive expression of Talin1 can be detected in normal and pregnant fallopian tube tissues,mainly expressed in the cytoplasm of ciliated cells.②The protein expression of Talinl in pregnant fallopian tube tissue was higher than that in normal fallopian tube tissue(P<0.01),Talin1 expression in tubal ectopic pregnancy villi was higher than that in intrauterine pregnancy villi(P<0.01).③Si-Talinl inhibits the invasion and migration of HTR-8/SVneo cells.Compared with the control group,the migration distance of cells in si-Talin1 group was reduced,and the change of scratch area was small,showing statistical differences(P<0.05),the number of transmembrane cells in si-Talin1 group was decreased,with statistical difference(P<0.01).④Talin1 regulates the expression of invasion-related factors N-cadherin,MMP-2,Snail and MMP-9 in HTR-8/SVneo cells,compared with the control group,after transfection of Talin1 siRNA into HTR-8/SVneo cells,the expression levels of N-cadherin,MMP-2 and Snail in HTR-8/SVneo cells had been reduced significantly(P<0.05),while the expression levels of MMP-9 were significantly up-regulated(P<0.05).Conclusion①The expression of Talinl in tubal tissue and villi of tubal ectopic pregnancy is significantly increased,which may be a potential target for diagnosis and treatment of early tubal ectopic pregnancy.②Talin1 silencing inhibits invasion and migration of human chorionic trophoblast cells HTR-8/SVneo,the mechanism of which may be closely related to Talin1 regulation related invasion factors.The regulation of Talin1 on the invasion of gestational trophoblast may be one of the mechanisms of tubal ectopic pregnancy.(2)Study on the mechanism of Huayu Xiaozheng Granules containing drug serum interfering with the invasion of pregnancy trophoblast cells and the regulation of Talin1 expressionObjectiveTo study the effects of Huayu Xiaozheng granules containing serum on the invasion,proliferation and migration of HTR-8/SVneo cells,and to explore its mechanism.Methods①Preparation of drug-containing serum,CCK8 assay was used to detect the effects of different concentrations and doses of Huayu Xiaozheng granules containing serum on the proliferation of HTR-8/SVneo cells.②Scratch test and Transwell test were used to detect the effects of Huayu Xiaozheng granule containing serum on the invasion and migration of HTR-8/SVneo cells.③The expression of Talin1,MMP-2,N-cadherin,Snail,Rap1GAP and ITGB3 pathway proteins in HTR-8/SVneo cells were detected by qRT-PCR and Western Blot.Results①Compared with blank control group,the 50%concentration of Huayu Xiaozheng granules containing serum in high and low dose groups had no significant effect on the survival rate of HTR-8/SVneo cells(P>0.05).The survival rate of HTR-8/SVneo cells was significantly decreased in both the 10%and 30%Huayu Xiaozheng granule containing serum high-dose groups(P<0.05,P<0.01),the inhibition of cell proliferation was more significant in the 10%Huayu Xiaozheng granule containing serum high-dose group(P<0.05).②Compared with the blank control group,after 48h of intervention,the migration distance of cells in the 10%Huayu Xiaozheng granule containing serum high-dose group decreased and the change of scratch area was small,with statistically significant differences(P<0.01),there was no statistical difference compared with MTX group(P>0.05).Compared with the blank control group,10%Huayu Xiaozheng granule containing serum high-dose group could inhibit the invasion of HTR-8/SVneo cells,and the difference was statistically significant(P<0.01),compared with MTX group,the number of transmembrane cells in 10%Huayu Xiaozheng granule containing serum high-dose group was significantly reduced,and the difference was statistically significant(P<0.01).③Compared with blank control group,Huayu Xiaoshen granules containing serum down-regulated the expressions of Talinl and invasion related factors MMP-2,N-cadherin and Snail in HTR-8/SVneo cells(P<0.05,P<0.01).④Compared with blank control group,the expression of Rap 1 GAP and ITGB3 pathway proteins were down-regulated in Huayu Xiaozheng granules containing serum(P<0.05,P<0.01).Conclusion①Talinl may be a potential target for Huayu Xiaozheng granules containing serum to inhibit the invasion of pregnancy trophoblast cells.②Huayu Xiaozheng granules containing serum may inhibit the invasion and migration of gestational trophoblast cells by down-regulating Talin1 protein and related invasive proteins,its mechanism may be related to the regulation of Rapl/Talin1/ITGB3 pathway.③Huayu Xiaozheng granules can inhibit the invasion of pregnancy villi regulated by Talin1 to the oviduct tissue,causes the collaterals stasis to remove and truncated zygote abnormal planting,thus play the role of embryo killing in clinical application... |
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