| Objective: To investigate the expressions of NLRP3(NOD-like receptor protein3),Caspase-1(cycsteinyl aspartate-specific protease-1)and interleukin-1β(IL-1β)in villous tissues of patients with missed abortion and normal early pregnant women.And to analyze the expression differences of NLRP3,Caspase-1 and IL-1β in the two groups of villous tissues,to further explore the impact of NLRP3/Caspase-1/IL-1β inflammasome pathway on the occurrence and development of missed abortion,so as to provide the certain theoretical basis for the prevention and treatment of missed abortion.Methods:Twenty-five patients who were diagnosed with miscarriage at the Pingguo People’s Hospital and the Pingguo Maternal and Child Health Hospital in Guangxi Zhuang Autonomous Region from April 2019 to September 2019 were selected as the study group,and twenty-five normal early pregnant women were the control group.Collected fresh villi tissue immediately after induced abortion.Real-time quantitative PCR technology was used to detect the m RNA expression of NLRP3,Caspase-1 and IL-1β in the two groups.Pathological sections were prepared from two groups of villous tissues,and the pathological sections were treated with Hematoxylin-Eosin staining.The morphological structure of the two groups of villi were observed under an optical microscope.Immunohistochemistry streptavidin-perosidase was used to detect expressions and localizations of NLRP3,Caspase-1,and IL-1β.After taking photos,analyze the photos with Image Pro Plus 6.0 software to calculate the relative expression levels of NLRP3,Caspase-1 and IL-1β.Results: 1.Real-time quantitative PCR results: the relative expression of NLRP3,Caspase-1 and IL-1β m RNA in the villi tissue of the study group was significantly higher than that of the normal control group(P <0.05),the difference was statistically significant.2.After HE staining,observation under the light microscope showed that the hairs of the control group had regular morphology,consistent size,complete hair structure and covered with two layers of trophoblast cells.The outer layers were syncytial trophoblast whose cytoplasm was fine or empty vesicular,and the trophoblastic cells have complete structure and were arranged neatly.The inner layer were cell trophoblastic layers,which was cubic,polygonal,or oval,with clear boundaries,transparent cytoplasm,and no edema in the villous stroma.The normal morphological structure of the villi in the study group was changed,with varying degrees of degenerative lesions or even necrosis,disordered trophoblast arrangement,reduced or even disappeared structure of the two layers of trophoblasts,reduced villous stroma,mesenchymal edema or fibrosis.Old blood clots and infiltration of inflammatory cells could be seen.3.Observation of the villi samples treated by immunohistochemical SP method with a light microscope showed that the villous trophoblast cells of the control group and the study group expressed NLRP3,Caspase-1 and IL-1β proteins,which were mainly expressed in the cytoplasm of trophoblast cells.There is a small amount of expression in the interstitial villi,and the staining intensity in the study group is significantly higher than that in the control group.After taking a picture,it was analyzed by Image Pro Plus 6.0 software,and AOD(average optical density,average optical density value)was used to represent the intensity of staining.The results showed that the expression level of NLRP3,Caspase-1 and IL-1βprotein in the study group was significantly higher than that in the control group,and there was a statistically significant difference between the two(P <0.05).4.The results of Pearson correlation analysis showed that in the villi tissue of the study group,relative expression of NLRP3 protein was positively correlated with the relative expression of Caspase-1 protein,and the relative expression of NLRP3 protein was positively correlated with the relative expression of IL-1βprotein.There was also a positive correlation between the relative expression of Caspase-1 protein and the relative expression of IL-1β protein.In the villi tissue of the control group,the relative expression of NLRP3 protein was not correlated with the relative expression of Caspase-1 protein,and the relative expression of NLRP3 protein was not relative to IL-1β protein.The relative expression level of Caspase-1 protein and the relative expression level of IL-1βprotein was positively correlated.Conclusions: 1.The activation of the NLRP3/Caspase-1/IL-1β inflammasome pathway is activated in the villous tissue of patients with missed abortion,and plays a role in the occurrence and development of missed abortion.2.The NLRP3/Caspase-1/IL-1β inflammasome pathway is not activated in normal early pregnancy villous tissue.Caspase-1 and IL-1β play roles in maintaining normal pregnancy through other pathways. |
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