The Mechanism Of NLRP3 Inflammasome-induced Endothelial Injury In Pulmonary Arterial Hypertension

BACKGROUND:Pulmonary hypertension(PH)is a serious threat to human physical and mental health and expensive to treat;endothelial cell damage and dysfunction are its core pathological changes,and inflammatory factors are the key causative factors,NLRP3,as a key inflammatory factor,has an important role in the development of pulmonary hypertension disease,and the mechanism of action The mechanism of operation is still unclear.The Chinese medicine Chuanxiong has a long history and is widely used,and its extract Ligustrazine or Tetramethylpyrazine(TMP),which has the effects of scavenging oxygen free radicals,inhibiting inflammatory infiltration and regulating immunity,can inhibit NLRP3 expression and can improve pulmonary artery remodeling and fibrosis.This study intends to explore the mechanism of NLRP3 promoting endothelial injury in pulmonary hypertension and the mechanism of TMP preventing pulmonary hypertension by inhibiting NLRP3,to expand the pathological mechanism of pulmonary hypertension and to promote the application of traditional Chinese medicine in pulmonary hypertension.OBJECTIVE:1.to explore the interventional effects of NLRP3 on pulmonary artery pressure and right ventricular hypertrophy in rats with PH.2.to explore the effect of NLRP3 on the cellular behavior of rat pulmonary artery tissue.3.to explore the overall effect of NLRP3 on pulmonary circulation in rats with pulmonary hypertension.4.exploring the mechanisms of NLRP3 intervention in pulmonary hypertension.5.to explore the intervention effect of chuanxiongzine on NLRP3 and its mechanism.By exploring the above questions,the mechanism of NLRP3 inflammatory vesicles-induced pulmonary hypertension can be further clarified,providing targets and alternative drugs for the treatment of pulmonary hypertension.MATERIALS AND METHODS:NLRP3-/-rats were prepared using gene editing technology,and SD rats were taken as controls to induce PH model using Monocrotaline,and TMP intervention after successful modeling.Transthoracic puncture of the right ventricle was performed to assess hemodynamics,pulmonary artery casts and confocal techniques and methods were used to shape the 3D structure of pulmonary artery microcirculation,histology and TEM were used to observe the pathological changes and ultrastructure of pulmonary vessels,immunohistochemistry and Western blot were used to detect the expression and distribution of NLRP3,IL-1β,CASP-1,iNOS and PKG-1,and ELISA was used to analyze tissue Oxidative stress and other related indexes were analyzed by ELISA.RESULTS:1.Monocrotaline could successfully induce PH in rats,which showed pulmonary hypertension,right ventricular hypertrophy,capillary dilation,and significant reduction of small micro-arterial branches and microcirculation in PH rats.2.NLRP3 knockdown suppressed pulmonary artery pressure elevation,reduced right ventricular and pulmonary artery remodeling,reduced pulmonary artery microcirculation injury,endothelial endoplasmic reticulum and mitochondrial stress and pulmonary artery inflammatory infiltration and ROS levels,suppressed iNOS expression,and reduced PKG-1 expression inhibition in PH rats(P<0.05).3.TMP reduced pulmonary artery pressure,inhibited right ventricular and pulmonary artery remodeling,improved right ventricular and pulmonary artery fibrosis in PH rats;inhibited pulmonary artery inflammatory response and oxidative stress levels,suppressed NLRP3 inflammatory vesicle activation,promoted PKG-1 expression and inhibited iNOS(P<0.05).CONCLUSIONS:NLRP3 inflammatory vesicles can promote the development of pulmonary hypertension through the ROS-INOS-PKG1 signaling pathway.Chuanxiongzin has the effect of preventing pulmonary hypertension;the mechanism may be closely related to the inhibition of NLRP3,reduction of oxidative stress,inhibition of iNOS expression,and promotion of PKG-1 expression.