Exploring the mechanism of action of Juantong Drink on autophagy in rats with endome-triosis based on ROS-mediated NLRP3 inflammatory vesicle pathway
Objective To investigate the mechanism of Juantong Drink on autophagy in endometriosis rats based on ROS-media-ted NLRP3 inflammatory vesicle pathway.Methods Establishment of the EMS rat model,and 72 SD rats were randomly divided into the sham operation group,model group,Juantong Drink(high,medium and low)dose group,Dinorelgestrol group,oxida-tive stress group and oxidative stress+high dose group,and the oxidative stress and oxidative stress+high dose groups were injec-ted intraperitoneally with 10mg/kg body The oxidative stress and oxidative stress+high dose groups were injected intraperitoneal-ly with 10mg/kg body weight Diquat to establish the oxidative stress model,while the control group was injected with an equal a-mount of saline,and then fed as usual.Each group was administered the drug continuously for 28 d.The pathological changes of ectopic endothelium in each group were examined by light microscopy,DCFH-DA fluorescent probe to detect the level of reactive oxygen species(ROS)in endothelial tissues;Fluorescent PCR,immunohistochemistry,and protein immunoblotting(Western blot)to detect the proteins and proteins of NLRP3,Caspase-1,andLC3Ⅱ,Beclin-1 protein and mRNA expression;enzyme-linked immunosorbent assay(ELISA)to detect the expression of TNF-α in serum.Results Compared with the sham-operated group,the ectopic endothelial ROS level,NLRP3,Caspase-1 protein and mRNA expression were significantly up-regulated in the model group,and the expression of Beclin-1,LC3Bwere significantly reduced(P＜0.05),and serum TNF-α content was significantly increased(P＜0.05);compared with the model group,ectopic endothelial ROS content was significantly down-regulated in the Juantong Drink high-dose group and high-dose+oxidative stress group(P＜0.05),NLRP3,Caspase-1 protein and mRNA expression was significantly down-regulated in the dienogest group(P＜0.05),serum TNF-α content was significantly down-regulated(P＜0.05),and the expression of LC3 B,Beclin-1 protein and mRNA was elevated in the Juan-tong Drink(high and medium dose groups),dienogest group(The expression of LC3B,Beclin-1 protein and mRNA were all increased in Juantong Drink(high and medium dose groups)and Dinorelgestrol group(P＜0.05),and the histopathological morphology of ectopic endothelium was significantly improved in Juantong Drink(high dose group).Conclusion Juantong Drink can inhibit oxidative stress in ectopic endothelial tissue,regulate NLRP3 inflammatory vesicle pathway,activate autophagy level,and promote apoptosis of ectopic endothelial cells to achieve the purpose of treating EMs.
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