基于ROS介导NLRP3炎症小体通路探讨蠲痛饮对子宫内膜异位症大鼠自噬的作用机制  

Exploring the mechanism of action of Juantong Drink on autophagy in rats with endome-triosis based on ROS-mediated NLRP3 inflammatory vesicle pathway

作　　者：刘骐语 李婧 孟凤云 刘英[2] 李卫红[1] LIU Qi-yu;LI Jing;MEMG Feng-yun;LIU Ying;LI Wei-hong(Guangxi University of Chinese Medicine,Nanning 530200,China;Ruikang Hospital Affiliated to Guan-gxi University of Chinese Medicine,Nanning 530010,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]广西中医药大学附属瑞康医院,广西南宁530010

出　　处：《时珍国医国药》2024年第9期2086-2091,共6页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81960884);广西自然科学基金(2023GXNSFAA026223,2020GXNSFAA238023);全国名中医陈慧侬学术思想与临床诊疗传承发展推广中心建设项目(桂中医大党〔2022〕23号)。

摘　　要：目的基于ROS介导NLRP3炎性小体通路探讨蠲痛饮对子宫内膜异位症自噬的作用机制。方法建立EMs大鼠模型,随机将72只SD大鼠分为假手术组、模型组、蠲痛饮(高、中、低)剂量组、地诺孕素组、氧化应激组,分别给予蠲痛饮高、中、低剂量(25.4,12.7,6.35g·kg^(-1))灌胃,地诺孕素组(0.21mg/kg)灌胃,氧化应激组,模型组和假手术组给予生理盐水(1ml/100g)灌胃,于取材前八天,氧化应激组通过腹腔注射10mg/kg BW Diquat建立氧化应激模型,对照组则注射等量生理盐水,之后饲养给药如常。各组大鼠连续用药28d。对各组大鼠光镜下异位内膜病理改变进行检测,DCFH-DA荧光探针检测内膜组织活性氧(ROS)水平,电镜观察内膜组织自噬小体数量;荧光定量PCR、免疫组化、蛋白免疫印迹法(Western blot)检测NLRP3、Caspase-1、LC3II、Beclin-1的mRNA及蛋白表达;酶联免疫吸附法(ELISA)检测血清中TNF-α的表达情况。结果与假手术组比较,模型组异位内膜ROS水平、NLRP3、Caspase-1蛋白及mRNA表达显著上调,Beclin-1、LC3B表达与自噬小体数量显著降低(P<0.05),血清TNF-α含量显著升高(P<0.05);与模型组对比,蠲痛饮高剂量组异位内膜ROS含量显著下调(P<0.05),自噬小体数量增加(P<0.05),蠲痛饮(高、中剂量组)、地诺孕素组NLRP3、Caspase-1蛋白及mRNA表达显著下调(P<0.05),血清TNF-α含量显著下调(P<0.05),蠲痛饮(高、中剂量组)、地诺孕素组LC3B、Beclin-1蛋白及mRNA表达均升高(P<0.05),其中蠲痛饮高剂量组升高最为显著(P<0.05),异位内膜组织病理形态明显改善。结论蠲痛饮能抑制异位内膜组织氧化应激反应,调控NLRP3炎性小体通路,激活自噬水平,促使异位内膜细胞凋亡,达到治疗EMs的目的。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,and LC3II,Beclin-1 protein and mRNA expression;enzyme-linked immunosorbent assay(ELISA)to detect the expression of TNF-αin 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-αcontent 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-αcontent was significantly down-regulated(P<0.05),and the expression of LC3B,Beclin-1 protein and mRNA was elevated in the Juan-tong Drink(high and medium dose

关 键 词：子宫内膜异位症 氧化应激 自噬 蠲痛饮 NLRP3炎症小体 

分 类 号：R285.5[医药卫生—中药学]