摘要
目的:探讨microRNA-703(miR-703)调控NLRP3介导的焦亡和炎症反应影响小鼠子宫内膜异位症(EM)进展机制。方法:采用小鼠-小鼠腹腔植入方法构建子宫内膜异位症BABL/C小鼠模型。将小鼠分为EM组和对照组。免疫印迹法和酶联免疫法检测两组焦亡标志蛋白(NLRP3、pro-caspase-1、caspase-1)和炎症因子表达(IL-1β、IL-18、TNF-α、IL-6)。实时荧光定量PCR检测两组miR-703含量。造模成功的小鼠分别腹腔注射miR-703 NC(EM+NC阴性组)、pre-miR-703(EM+miR-703组),检测两组小鼠子宫内膜异位组织大小,焦亡标志蛋白及炎性因子表达。结果:EM组小鼠NLRP3、caspase-1蛋白表达均高于对照组,炎症因子IL-1β、IL-18、TNF-α、IL-6水平(1.33±0.11ng/ml,0.66±0.08ng/ml,179.69±3.78pg/ml,90.375±4.63pg/ml)均高于对照组(0.30±0.05ng/ml,0.21±0.03ng/ml,50.51±2.96pg/ml,24.99±1.81pg/ml),异位组织中miR-703(0.52±0.09)低于对照组(1.00±0.07)(均P<0.05);在EM小鼠体内转染pre-miR-703后,EM+miR-703组EM组织变小,miR-703表达高于EM+NC阴性组,细胞焦亡标志蛋白表达低于EM+NC阴性组,炎症因子水平均低于EM+NC阴性组(均P<0.05)。结论:miR-703可能通过抑制NLRP3介导的焦亡和炎症反应改善EM症状。
Objective:To investigate the mechanism progression of the influence of NLRP3-mediated pyroptosis and inflammatory response regulated by microRNA-703(miR-703)of mice on their endometriosis(EM).Methods:The BABL/C mouse model of endometriosis was established by mouse-to-mouse intraperitoneal implantation.Mice were divided into study group and control group.Immunoblotting and enzyme-linked immunosorbent assay were used to detect the pyroptosis marker proteins(NLRP3,pro-caspase-1,and caspase-1)and the inflammatory factors expressions(IL-1β,IL-18,TNF-αand IL-6).Real-time PCR was used to detect the content of miR-703 in the two groups.Mice that were successfully modeled were intraperitoneally injected with miR-703 NC(EM+NC negative group)and pre-miR-703(EM+miR-703 group),respectively.The size of the endometriotic tissue and the expressions of pyroptosis marker proteins and inflammatory factors of the mice in the two groups were detected.Results:The protein expressions of NLRP3 and caspase-1 of the mice in the study group were significantly higher than those of the mice in the control group.The levels of inflammatory cytokines,such as IL-1β,IL-18,TNF-α,and IL-6,of the mice in the study group were 1.33±0.11ng/ml,0.66±0.08ng/ml,179.69±3.78pg/ml,and 90.375±4.63pg/ml,which were significantly higher than those group(0.30±0.05ng/ml,0.21±0.03ng/ml,50.51±2.96pg/ml,and 24.99±1.81pg/ml)of the mice in the control group.The expression of miR-703(0.52±0.09)in the ectopic tissues of the mice in the study group was significantly lower than that(1.00±0.07)of the mice in the control group(all P<0.05).After transfection of pre-miR-703 in the mice with EM,the EM tissues of the mice in EM+miR-703 group became smaller,the expression of miR-703of the mice in EM+miR-703group was significantly higher than that of the mice in EM+NC negative group,the expression of pyroptosis marker protein of the mice in EM+miR-703group was significantly lower than that of the mice in EM+NC negative group,and the levels of inflammatory factors of the mice in EM+miR-703group were significantly lower than those of the mice in EM+NC negative group(all P<0.05).Conclusion:MiR-703may improve endometriosis symptoms by inhibiting NLRP3-mediated pyroptosis and inflammatory response.
作者
张玲云
何佳英
王辰玥
徐蓓蕾
ZHANG Lingyun;HE Jiaying;WANG Chenyue;XU Beilei(Dingqiao Branch,Hangzhou Hospital of Traditional Chinese Medicine,Zhejiang Province,310044)
出处
《中国计划生育学杂志》
2023年第1期15-18,共4页
Chinese Journal of Family Planning
基金
杭州市科技局杭州市农业与社会发展科研项目(20201203b204)。
