摘要
目的:研究硫氧还蛋白相互作用蛋白∕NOD样受体热蛋白结构域相关蛋白3(TXNIP∕NLRP3)信号通路对多囊卵巢综合征(PCOS)-胰岛素抵抗(IR)大鼠卵巢慢性炎症的影响。方法:颈背部皮下注射脱氢表雄酮(DHEA)溶液0.2 ml(6 mg/100 g)建立PCOS-IR大鼠模型,建立成功的模型大鼠随机分为模型组、siRNA NC组、TXNIP siRNA组、RES干预组,每组8只,正常组8只。siRNA NC组、TXNIP siRNA组分别颈背部皮下注射5 nmol siRNA NC、TXNIP siRNA,5 d/次;RES干预组颈背部皮下注射50 mg/kg RES,正常组和模型组颈背部皮下注射等体积生理盐水,1次/d,10 d。放射免疫试剂盒检测血清中睾酮(T)、卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E_(2))、胰岛素样生长因子-1(IGF-1)水平,血糖仪检测血清中空腹血糖(FBG)水平,ELISA检测血清中空腹胰岛素(FINS)、IL-6、IL-1β、TNF-α水平,并计算胰岛素抵抗指数(HOMA-IR)、胰岛素敏感指数(ISI)。HE染色观察卵巢卵泡发育情况;qRT-PCR和Western blot检测卵巢组织中TXNIP、NLRP3 mRNA和蛋白,半胱氨酸天冬氨酸蛋白水解酶1(caspase-1)、pro-caspase-1蛋白水平。结果:模型组和siRNA NC组卵泡腔变大、出现囊肿现象,颗粒细胞层减少;TXNIP siRNA组和RES干预组出现黄体,卵泡腔变小,颗粒层增厚。与正常组相比,模型组、siRNA NC组T、LH、E_(2)、IGF-1、FBG、FINS、HOMA-IR,IL-6、IL-1β、TNF-α水平,卵巢组织中TXNIP、NLRP3 mRNA和蛋白水平,pro-caspase-1/caspase-1蛋白水平升高,FSH水平降低(P<0.05);分别与模型组、siRNA NC组相比,TXNIP siRNA组、RES干预组T、LH、E_(2)、IGF-1、FBG、FINS、HOMA-IR,IL-6、IL-1β、TNF-α水平,卵巢组织中TXNIP、NLRP3 mRNA和蛋白水平,pro-caspase-1/caspase-1蛋白水平降低,FSH水平升高(P<0.05)。结论:抑制TXNIP/NLRP3信号通路可实现对PCOS-IR大鼠卵巢慢性炎症的缓解。
Objective:To investigate the effect of thioredoxin-interacting protein/NOD-like receptor pyrin domain-containing protein 3(TXNIP/NLRP3)signaling pathway on chronic ovarian inflammation in rats with polycystic ovary syndrome(PCOS)-insulin resistance(IR).Methods:The PCOS-IR rat model was established by subcutaneous injection of dehydroepiandrosterone(DHEA)solution 0.2 ml(6 mg/100 g).The rats were randomly divided into model group,siRNA NC group,TXNIP siRNA group and RES intervention group,with 8 rats in each group,and 8 rats in the normal group.In siRNA NC group and TXNIP siRNA group,5 nmol siRNA NC and TXNIP siRNA were injected subcutaneously into the back of neck,once every 5 days;in the RES intervention group,50 mg/kg RES was injected subcutaneously into the neck and back,the normal group and the model group were subcutaneously injected with the same volume of normal saline,once a day,for 10 days.The levels of testosterone(T),follicle stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E_(2))and insulin-like growth factor-1(IGF-1)were detected by radioimmunoassay kit,the level of fasting blood glucose(FBG)was measured by blood glucose meter,the levels of fasting insulin(FINS),IL-6,IL-1β,TNF-αwere detected by ELISA,and insulin resistance index(HOMA-IR)and insulin sensitivity index(ISI)were calculated.Hematoxylin eosin(HE)staining was used to observe the development of ovarian follicles;the mRNA and protein levels of TXNIP and NLRP3,cysteinyl aspartate specific proteinase1(caspase-1)protein levels were detected by qRT-PCR and Western blot.Results:In the model group and siRNA NC group,the follicle cavity became larger and cysts appeared,and the granular cell layer decreased;in TXNIP siRNA group and RES intervention group,corpus luteum appeared,follicle cavity became smaller and granular layer thickened.Compared with normal group,the levels of T,LH,E_(2),IGF-1,FBG,FINS,HOMA-IR,IL-6,IL-1β,TNF-α,the mRNA and protein levels of TXNIP and NLRP3,pro-caspase-1/caspase-1 protein level in ovarian tissue were higher in model group and siRNA NC group,and the level of FSH was lower(P<0.05);compared with model group and siRNA NC group,the levels of T,LH,E_(2),IGF-1,FBG,FINS,HOMA-IR,IL-6,IL-1β,TNF-α,the mRNA and protein levels of TXNIP and NLRP3,pro-caspase-1/caspase-1 protein level in TXNIP siRNA group and RES intervention group were lower,and the level of FSH was higher(P<0.05).Conclusion:Inhibition of TXNIP/NLRP3 signaling pathway can alleviate the chronic inflammation of ovary in PCOS-IR rats and alleviate the disease.
作者
孙文萍
赵得雄
王榀华
王生兰
李洁
康燕
SUN Wenping;ZHAO Dexiong;WANG Zuhua;WANG Shenglan;LI Jie;KANG Yan(Department of Obstetrics and Gynecology,Qinghai Red Cross Hospital,Qinghai 810000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第11期1300-1305,共6页
Chinese Journal of Immunology
基金
青海省卫生健康委员会指导性课题(2019-wjzdx-69)。
