右美托咪定对脂多糖诱导的子宫内膜炎小鼠子宫组织中TLR4-NF-κB/NLRP3炎性小体介导的炎性反应的影响

Effect of Dexmedetomidine on Inflammatory Response Mediated by TLR4-NF-κB/NLRP3 Inflammatory Bodies in Uterine Tissue of Mice with Endometritis Induced by Lipopolysaccharide

目的探讨右美托咪定(DEX)对小鼠子宫内膜炎的作用及相关分子机制。方法将40只雌性C57BL/6小鼠随机分为对照组、模型组及右美托咪定组(10、20、40μg/kg),共计5组;腹腔注射给药30min后,采用小鼠子宫注射LPS溶液建立小鼠子宫内膜炎模型;24h后小鼠眼球取血,脱颈椎处死,解剖分离小鼠子宫组织;采用HE染色法检测小鼠子宫组织的病理变化并评分;采用ELISA法检测小鼠血清中IL-1β、IL-18、TNF-α的分泌水平;采用RT-PCR法检测小鼠子宫组织中IL-1β、IL-18、TNF-α的表达水平;采用Western blot法检测小鼠子宫组织中TLR4-NF-κB介导的NLRP3炎性小体活化相关分子的表达水平。结果HE染色结果表明与对照组小鼠比较,模型组小鼠的子宫组织中出现明显的炎性细胞浸润和肌层水肿及黏膜增生等炎性反应,给予右美托咪定能够抑制子宫内膜炎小鼠的子宫炎性细胞浸润及肌层水肿等炎性反应;ELISA结果表明与对照组小鼠比较,模型组小鼠血清中IL-1β、IL-18、TNF-α的含量显著增加,给予右美托咪定呈剂量依赖性降低子宫内膜炎小鼠血清中IL-1β、IL-18、TNF-α的含量;RT-PCR结果表明与对照组小鼠比较,模型组小鼠子宫组织中IL-1β、IL-18、TNF-αmRNA水平显著上调,给予右美托咪定呈剂量依赖性显著下调IL-1β、IL-18、TNF-αmRNA水平;Western blot法结果表明与对照组比较,模型组小鼠子宫组织中caspase-1、ASC、NLRP3、TLR4、p-p65/p65表达水平显著上调,给予右美托咪定能够显著抑制子宫内膜炎小鼠子宫组织中caspase-1、ASC、NLRP3、TLR4、p-p65/p65的表达。结论右美托咪定能够缓解脂多糖诱导的小鼠子宫内膜炎,并且与抑制子宫组织中TLR4-NF-κB/NLRP3炎性小体介导的炎性反应相关,对子宫内膜炎具有潜在的治疗作用。

Objective To investigate the effect of dexmedetomidine(DEX)on endometritis in mice and its related molecular mechanisms.Methods Forty female C57BL/6 mice were randomly divided into control group,model group and dexmedetomidine group(10,20,40μg/kg)for a total of 5 groups.After 30 minutes of intraperitoneal injection,a mouse model of endometritis was established by injecting LPS solution into the mouse uterus.After 24 hours,the mouse eyeballs were bled,the cervical vertebrae were sacrificed,and the mouse uterus tissue was dissected.The pathological changes of the mouse uterus were detected by HE staining and scored.The secretion levels of IL-1β,IL-18 and TNF-αin serum were detected by ELISA.The expression levels of IL-1β,IL-18 and TNF-αin mouse uterus were detected by RT-PCR.Western blot was used to detect the expression of TLR4-NF-κB-mediated NLRP3 inflammasome activation-related molecules in mouse uterus.Results The results of HE staining showed that compared with the control group,inflammatory cell infiltration and myometrial edema and mucosal hyperplasia were observed in the uterus of the model group,and dexmedetomidine was given to inhibit inflammatory reactions such as inflammatory cell infiltration and myometrial edema in mice.The results of ELISA showed that the levels of IL-1β,IL-18 and TNF-αin the serum of the model group were significantly increased compared with the control mice,and dexmedetomidine was administered in a dose-dependent manner to reduce the content of IL-1β,IL-18 and TNF-αin serum.The results of RT-PCR showed that IL-1β,IL-18 and TNF-αmRNA levels were significantly up-regulated in the uterus of the model group compared with the control group,and dexmedetomidine was administered to down-regulate IL-1β,IL-18,TNF-αmRNA levels in a dose-dependent manner.Western blot results showed that the expression levels of caspase-1,ASC,NLRP3,TLR4 and p-p65/p65 in the uterus of the model group were significantly up-regulated compared with the control group.The administration of dexmedetomidine significantly inhibited the expression of caspase-1,ASC,NLRP3,TLR4,p-p65/p65 in mouse uterus.Conclusion Dexmedetomidine has an anti-inflammatory effect,which associated with inhibiting TLR4-NF-κB/NLRP3 inflammasome,and has certain potential value in the treatment of endometritis.