原儿茶酸对LPS诱导的小鼠子宫内膜炎的作用机制研究

doi:10.16431/j.cnki.1671-7236.2022.04.041

摘要/Abstract

摘要： 【目的】探究原儿茶酸对脂多糖(LPS)诱导的小鼠子宫内膜炎的作用机制。【方法】将60只小鼠随机分为对照组、LPS组、LPS+不同浓度原儿茶酸(20、40、80 mg/kg)组共5组,每组各12只。对照组小鼠用微量注射器经阴道灌注50 mL生理盐水,LPS组灌注50 mL LPS(1 mg/kg),原儿茶酸治疗组灌注50 mL LPS前1 h分别腹腔注射20、40、80 mg/kg原儿茶酸,注射24 h后,颈椎脱臼法处死所有小鼠。收集子宫组织,通过HE染色检测子宫组织病理变化,用试剂盒检测髓过氧化物酶(MPO)活性,ELISA法检测炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和IL-6含量;Western blotting法检测p65、核因子-κB(NF-κB)的抑制蛋白(IκB)以及磷酸化p65、IκB(p-p65、p-IκB)蛋白表达水平。【结果】组织病理结果显示,对照组小鼠子宫组织形态正常,子宫内膜上皮结构正常;LPS组小鼠子宫组织出现严重的炎性细胞浸润和子宫内膜上皮充血及水肿。与LPS组相比,随着原儿茶酸浓度的增加,小鼠子宫组织中炎性细胞明显减少,子宫内膜上皮充血水肿情况也明显得到改善。MPO活性检测表明,与对照组相比,LPS组MPO活性极显著升高(P<0.01);与LPS组相比,原儿茶酸治疗组MPO活性均极显著降低(P<0.01),并呈剂量依赖性。ELISA结果表明,与对照组相比,LPS组小鼠子宫内膜组织中TNF-α、IL-1β和IL-6含量均极显著升高(P<0.01);与LPS组相比,原儿茶酸治疗组小鼠子宫内膜组织中炎性细胞因子TNF-α、IL-1β和IL-6含量均极显著降低(P<0.01),并呈剂量依赖性。Western blotting结果表明,与对照组相比,LPS组p-p65和p-IκB的表达水平均极显著升高(P<0.01);与LPS组相比,原儿茶酸治疗组p-p65和p-IκB的表达水平均极显著降低(P<0.01),且呈剂量依赖性。【结论】原儿茶酸通过减轻子宫组织的病理变化,降低子宫组织MPO活性,抑制NF-κB信号通路及炎症细胞因子的产生,对LPS诱导的小鼠子宫内膜炎起到保护作用。

关键词: 原儿茶酸; 脂多糖(LPS); 子宫内膜炎; NF-κB信号通路

Abstract: 【Objective】 This study was aimed to investigate the protective effect and mechanism of protocatechuic acid on lipopolysaccharide (LPS) induced endometritis in mice.【Method】 Sixty mice were randomly divided into control group,LPS group and LPS+ procatechuic acid (20,40,80 mg/kg) groups,with 12 mice in each group.The control group was injected with 50 mL normal saline via microsyringe,the LPS group was injected with 50 mL LPS (1 mg/kg),and the protocatechuic acid treatment groups were injected intraperitoneally with 20,40 and 80 mg/kg protocatechuic acid 1 h before the injection of 50 mL LPS.After 24 h of injection,all the mice were sacrificed by cervical dislocation.The uterus tissues were collected,and the pathological changes of the uterus tissues were detected by HE staining,myeloperoxidase (MPO) activity was detected by kit,and the contents of inflammatory factors tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β) and IL-6 were detected by ELISA.The expression levels of p65 and NF-κB inhibitor protein (IκB) and phosphorylaed p65 and IκB protein (p-p65 and p-IκB) were detected by Western blotting.【Result】 Histopathological results showed that the endometrial epithelial structure of mice in control group was normal.The uterus tissues of mice in LPS group showed severe inflammatory cell infiltration and endometrial epithelial hyperemia and edema.Compared with LPS group,with the increase of protocatechuic acid concentration,the inflammatory cells in the uterus tissues of mice were significantly reduced,and the hyperemia and edema of endometrial epithelium were significantly improved.The activity of MPO in LPS group was extremely significantly higher than that in control group (P<0.01).Compared with LPS group,MPO activity in protocatechuic acid treatment group was extremely significantly decreased in a dose-dependent manner (P<0.01).ELISA results showed that compared with control group,the contents of TNF-α,IL-1β and IL-6 in endometrial tissues of mice in LPS group were extremely significantly increased (P<0.01).Compared with LPS group,the contents of inflammatory cytokines TNF-α,IL-1β and IL-6 in endometrial tissues of mice treated with protocatechuic acid were extremely significantly decreased (P<0.01) in a dose-dependent manner.Western blotting results showed that the expression of p-p65 and p-IκB in LPS group were extremely significantly increased compared with control group (P<0.01).Compared with LPS group,the expression of p-p65 and p-IκB in protocatechuic acid treatment group were extremely significantly decreased in a dose-dependent manner (P<0.01).【Conclusion】 Protocatechuic acid played a protective effect on lPS-induced endometritis in mice by alleviating pathological changes,decreasing MPO activity,inhibiting NF-κB signaling pathway and the production of inflammatory cytokines.

Key words: protocatechuic acid; lipopolysaccharide (LPS); endometritis; NF-κB signaling pathway

中图分类号:

S857.4

张玉珠, 于超, 郑慧华, 唐欣悦, 黄荣磊, 谢光洪. 原儿茶酸对LPS诱导的小鼠子宫内膜炎的作用机制研究[J]. 中国畜牧兽医, 2022, 49(4): 1585-1591.

ZHANG Yuzhu, YU Chao, ZHENG Huihua, TANG Xinyue, HUANG Ronglei, XIE Guanghong. Mechanism Study of Protocatechuic Acid on LPS-induced Endometritis in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1585-1591.

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