目的:子宫内膜炎的细菌性致病状况在子宫内膜发展,影响人和动物。褐孔菌多糖(IOP),一种活生生的黑孔菌鸡尾酒,已被证明具有相对广泛的生物活性,并且可以在各种疾病中发挥作用。然而,从目前报道的文章来看,没有关于IPO在脂多糖(LPS)诱导的子宫内膜炎症状中的抗炎作用的信息。因此,本研究仔细观察了眼压对LPS诱导小鼠子宫内膜炎症状的影响,阐明了IOP对身体的保护机制,阐明了眼压的潜在机制。
方法:将72只BALB/c雌性实验小鼠分为几组进行比较。他们是空白对照组,LPS组,LPS+IOP组(还探讨了IOP剂量对小鼠的影响,分为低,中等,和高)和LPS+阿莫西林组。除对照组外,所有组均向子宫内注入LPS。LPS+IOP组和LPS+阿莫西林组的小鼠在LPS攻击3小时后口服施用IOP或阿莫西林。组织病理学和髓过氧化物酶(MPO)活性用于检测子宫组织损伤,和细胞因子水平用于测量子宫炎症。观察Toll样受体4/核因子-κB(TLR4/NF-κB)相关蛋白在炎症信号通路中的表达。
结果:病理和MPO活性分析显示,眼压减轻了LPS诱导的子宫组织损伤。采用定量逆转录-聚合酶链反应检测和定量研究小鼠细胞的RNA信息,具有较高的准确度和灵敏度。从测试结果来看,眼压确实能有效控制促炎细胞因子如白细胞介素-6(IL-6)的释放,IL-1β,IL-8和肿瘤坏死因子-α(TNF-α),避免身体的免疫反应。对子宫组织细胞成分的分析也证实了炎症介质诱导的一氧化氮合酶(iNOS)的表达水平也大大降低。细胞合成的免疫印迹分析结果显示,眼压主要抑制体内TLR4和髓样分化因子88的蛋白表达。
结论:本研究证明眼压的作用机制是通过抑制TLR4/NF-κB信号通路,减少机体细胞内促炎细胞因子的释放,从而减轻LPS诱导的子宫内膜炎的症状。因此,眼压可能是预防和治疗LPS诱导的子宫内膜炎的有效药物。
OBJECTIVE: Endometritis bacterial pathogenic condition that affects both humans and animals develops in the inner lining of the uterus. Inonotus obliquus polysaccharide (IOP), an active cocktail of Inonotus obliquus, has been shown to have a relatively wide range of biological activities and can play a role in various diseases. However, from the currently reported article, there is no information about the anti-inflammatory effect of IPO in the symptoms of lipopolysaccharide (LPS)-induced endometritis. Therefore, this study carefully observed the phenomenon of IOP on the symptoms of endometritis induced by LPS in mice, elucidated the protective mechanism of IOP on the body, and clarified the potential mechanism of IOP.
METHODS: A total of 72 BALB/c female experimental mice were divided into several groups for comparison. They were the blank control group, the LPS group, the LPS+ IOP group (the effect of IOP dose on mice was also explored, divided into low, medium, and high) and LPS+ amoxicillin group. All groups except control group were infused with LPS into the uterus. The mice of LPS+ IOP groups and LPS+ amoxicillin group were orally administered with IOP or amoxicillin after LPS challenge for 3 hours. Histopathology and myeloperoxidase (MPO) activity were used to detect uterine tissue injury, and cytokine levels were used to measure uterine inflammation. The expression of toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-κB)-related proteins in the inflammatory signaling pathway was observed.
RESULTS: Pathological and MPO activity analyses revealed that IOP relieved LPS-induced uterine tissue injury. Quantitative reverse transcription-polymerase chain reaction was used to detect and quantitatively study the RNA information of mouse cells, which had high accuracy and sensitivity. From the test results, IOP does effectively control the release of pro-inflammatory cytokines such as interleukin-6 (IL-6), IL-1β, IL-8 and tumor necrosis factor-α (TNF-α), avoiding the body\'s immune response. Analysis of uterine tissue cell components also confirmed that the expression level of inflammatory mediator-induced nitric oxide synthase (iNOS) was also greatly reduced. Analysis of western blotting results of cell synthesis showed that IOP mainly inhibited the protein expression of TLR4 and myeloid differentiation factor 88 in the body.
CONCLUSIONS: This study proved that the mechanism of action of IOP is to inhibit the TLR4/NF-κB signaling pathway to reduce the release of pro-inflammatory cytokines from body cells, thereby alleviating the symptoms of endometritis induced by LPS. Thus, IOP may act as an effective drug in preventing and curing LPS-induced endometritis.