Abstract：
BACKGROUND: Millingtonia hortensis L.f., commonly known as tree jasmine or Indian cork tree, is native to South Asia and Southeast Asia. Traditionally, its stem bark, leaves, and roots are employed for pulmonary, gastrointestinal, and antimicrobial purposes, while the flowers are used in treating asthma and sinusitis.
OBJECTIVE: The underlying anti-inflammatory mechanisms of M. hortensis remain relatively unexplored. Therefore, we studied the anti-inflammatory effects of M. hortensis and the molecular mechanisms of its ethanol extracts (Mh-EE) both in vitro and in vivo.
METHODS: Nitric oxide (NO) production was assessed using Griess reagent, while cell viability of RAW264.7 cell and HEK293T cells were determined via the MTT assay. Constituent analysis of Mh-EE using GC/MS-MS and HPLC, and mRNA expression of inflammatory cytokines was measured through PCR and RT-PCR. Protein levels were analyzed using western blotting. The thermal stability of Mh-EE was evaluated by CESTA. Lastly, a gastritis in vivo model was induced by HCl/EtOH, and protein expression levels were measured using western blotting.
RESULTS: Mh-EE significantly reduced NO production in LPS-induced RAW264.7 cells without substantially affecting cell viability. Additionally, Mh-EE decreased the expression of proinflammatory factors, such as iNOS, IL-1β and COX2. Furthermore, Mh-EE downregulated TLR4 expression, altered MyD88 recruitment, and suppressed phosphorylation of Syk, IKKα, IκBα and AKT. Simultaneously, Mh-EE also attenuated NF-κB signaling in HCl/EtOH-induced mice.
CONCLUSIONS: Mh-EE exerts anti-inflammatory effects by suppressing p-Syk in the NF-κB pathway, and it has potential as a novel treatment agent for inflammatory diseases.
OBJECTIVE: The underlying anti-inflammatory mechanisms of M. hortensis remain relatively unexplored. Therefore, we studied the anti-inflammatory effects of M. hortensis and the molecular mechanisms of its ethanol extracts (Mh-EE) both in vitro and in vivo.
METHODS: Nitric oxide (NO) production was assessed using Griess reagent, while cell viability of RAW264.7 cell and HEK293T cells were determined via the MTT assay. Constituent analysis of Mh-EE using GC/MS-MS and HPLC, and mRNA expression of inflammatory cytokines was measured through PCR and RT-PCR. Protein levels were analyzed using western blotting. The thermal stability of Mh-EE was evaluated by CESTA. Lastly, a gastritis in vivo model was induced by HCl/EtOH, and protein expression levels were measured using western blotting.
RESULTS: Mh-EE significantly reduced NO production in LPS-induced RAW264.7 cells without substantially affecting cell viability. Additionally, Mh-EE decreased the expression of proinflammatory factors, such as iNOS, IL-1β and COX2. Furthermore, Mh-EE downregulated TLR4 expression, altered MyD88 recruitment, and suppressed phosphorylation of Syk, IKKα, IκBα and AKT. Simultaneously, Mh-EE also attenuated NF-κB signaling in HCl/EtOH-induced mice.
CONCLUSIONS: Mh-EE exerts anti-inflammatory effects by suppressing p-Syk in the NF-κB pathway, and it has potential as a novel treatment agent for inflammatory diseases.
摘要：
背景：MillingtoniahortensisL.f.，通常被称为茉莉树或印度软木树，原产于南亚和东南亚。传统上,它的茎皮，叶子,根用于肺部，胃肠,和抗菌目的，而花用于治疗哮喘和鼻窦炎。
目的：关于木耳分枝杆菌的抗炎机制还未被研究。因此,我们在体外和体内研究了M.hortensis的抗炎作用及其乙醇提取物（Mh-EE）的分子机制。
方法：使用Griess试剂评估一氧化氮（NO）的产生，同时通过MTT法测定RAW264.7细胞和HEK293T细胞的细胞活力。使用GC/MS-MS和HPLC分析Mh-EE的成分，通过PCR和RT-PCR检测炎性细胞因子的mRNA表达。使用蛋白质印迹分析蛋白质水平。通过CESTA评价Mh-EE的热稳定性。最后,由HCl/EtOH诱导的胃炎体内模型，使用蛋白质印迹法测量蛋白质表达水平。
结果:Mh-EE显著降低LPS诱导的RAW264.7细胞中的NO产生,而基本上不影响细胞活力。此外,Mh-EE降低促炎因子的表达,例如iNOS，IL-1β和COX2。此外，Mh-EE下调TLR4表达，改变了MyD88的招募，抑制Syk的磷酸化，IKKα,IκBα和AKT。同时,Mh-EE还减弱HCl/EtOH诱导的小鼠中的NF-κB信号传导。
结论：Mh-EE通过抑制NF-κB通路中的p-Syk发挥抗炎作用，它具有作为炎症性疾病的新型治疗剂的潜力。
目的：关于木耳分枝杆菌的抗炎机制还未被研究。因此,我们在体外和体内研究了M.hortensis的抗炎作用及其乙醇提取物（Mh-EE）的分子机制。
方法：使用Griess试剂评估一氧化氮（NO）的产生，同时通过MTT法测定RAW264.7细胞和HEK293T细胞的细胞活力。使用GC/MS-MS和HPLC分析Mh-EE的成分，通过PCR和RT-PCR检测炎性细胞因子的mRNA表达。使用蛋白质印迹分析蛋白质水平。通过CESTA评价Mh-EE的热稳定性。最后,由HCl/EtOH诱导的胃炎体内模型，使用蛋白质印迹法测量蛋白质表达水平。
结果:Mh-EE显著降低LPS诱导的RAW264.7细胞中的NO产生,而基本上不影响细胞活力。此外,Mh-EE降低促炎因子的表达,例如iNOS，IL-1β和COX2。此外，Mh-EE下调TLR4表达，改变了MyD88的招募，抑制Syk的磷酸化，IKKα,IκBα和AKT。同时,Mh-EE还减弱HCl/EtOH诱导的小鼠中的NF-κB信号传导。
结论：Mh-EE通过抑制NF-κB通路中的p-Syk发挥抗炎作用，它具有作为炎症性疾病的新型治疗剂的潜力。
