摘要
本研究旨在验证西红花花瓣和花蕊提取物的抗炎效果,并探究其可能的作用机制.通过在二甲苯和脂多糖(LPS)诱导的小鼠炎症模型中评估提取物的治疗效果,连续给药14天,剂量为0.1和0.4g/kg.利用ELISA测定小鼠血清中TNF-α和IL-1β的浓度,并通过HE染色观察耳部和脾脏的病理变化.同时,通过Western blot检测NF-κB通路及NLRP3蛋白的表达变化.体外实验中,评估了西红花对LPS处理的RAW264.7细胞炎症模型的影响,使用CCK-8法进行细胞计数,并检测NLRP3、NF-κB及自噬相关蛋白的表达水平.研究结果显示,西红花提取物能有效治疗两种炎症模型,并通过下调NF-κB来减少NLRP3蛋白的表达,同时在体内外模型中降低自噬相关蛋白LC-3和beclin-1的表达.CCCP的加入减弱了西红花对NLRP3-NF-κB通路的影响.因此,本研究有助于阐明西红花提取物的抗炎作用机制.
Abstract
Objective:This study aimed to evaluate the anti-inflammatory effects of petal and stamen extracts of saffron crocus(Crocus sativus)and explore the underlying mechanism.Methods:Local and systemic inflammation models were used to investigate the anti-inflammatory effects of C.sativus.A xylene-induced inflammation model or lipopolysaccharide(LPS)-induced inflammation model was used in this study.C.sativus petal and stamen extracts were each administered to the mice in the xylene and LPS models by gavage for 14 d at 0.1 and 0.4g/kg doses,respectively.Enzyme-linked immunosorbent assay(ELISA)was used to measure the concentrations of tumor necrosis factor(TNF)-a and interleukin(IL)-1 β in mouse serum.Hematoxylin and eosin(H&E)staining was used to observe the pathological changes in the ear in the xylene-induced inflammation model and in the spleen in the LPS-induced inflammation model.NOD-like receptor thermal protein domain associated protein 3(NLRP3)protein levels within the nuclear factor-kappa B(NF-κB)pathway were assessed using western blotting.RAW264.7 cells were treated with LPS(5 μg/mL)and LPS+C.sativus(0.05,0.1,and 0.2mg/mL)for 24 h,and a Cell Counting Kit-8 was used to measure cell proliferation.Changes in NLRP3 and NF-κB levels were evaluated by western blotting.Results:Petal and stamen extracts of C.sativus attenuated the anti-inflammatory effects in local or systemic inflammatory models and repaired pathological changes in the ear in the xylene-induced inflammation model and spleen in the LPS-induced inflammation model.These extracts also decreased the concentrations of TNF-α and IL-1(3 in the mouse serum in the LPS-induced inflammation model.C.sativus downregulated NLRP3 protein level through the NF-κB pathway and downregulated LC-3 and BECLIN1 in vivo and in vitro.Carbonyl Cyanide3-ChloroPhenylhydrazone(CCCP)weakened the effects of C.sativus on the NLRP3-NF-κB pathway.Conclusion:C.sativus has anti-inflammatory effects and regulates the NLRP3-NF-κB pathway.
基金项目
National Natural Science Foundation of China(81873063)
Highlevel talents Research project of Hefei Normal University(2020rcjj30)
Key Project of Provincial Scientific Research Platform of Hefei Normal University in 2020(2020PTZD14)
Key Project of Universities Natural Science Foundation of Anhui province(KJ2021A0935)
Key Project of Universities Natural Science Foundation of Anhui province(KJ2021A0932)
Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-C-202009)
万方数据