新橙皮苷对LPS诱导巨噬细胞炎症模型TLR4/NLRP3信号通路的影响  

Effects of neohesperidin on the TLR4/NLRP3 signaling pathway in LPS-induced macrophage inflammatory model

作　　者：黄超原 冯赟 雍秋红 杨希玲 杨喆彦萱 陆一慧 叶振昊 HUANG Chao-yuan;FENG Yun;YONG Qiu-hong;YANG Xi-ling;YANG Zhe-yan-xuan;LU Yi-hui;YE Zhen-hao(Department of Gastroenterology,The Second Affiliated Hospital of Guangzhou Universityof Chinese Medicine,Guangzhou 510120,Guangdong Province,China;The First Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong Province,China;Lingnan Medical Experimental Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong Province,China;Department of Gastroenterology,Shanghai General Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200080,China)

机构地区：[1]广州中医药大学第二附属医院脾胃病科,广东广州510120 [2]广州中医药大学第一临床医学院,广东广州510405 [3]广州中医药大学岭南医学实验研究中心,广东广州510405 [4]上海交通大学医学院附属第一人民医院消化内科,上海200080

出　　处：《中国临床药理学杂志》2024年第22期3275-3279,共5页The Chinese Journal of Clinical Pharmacology

基　　金：广东省科技创新战略专项基金重点资助项目(pdjh2023a0118)。

摘　　要：目的探讨新橙皮苷(NHP)对脂多糖(LPS)诱导的巨噬细胞炎症模型Toll样受体4(TLR4)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)信号通路的影响。方法将RAW264.7细胞分为对照组、模型组、低剂量实验组、高剂量实验组和MCC950组。对照组正常培养,其余组于培养基中加入100ng·mL^(-1)LPS以刺激获得巨噬细胞炎症模型。低、高剂量实验组分别用100、400μmol·L^(-1)的NHP干预;MCC950组用30μmol·L^(-1)的NLRP3抑制药(MCC950)干预。各组干预时间均为24h。干预结束后,用实时定量聚合酶链反应(qRT-PCR)实验法检测RAW264.7细胞中白细胞介素-1β(IL-1β)、TLR4和NLRP3mRNA的表达水平,用蛋白质印迹法检测RAW264.7细胞中TLR4、NLRP3蛋白的相对表达水平。结果对照组、模型组、低剂量实验组、高剂量实验组和MCC950组的IL-1βmRNA相对表达水平分别为1.00±0.08、6.45±1.19、3.87±0.55、1.96±0.32和3.26±0.16,TLR4mRNA相对表达水平分别为1.00±0.13、2.69±0.35、1.92±0.22、1.32±0.23和3.38±0.33,NLRP3mRNA相对表达水平分别为1.00±0.14、1.28±0.19、0.83±0.02、0.87±0.15和0.95±0.25,TLR4蛋白相对表达水平分别为0.63±0.05、0.86±0.04、0.68±0.08、0.64±0.08和0.71±0.08,NLRP3蛋白相对表达水平分别为0.44±0.02、0.66±0.03、0.56±0.07,0.52±0.05和0.54±0.07。模型组的上述指标与对照组比较,高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论NHP具有改善巨噬细胞炎症的作用,其机制与抑制TLR4/NLRP3信号通路有关。Objective To investigate the effect of neohesperidin(NHP)on the Toll-like receptor 4(TLR4)/NOD-like receptor family pyrin domain containing 3(NLRP3)signaling pathway in lipopolysaccharide(LPS)-induced macrophage inflammation model.Methods RAW264.7 cells were divided into five groups:Control group,model group,experimental-L group,experimental-H group and MCC950 group.The control group was cultured normally,while the other groups were stimulated with 100 ng·mL-1 of LPS to induce the macrophage inflammation model.The experimental-L,-H groups were treated with 100 and 400μmol·L^(-1) of NHP,respectively,while the MCC950 group was treated with 30μmol·L^(-1) of the NLRP3 inhibitor MCC950.All interventions lasted for 24 hours.After the intervention,quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression levels of interleukin-1β(IL-1β),TLR4 and NLRP3 in RAW264.7 cells.Western blotting was used to detect the protein expression levels of TLR4 and NLRP3 in RAW264.7 cells.Results The mRNA expression levels of IL-1βin the control,model,experimental-L,experimental-H and MCC950 groups were 1.00±0.08,6.45±1.19,3.87±0.55,1.96±0.32 and 3.26±0.16,respectively;the mRNA expression levels of TLR4 were 1.00±0.13,2.69±0.35,1.92±0.22,1.32±0.23 and 3.38±0.33,respectively;the mRNA expression levels of NLRP3 were 1.00±0.14,1.28±0.19,0.83±0.02,0.87±0.15 and 0.95±0.25,respectively;the protein expression levels of TLR4 were 0.63±0.05,0.86±0.04,0.68±0.08,0.64±0.08 and 0.71±0.08,respectively;the protein expression levels of NLRP3 were 0.44±0.02,0.66±0.03,0.56±0.07,0.52±0.05 and 0.54±0.07,respectively.The differences between the model group and the control group were statistically significant(all P<0.05);the differences between the experimental-H group and the model group were statistically significant(all P<0.05).Conclusion NHP can improve macrophage inflammation,and its mechanism is related to inhibition of TLR4/NLRP3 signaling pathway.

关 键 词：新橙皮苷 脂多糖 巨噬细胞 Toll样受体4 NOD样受体热蛋白结构域相关蛋白3 

分 类 号：R28[医药卫生—中药学]