山柰酚减轻慢性阻塞性肺疾病模型小鼠的炎症反应

Kaempferol alleviates inflammatory response in chronic obstructive pulmonary disease model mice

目的:探讨山柰酚(kaempferol,Kae)对小鼠慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)的治疗作用及机制。方法:将雄性C57BL/6小鼠随机分为对照组、模型组和低、高剂量Kae治疗组,每组6只。对照组小鼠在正常条件下饲养;模型组小鼠暴露于香烟烟雾中12周;低、高剂量Kae治疗组小鼠在烟雾暴露前2 h分别予以25和100 mg/kg Kae灌胃。于第13周收集各组小鼠支气管肺泡灌洗液(bronchoalveolar lavage fluid,BLAF)和肺组织。HE染色观察肺组织病理学改变;ELISA试剂盒检测BLAF中促炎因子白细胞介素1β(interleukin-1β,IL-1β)、IL-5、单核细胞趋化蛋白1(monocyte chemoattractant protein-1,MCP-1)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)及肺组织中氧化应激标志物谷胱甘肽(glutathione,GSH)、一氧化氮(nitric oxide,NO)、丙二醛(malondialdehyde,MDA)和超氧化物歧化酶(superoxide dismutase,SOD)的水平;免疫荧光和Western blot分别检测肺组织中p-NF-κB p65蛋白的表达。结果:与对照组比较,模型组小鼠肺组织损伤评分显著升高(P<0.01),BLAF中MCP-1、TNF-α、IL-1β和IL-5水平及肺组织中MDA和NO水平、p-NF-κB p65的表达均显著增加(P<0.01),肺组织中SOD和GSH水平显著降低(P<0.01)。与模型组比较,低、高剂量Kae治疗组小鼠肺组织损伤评分显著降低(P<0.01),BLAF中MCP-1、TNF-α、IL-1β和IL-5水平及肺组织中MDA和NO水平、p-NF-κB p65表达均显著降低(P<0.05),肺组织中SOD和GSH水平显著增加(P<0.01),其中高剂量Kae治疗组上述指标改变尤为显著。结论:Kae对小鼠COPD能发挥一定的治疗作用,这一作用可能与其抗炎、抗氧化和抑制NF-κB信号通路相关。

AIM:To investigate the therapeutic effects of kaempferol(Kae)on chronic obstructive pulmonary disease(COPD)in mice and its mechanism.METHODS:Male C57BL/6 mice were randomly divided into 4 groups with 6 mice each:control group,model group,and low-and high-dose Kae treatment groups.The mice in control group were fed under normal conditions.The mice in model group were exposed to cigarette smoke for 12 weeks.The mice in low-and high-dose Kae treatment groups received intragastric administration of 25 and 100 mg/kg Kae 2 h before smoke exposure,respectively.At week 13,bronchoalveolar lavage fluid(BLAF)and lung tissue were collected.The pathological changes of the lung were observed by HE staining.The levels of pro-inflammatory cytokines interleakin-1β(IL-1β),IL-5,macrophage chemoattractant protein-1(MCP-1)and tumor necrosis factor-α(TNF-α)in BLAF,and oxidative stress markers glutathione(GSH),nitric oxide(NO),malondialdehyde(MDA)and superoxide dismutase(SOD)in lung tissue were detected by ELISA.The protein levels of p-NF-κB p65 in lung tissue were determined by immunofluorescence and Western blot.RESULTS:Compared with control group,the lung tissue injury score of the mice in model group was significantly increased(P<0.01),and the concentrations of MCP-1,TNF-α,IL-1β and IL-5 in BLAF,the contents of MDA and NO,and the protein level of p-NF-κB p65 in lung tissues were significantly increased(P<0.01).However,the activities of SOD and GSH in lung tissues were significantly decreased(P<0.01).Compared with model group,the lung tissue injury scores of the mice in low-and high-dose Kae groups were significantly decreased(P<0.01),and the concentrations of MCP-1,TNF-α,IL-1β and IL-5 in BLAF,the contents of MDA and NO,and the protein level of p-NF-κB p65 in lung tissues were significantly decreased(P<0.05),while the activities of SOD and GSH in lung tissues were significantly increased(P<0.01).CONCLUSION:Kaempferol exerts a therapeutic effect on COPD in mice,and this effect may be related to inhibiting inflammation,oxidative stress and NF-κB signaling pathway.