三叶青黄酮经p38MAPK和NF-κB途径抑制老年小鼠急性肺损伤

Inhibition effect of flavonoids from Radix tetrastigmae on acute lung injury of aged mice through p38MAPK and NF-κB pathway

目的探讨三叶青总黄酮(RTFs)对老年小鼠急性肺损伤(ALI)的保护作用及可能机制。方法采用老年C57BL/6J小鼠支气管滴注脂多糖(LPS)诱导ALI模型,三叶青黄酮灌胃给药3 d。取支气管肺泡灌洗液(BALF),瑞氏-吉姆萨染色计算白细胞数,ELISA检测炎症因子水平;ELISA和Western blot分析肺组织MAPKs、NF-κB蛋白表达,Trans AM检测核蛋白NF-κB活性。结果支气管滴注LPS成功诱导ALI模型,三叶青黄酮可明显减少BALF中白细胞和中性粒细胞数(P<0.01),降低IL-1β、IL-6、IL-12p40、TNF-α和s TNF-R1的分泌水平(P<0.01),改善肺组织病理损伤。三叶青黄酮明显抑制肺组织p38MAPK、NF-κB的磷酸化及NF-κB的DNA结合活性(P<0.01)。结论三叶青黄酮抑制p38MAPK、NF-κB炎症通路,保护LPS诱导的老年小鼠ALI。

Aim To investigate the protective effect of flavonoids from Radix tetrastigmae( RTFs) on lipopolysaccharide( LPS) induced acute lung injury( ALI)of aged mice and the mechanism. Methods Aged C57 BL /6J mice were bronchially instillated LPS to induce ALI. RTFs were orally administered to treat ALI.After 3 days,bronchoalveolar lavage fluid( BALF)was collected to enumerate leukocytes with Wright-Giemsa staining,and to detect inflammatory cytokines with ELISA. ELISA and Western blot methods were also used to detect the expression of MAPKs and NF-κB in lung tissues. The activity of NF-κB in nucleic protein extract was detected with Trans AM kit. Results ALI models were successfully induced through LPS instillation. RTFs significantly reduced leukocyte,especially neutrophil infiltration in BALF,inhibited IL-1β,IL-6,IL-12p40,TNF-α and s TNF-R1 secretion,and improved pathohistological change of lung tissues. Besides,RTFs significantly attenuated the phosphorylation of p38 MAPK,NF-κB and the activity of NF-κB.Conclusion RTFs inhibits LPS-induced ALI through p38 MAPK and NF-κB pathway and exhibits significant anti-inflammation effect on aged mice.