枇杷叶提取物干预博莱霉素诱导肺纤维化大鼠MH-S细胞炎症反应及机制研究

Effect of Loquat Leaf Extract on Inflammation of MH-S Cells Induced by Bleomycin in Rats with Pulmonary Fibrosis and Its Mechanism

本研究为探索枇杷叶提取物(EBJE)对肺纤维化(PF)大鼠巨噬细胞丝裂素活化蛋白激酶(MAPK)/核转录因子(NF-κB)信号通路和炎症损伤的调控作用。取Wistar大鼠120只,分为生理盐水(NS)组、模型(BLM)组、阳性对照组、EBJE高、中、低剂量组,每组20只。采用气管内一次性输注博来霉素(BLM,5 mg/kg)制备肺纤维化模型,造模成功后开始给药,1次/d,28 d后取材,分离肺泡巨噬(MH-S)细胞,Masson染色法检测各组肺纤维化病理进程;ELISA和RT-PCR检测各组肺泡巨噬细胞NF-κB、IL-17、TNF-α蛋白及mRNA表达情况;Western Blot检测MAPK/NF-κB信号通路相关蛋白p-p38 MAPK,p-38 MAPK,p-ERK1/2,ERK1/2表达情况;Person检验肺巨噬细胞MAPK蛋白表达水平。结果显示,与NS组相比,BLM组肺纤维化明显,肺部炎症细胞浸润比例增加,提示造模成功。与BLM组相比,EBJE高、中、低剂量组大鼠肺纤维化明显改善,炎症因子NF-κB、IL-17、TNF-α表达量降低,MAPK/NF-κB信号通路相关蛋白p-p38p-ERK1/2表达降低。EBJE可以缓解博莱霉素诱导大鼠肺纤维化导致的巨噬细胞炎症反应,同时调节巨噬细胞MAPK/NF-κB信号通路相关蛋白的表达,表明EBJE可能通过干预MAPK/NF-κB信号通路来缓解博莱霉素诱导的大鼠肺纤维化,其中EBJE高剂量组(3.81mL/kg)效果最优。

This study was conducted to observe the effects of loquat leaf extract(EBJE)on the regulation of macrophage mitogen-activated protein kinase(MAPK)/nuclear transcription factor(NF-κB)signaling pathway and inflammatory injury in rats with pulmonary fibrosis(PF).One hundred and twenty Wistar rats were divided into saline(NS)group,model(BLM)group,positive control group,and EBJE high,medium and low dose groups,with 20 rats in each group.The lung fibrosis model was prepared by intra-tracheal infusion of bleomycin(BLM,5 mg/kg),and the administration was started after successful moulding,once/d.The alveolar macrophage(MH-S)cells were isolated after 28 days,and the pathological process of lung fibrosis in each group was detected by Masson staining;the expression of TNF-αprotein and mRNA was detected by ELISA and RT-PCR,and the expression of MAPK/NF-κB signaling pathway-related proteins p-p38 MAPK,p-38 MAPK,p-ERK1/2,ERK1/2 was detected by Western Blot,and the expression level of MAPK protein in lung macrophages was examined by Person.Compared with the NS group,the BLM group showed significant lung fibrosis and an increased proportion of inflammatory cell infiltration in the lungs,suggesting successful modelling.Compared with the BLM group,lung fibrosis was significantly improved in the EBJE high,medium and low dose groups,with reduced expression of inflammatory factors NF-κB,IL-17 and TNF-α,and reduced expression of MAPK/NF-κB signalling pathway-related protein p-p38 p-ERK1/2.EBJE could alleviate the inflammatory response of macrophages caused by bleomycin-induced lung fibrosis in rats,while regulating macrophage.This suggests that EBJE may interfere with MAPK/NF-κB signaling pathway to alleviate bleomycin-induced pulmonary fibrosis in rats,with the best effect in the high dose group(3.81 mL/kg).