2-吲哚啉酮衍生物对BLM诱导肺纤维化小鼠模型的治疗作用及机制研究

Therapeutic effect and Mechanism of 2-indolinone Derivatives on BLM-induced Pulmonary Fibrosis in Mice

【目的】探讨2-吲哚啉酮衍生物(PMID)对博莱霉素(BLM)诱导肺纤维化小鼠模型的治疗作用及机制。【方法】40只SPF级、3周龄SD雄性小鼠随机分为对照组、模型组及PMID低剂量组、PMID高剂量组,每组10只。模型组及PMID低、高剂量组小鼠采用BLM成功诱导建立肺纤维化模型后,PMID低、高剂量组小鼠给予5 mg/kg、20 mg/kg PMID溶液灌胃处理(2次/d,间隔6~8 h),连续28 d;对照组、模型组小鼠于同时间给予等量生理盐水灌胃处理。干预结束后,采用苏木精-伊红(HE)染色、胶原纤维(Masson)染色观察各组小鼠组织病理学变化及肺纤维化情况,并比较各组小鼠肺纤维化评分;采用实时荧光定量PCR法检测各组小鼠肺组织中Ⅲ型胶原(CollagenⅢ)mRNA水平;免疫印迹法检测各组肺组织中磷脂酰肌醇3-激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、缺氧诱导因子-1α(HIF-1α)蛋白水平。【结果】与对照组比较,模型组及PMID低、高剂量组小鼠肺纤维化评分、CollagenⅢmRNA水平及p-PI3K/PI3K、p-AKT/AKT、HIF-1α蛋白水平均升高(均P<0.05),且PMID低、高剂量组小鼠肺纤维化评分、CollagenⅢmRNA水平及p-PI3K/PI3K、p-AKT/AKT、HIF-1α蛋白水平低于模型组,且高剂量组上述指标水平低于低剂量组(均P<0.05)。【结论】PMID可改善BLM诱导的小鼠肺纤维化,可能与抑制PI3K/AKT/HIF-1α通路活化和胶原沉积有关。

【Objective】To explore the therapeutic effect and related mechanism of 2-indolinone derivative(PMID)on bleomycin(BLM)-induced pulmonary fibrosis in mice.【Methods】Forty SPF grade,3-week-old male SD mice were randomly divided into the control group,the model group and the PMID low and high dose groups,with 10 mice in each group.After the pulmonary fibrosis model was successfully induced by BLM in model group and PMID low and high dose groups,mice in PMID low and high dose groups were given 5 mg/kg and 20 mg/kg PMID solution by gavage(twice·d,with an interval of 6~8 h)for 28 days.While mice in the control group and the model group were given the same amount of normal saline at the same time.After drug intervention,the histopathological changes and pulmonary fibrosis of mice in each group were observed by hematoxylin eosin(HE)staining and collagen fiber(Masson)staining,then the pulmonary fibrosis scores of mice were compared.The mRNA level of collagenⅢin mouse lung was measured by real-time fluorescence quantitative PCR;Phosphatidylinositol 3-kinase(PI3K),p-PI3K,protein kinase B(AKT),p-AKT and hypoxia inducible factor-1α(HIF-1α)protein level were detected by Western blot.【Results】Compared to the control group,the pulmonary fibrosis score,CollagenⅢmRNA level,p-PI3K/PI3K,p-AKT/AKT and HIF-1αwere higher in the model group,low dose PMID group and high dose PMID groups(P<0.05).While the pulmonary fibrosis score,CollagenⅢmRNA level,p-PI3K/PI3K,p-AKT/AKT and HIF-1αof mice in low and high dose PMID groups were lower than those in the model group(P<0.05).In addition,the above index levels in the high-dose group were lower than those in the low-dose group(P<0.05).【Conclusion】PMID can improve BLM-induced pulmonary fibrosis in mice,which may be related to the inhibition of PI3K/AKT/HIF-1αpathway activation and inhibition of collagen deposition.