摘要
目的:探究白花丹素(plumbagi,PL)对博来霉素诱导的肺纤维化(pulmonary fibrosis,PF)的保护作用及其可能性机制。方法:将60只雄性C57BL/6小鼠随机分为:对照组(Control)、博来霉素组(bleomycin,BLM)、PL低剂量组(1 mg/kg)、PL高剂量组(2 mg/kg)。采用气管内注射BLM(3 mg/kg)复制小鼠PF模型,腹腔注射PL(1或2 mg/kg)3周,处死动物。HE与Masson染色观察肺组织形态学变化及胶原沉积情况。检测小鼠肺组织中超氧化物歧化酶(superoxide dis‐mutase,SOD)、谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)和羟脯氨酸(hydroxy‐proline,HYP)活性或含量。酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测小鼠肺组织中白介素-6(interleukin-6,IL-6)含量。免疫组化检测小鼠肺组织中核因子相关因子2(nuclear factor related factor 2,Nrf2)和NADPH氧化酶4(reduced nicotinamide adenine dinucleotide phosphate oxidase 4,NOX4)阳性细胞表达。Western blotting检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、I型胶原(collagen Ⅰ,Col Ⅰ)、Ⅲ型胶原(collagen Ⅲ,Col Ⅲ)、IL-6、转化生长因子-β_(1)(transforming growth factor-β_(1),TGF-β_(1))、p-Smad2、Nrf2和NOX4的蛋白表达。结果:与BLM组相比,PL治疗可减轻小鼠肺间质损伤及细胞外基质沉积,降低HYP含量(P<0.01,P<0.05),降低α-SMA、Col Ⅰ和Col Ⅲ的蛋白表达(P<0.01,P<0.05),减少IL-6的分泌(P<0.01),提高机体抗氧化能力(增强SOD和GSH的活性,减少MDA含量,P<0.01,P<0.05),显著下调TGF-β_(1)、p-Smad2和NOX4阳性细胞及蛋白表达(P<0.01,P<0.05),上调Nrf2阳性细胞及蛋白表达(P<0.01,P<0.05)。结论:PL可能通过调节TGF-β_(1)/Smad2及Nrf2/NOX4信号通路减轻炎症反应与胶原沉积,提高机体抗氧化能力,从而延缓PF进程。
AIM:To investigate the protective effect of plumbagin(PL)against pulmonary fibrosis(PF)and its possible mechanisms.METHODS:Sixty male C57BL/6 mice were randomly divided into control,bleomycin group(BLM),low dose PL group(1 mg/kg)and high dose PL group(2 mg/kg).The mice PF model was replicated using intratracheal injection of BLM(3 mg/kg),and then PL(1 or 2 mg/kg)was injected intraperitoneally for 3 weeks and the animals were executed.HE and Masson staining were used to observe morphological changes in lung tissue and collagen deposition.The activities or levels of superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA)and hydroxyproline(HYP)were measured in mouse lung tissue;ELISA for interleukin-6(IL-6)in mouse lung tissue.Immunohistochemical detection of nuclear factor-related factor 2(Nrf2)and reduced nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)positive cell expression in mouse lung tissue.The expression levels ofα-smooth muscle actin(α-SMA),collagen I(Col Ⅰ),collagen Ⅲ(Col Ⅲ),IL-6,transforming growth factor-β_(1)(TGF-β_(1)),p-Smad2,Nrf2 and NOX4 were detected by Western blotting.RESULTS:Compared with the BLM group,PL treatment attenuated lung parenchymal and interstitial injury and extracellular matrix deposition in mice,reduced HYP content(P<0.01,P<0.05),decreased protein expression ofα-SMA,collagen I and Ⅲ(P<0.01,P<0.05),diminished IL-6 secretion(P<0.01);improved the body's antioxidant capacity(increased SOD and GSH activity and decreased MDA content,P<0.01,P<0.05),significantly down-regulated TGF-β_(1),p-Smad2 and NOX4-positive cells and protein expression(P<0.01,P<0.05)and up-regulated Nrf2-positive cells and protein expression(P<0.01,P<0.05).CONCLUSION:PL may slow down the PF process by modulating the TGF-β_(1)/Smad2 and Nrf2/NOX4 pathways to attenuate inflammatory responses and collagen deposition and improve the body's antioxidant capacity.
作者
李慧
胡恒钊
俞婷婷
胡慧娴
王佳乐
吴晶
郝伟
LI Hui;HU Hengzhao;YU Tingting;HU Huixian;WANG Jiale;WU Jing;HAO Wei(Department of Functional Experimental Training Center,Basic Medical College,Wannan Medical College,Wuhu 241002,Anhui,China;School of Anesthesiology,Wannan Medical College,Wuhu 241002,Anhui,China;School of Medical Imaging,Wannan Medical College,Wuhu 241002,Anhui,China;School of pharmacy,Wannan Medical College,Wuhu 241002,Anhui,China)
出处
《中国临床药理学与治疗学》
北大核心
2025年第1期61-69,共9页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
2021年安徽省大学生创新创业训练计划项目(S202110368079)
2022年安徽省重点研究与开发计划项目(2022e07020036)。
