黄芪通过抗氧化抑制博莱霉素诱导的小鼠肺纤维化

摘要

AIM: To investigate the effect of Astragalus membranaceus on the balance of oxidation and antioxidation in bleomycin-induced pulmonary fibrosis in mice and the possible anti-fibrosis mechanism of Astragalus membranaceus.METHODS: Female KM mice (n=36) were randomly divided into 3 groups.The mice in control group were administered with saline aerosol intratracheally.The mice in fibrosis group were administered with bleomycin at dose of 3 mg/kg aerosol intratracheally.The mice in Astragalus membranaceus group were administered with bleomycin at dose of 3 mg/kg aerosol intratracheally and then intraperitoneal injected with Astragalus membranaceus parenteral solution at daily dose of 1.7 g/kg.All mice were sacrificed 14 d after the treatment, and the lungs and serum were collected for detection.Hematoxylin-eosin staining was performed in the lung tissue.The mRNA expression of superoxide dismutase 1/2/3 (SOD1/2/3), catalase (CAT), NADPH oxidase 2/4 (NOX2/4) and α-smooth muscle actin (α-SMA) was detected by RT-PCR, and the protein expression of α-SMA and NOX2/4 was determined by Western blot.The concentration of malondialdehyde (MDA) and total antioxidant capacity (T-AOC) in the serum were measured by a colorimetric method.RESULTS: The pathological injury was obviously observed in bleomycin group compared with control group, but was attenuated in Astragalus membranaceus group.The α-SMA mRNA and protein expression, MDA/T-AOC, NOX2, NOX4 and SOD3 mRNA expression, and NOX2 protein expression in bleomycin group were significantly higher than those in control group, while those in Astragalus membranaceus group were significantly lower than those in bleomycin group.The protein expression of NOX4 in bleomycin group was significantly lower than that in control group, while that in Astragalus membranaceus group was higher than that in bleomycin group.The mRNA expression of SOD1 and CAT in Astragalus membranaceus group and bleomycin group were decreased compared with control group.No significant difference of SOD2 mRNA expression among the 3 groups was observed.CONCLUSION: Astragalus membranaceus inhibits bleomycin-induced pulmonary fibrosis in mice by maintaining the balance of oxidation and antioxidation.%目的: 研究黄芪对博莱霉素诱导的肺纤维化小鼠氧化/抗氧化水平的影响,探讨黄芪抗纤维化的可能机制.方法: 将36只SPF级雌性昆明小鼠随机分为对照组(生理盐水气管内雾化)、博莱霉素组(博莱霉素3 mg/kg气管内雾化)和黄芪治疗组(博莱霉素3 mg/kg气管内雾化后黄芪注射液1.7 g·kg-1·d-1腹腔内注射),实验第14天收集小鼠肺组织及血清标本,取小鼠肺组织行HE和Masson染色;RT-PCR法测小鼠肺组织超氧化物歧化酶(SOD)1/2/3、过氧化氢酶(CAT)、NADPH氧化酶2/4(NOX2/4)和α-平滑肌肌动蛋白(α-SMA)的mRNA水平;Western blot法测α-SMA和NOX2/4蛋白表达水平;比色法检测血清丙二醛(MDA)和总抗氧化能力(T-AOC).结果: 博莱霉素组小鼠肺组织病理损伤较正常组明显加重,α-SMA mRNA和蛋白表达,MDA/T-AOC,NOX2、NOX4和SOD3 mRNA表达,以及NOX2蛋白表达较正常组显著上升,黄芪治疗组则显著逆转上述改变;博莱霉素组小鼠NOX4蛋白表达较正常组显著下降,而黄芪治疗组较博莱霉素组显著上升;博莱霉素组和黄芪治疗组小鼠SOD1和CAT mRNA表达均较正常组显著下降;SOD2 mRNA在3个组中表达的差异无统计学显著性.结论: 黄芪能够减缓博来霉素诱导的肺纤维化形成,其机制可能与调节氧化/抗氧化平衡有关.