黄连素激活Nrf2通路缓解新生大鼠哮喘模型氧化应激反应和对心肺组织的保护作用

Berberine activates Nrf2 pathway to relieve oxidative stress response and protect heart and lung tissues in the newborn asthma rat model

目的:探讨黄连素(berberine,BB)对新生大鼠哮喘模型氧化应激反应的影响和对心肺组织的保护作用以及潜在的分子机制。方法:将新生SD雄性大鼠随机分成健康对照组、健康加药组、哮喘模型组和模型加药组;卵清蛋白诱导新生大鼠哮喘模型;收集肺泡灌洗液(broncho alveokar lavage fluid,BALF)并分析细胞类型和细胞数目;苏木伊红(hematoxylin eosin,HE)染色观察心肌组织和肺组织病理变化;末端标记法(terminal deoxynucleoitidyl transferase mediated nick end labeling,TUNEL)染色检测心肌组织和肺组织细胞凋亡情况;试剂盒检测血清中超氧化物歧化酶(superoxide dismutase,SOD)的活性和丙二醛(malondialdehyde,MDA)、一氧化氮(nitric oxide,NO)的含量;蛋白质印记检测肺组织中Kelch样ECH相关蛋白1(Kelch-like ECH-associated protein 1,Keap1)、NFE2相关因子2(NF-E2 related factor 2,Nrf2)和血红素氧合酶1(heme oxygenase 1,HMOX-1)的表达水平。结果:哮喘模型组与对照组相比,肺泡灌洗液中炎性细胞的数目显著增加;心肌组织细胞排列不规则,部分细胞结构不清晰;肺组织中有大量的炎性细胞浸润,黏膜下水肿,气道上皮断裂脱落,支气管壁明显增厚;心肌组织和肺组织中凋亡细胞比例显著增加;血清中SOD的活性显著降低,MDA和NO的含量显著增加;肺组织中Keap1、Nrf2和HMOX-1的表达水平显著升高。模型加药组与模型组相比,肺泡灌洗液中炎性细胞的数目明显减少;心肌组织较规则,细胞排列较整齐;肺组织中仍有部分炎性细胞浸润,部分肺泡壁增厚;心肌组织和肺组织中凋亡细胞比例显著降低;SOD的活性显著升高,MDA和NO的含量显著降低;Keap1、Nrf2和HMOX-1的表达水平显著升高。结论:黄连素可激活Nrf2通路,缓解新生大鼠哮喘模型的氧化应激反应,对心肺组织具有保护作用。

OBJECTIVE To investigate the potential cardiopulmonary protective effects of berberine on oxidative stress in the asthmatic neonatal rat model,and to explore the potential molecular mechanism.METHODS Male neonatal SD rats were randomly divided into the healthy control group,healthy medication group,asthma model group and model medication group.Ovalbumin was used to induce the newborn asthma rat model;Broncho alveolar lavage fluid(BALF)was collected,and cell type and number were analyzed.Pathological changes in the myocardial tissues and lung tissues were observed through hematoxylin eosin staining.Apoptosis of cardiomyocytes and lung tissues was detected with TUNEL staining;superoxide dismutase(SOD)activity and contents of malondialdehyde(MDA)and nitric oxide(NO)in the serum were detected with related kits.The expression levels of Kelch-like ECH-associated protein 1(Keap1),NFE2 related factor 2(Nrf2)and heme oxygenase 1(HMOX-1)in lung tissues were detected with Western blot.RESULTS Compared with the control group,the number of inflammatory cells in the alveolar lavage fluid increased significantly in the asthma model group.The myocardial tissue cells showed irregular arrangement and some cell structures were not clear.Infiltration of a large number of inflammatory cells was observed in lung tissues,with submucosal edema,airway epithelium rupture and shedding,and significantly thickened bronchial wall.The ratio of apoptotic cells in lung tissues increased significantly.The activity of SOD in the serum decreased significantly,and the content of MDA and NO increased significantly.And the expression levels of Keap1,Nrf2 and HMOX-1 in lung tissues increased significantly.Compared with the model group,the number of inflammatory cells in the alveolar lavage fluid in the model group was significantly reduced;the myocardium was more regular and the cells were arranged more regularly;some inflammatory cell infiltration in the lung tissue and some alveolar wall thickening still existed.The apoptotic cell ratio in the myocardium and lung tissues was significantly decreased.The activity of SOD was significantly increased,and the concentrations of MDA and NO were significantly reduced.The expression levels of Keap1,Nrf2 and HMOX-1 increased significantly.CONCLUSION Berberine can activate the Nrf2 pathway and alleviate oxidative stress in the neonatal asthma rat model,so as to protect cardiopulmonary tissues.