丹芍化纤方对大鼠肺纤维化NF-κB与氧化应激的影响

Influences of Dan Shao Hua Xian Fang on nuclear factor-κB expression and oxidative stress in rats with pulmonary fibrosis

目的观察博莱霉素致大鼠肺纤维化中核转氯因子-κB(NF-κB)、超氧化物歧化酶(SOD)及丙二醛(MDA)的变化及丹芍化纤方对其的影响。方法选取SD大鼠30只,随机分为正常对照组(正常组)、肺纤维化模型组(模型组)和丹芍化纤方治疗组(丹芍组),每组各10只。正常组气管内滴注生理盐水,模型组和丹芍组气管内一次性灌注博来霉素5 mg/kg诱导肺纤维化,丹芍组于造模第2天开始每天给予丹芍化纤方灌胃(0.8 g/kg)治疗,正常组及模型组则给予等量生理盐水灌胃。实验第28天称大鼠体重,处死各组大鼠,称肺湿重并收集肺组织,行苏木精-伊红(HE)染色和马松(Masson)染色观察肺组织病理变化;比色法检测肺组织羟脯氨酸(HYP)、SOD及MDA变化;采用免疫组织化学的方法检测肺组织中NF-κB蛋白的表达情况。结果与正常组比较,模型组大鼠肺组织中MDA含量及NF-κB表达明显升高,而SOD酶活性则显著下降。与模型组比较,丹芍组大鼠肺组织中MDA含量及NF-κB蛋白表达明显降低,但仍较正常组高;SOD酶活性显著回升,但未恢复正常水平。结论丹芍化纤方早期应用可在一定程度上减轻博来霉素诱导的肺泡炎症反应及肺纤维化进展,对肺纤维化具有一定的治疗作用。丹芍化纤方对肺纤维化的干预机制可能与提高肺组织SOD酶活性、对抗氧化应激、抑制肺组织中NF-κB的表达、减轻肺泡炎症有关。

Objective To observe the changes of nuclear factor-KB （ NF-KB ）, superoxide dismutase （SOD） and malondialdehyde （MDA） in rats with bleomycin-induced pulmonary fibrosis, and the influences of Dan Shao Hua Xian Fang. Methods SD rats （ n = 30） were randomly divided into the normal group, model group and treatment group （each n = 10）. The normal group was intratracheally given instillation of normal saline solution, and model group and treatment group were intratracheally given once perfusion of bleomycin （5 mg/kg） for inducing pulmonary fibrosis. On the second.day of modeling, the treatment group was intragastrically given Dan Shao Hua Xian Fang （0. 8 g/kg）, and the normal group and model group were intragastrically given the same amount of normal saline solution. On the 28th day of the test, all rats were weighed and then killed. The wet weight of lung was weighed and lung tissue were collected. The pulmonary pathological changes were observed by using HE staining and Masson staining. The changes of hydroxyproline （HYP）, SOD and MDA were determined by using chromatoptometry, and the expression of NF-κB were detected by using immunohistochemistry assay.Results Compared with the normal group, the level of MDA and expression of NF-κB increased significantly and the activity of SOD decreased significantly in the model group. Compared with the model group, in the treatment group the level of MDA and expression of NF-κB decreased significantly but were still higher than those in the normal group. The activity of SOD rose again but had not reached normal level. Conclusion The early administration of Dan Shao Hua Xian Fang can alleviate to a certain extent the alveoli inflammatory reaction induced by bleomycin and pulmonary fibrosis development, so it has therapeutic effect on pulmonary fibrosis. The mechanism may be related to that Dan Shao Hua Xian Fang can improve the activity of SOD, antagonize oxidative stress, inhibit the expression of NF-κB and alleviate alveoli inflammatory.