肺淋巴循环障碍对矽肺发病进程的影响研究

Study on the influence of pulmonary lymphatic circulation disorder on the pathogenesis of silicosis

目的 探讨肺淋巴循环障碍对矽肺发病进程的影响。方法 采用胸导管结扎建立肺淋巴循环障碍模型。SPF级成年雄性SD大鼠24只随机分为对照组、结扎对照组、矽肺组、矽肺+结扎胸导管组(以下简称矽肺+结扎组)。每组各6只。对照组不做处理;结扎对照组给大鼠结扎胸导管后正常饲养;矽肺组大鼠采用动式染尘法建立矽肺模型(染尘仓内SiO_(2)粉尘浓度2000 mg/m^(3),每天给予大鼠动式染尘3 h);矽肺+结扎组大鼠先手术结扎大鼠胸导管,再进行动式染尘,染尘条件与矽肺组相同。HE染色观察肺组织病理变化;天狼猩红染色观察胶原沉积;免疫组化观察肺部淋巴管增生;Western blot检测大鼠肺组织中核转录因子(NF-κBp65)和血管内皮生长因子受体(VEGFR-3)、平滑肌肌动蛋白(α-SMA)的表达水平。结果 HE和天狼猩红染色结果显示,相较于对照组,矽肺组和矽肺+结扎组大鼠肺组织病变更为严重。免疫组化结果显示矽肺组和矽肺+结扎组大鼠肺组织内有淋巴管增生。Western blot结果显示肺组织中NF-κBp65和α-SMA的蛋白含量矽肺+结扎组均高于矽肺组,分别为(2.42±0.05)vs(1.80±0.01)、(2.81±0.06)vs(2.57±0.01)。相反,矽肺+结扎组大鼠肺组织内VEGFR-3的含量低于矽肺组(0.87±0.01)vs(0.97±0.03)。结论 结扎胸导管可使矽肺大鼠肺部出现淋巴循环障碍,加快矽肺发病进程。

Objective To explore the influence of pulmonary lymphatic circulation disorder on the pathogenesis of silicosis.Methods A pulmonary lymphatic circulation disorder model was established by thoracic duct ligation.Twenty-four SPF adult male SD rats were randomly divided into control group,ligation control group,silicosis groupand silicosis+ligation thoracic catheter group(hereinafter referred to as silicosis+ligation group)with 6 rats in each group.The control group was left untreated;the rats in the ligation control group were fed with thoracic duct ligation and the rats were fed normally;the rats in the silicosis group were established a silicosis model using dynamic dusting method(the concentration of SiO_(2) dust in the dusting chamber was 2000 mg/m^(3),and the rats were given dynamic dusting for 3 hours per day);the rats in the silicosis+ligation group were first surgically ligated to the rat thoracic duct,and then subjected to dynamic dust staining.The conditions for staining were the same as those in the silicosis group.HE staining was used to observe lung tissue pathological changes;Sirius scarlet staining was used to observe collagen Deposition;immunohistochemical observation of pulmonary lymphatic hyperplasia;Western blot was used to detect nuclear factor-κBp65(NF-κBp65),vascular endothelial growth factor receptor-3(VEGFR-3)andα-smooth muscle actin(α-SMA)in rat lung tissue.Results The results of HE and Sirius scarlet staining showed that the lung tissue lesions of the silicosis group and the silicosis+ligation group were more severe compared with the control group.The results of immunohistochemistry showed that there was lymphatic hyperplasia in the lung tissue of the rats in the silicosis group and the silicosis+ligation group.Western blot showed that the protein content of NF-κBp65 andα-SMA in lung tissues were higher in the silicosis+ligation group than that of the silicosis group(2.42±0.05)vs(1.80±0.01),(2.81±0.06)vs(2.57±0.01),respectively.On the contrary,the content of VEGFR-3 in the lung tissue of rats in the silicosis+ligation group was lower than that of the silicosis group(0.87±0.01)vs(0.97±0.03).Conclusion Ligation of the thoracic duct can cause lymphatic circulatory disorders in the lungs of silicosis rats and accelerate the onset of silicosis.