参莲方对颗粒物暴露致大鼠急性肺损伤的保护作用

Protective Effect of Shenlian Prescription on Acute Lung Injury Induced by Particulate Matter Exposure in Rats

目的:观察参莲方对颗粒物(PM)暴露致大鼠急性肺损伤的保护作用并初步探究其保护机制。方法:雄性SD大鼠50只,随机分为正常组、模型组、参莲方低、高剂量组(4.32、8.64 g·kg^(-1))、罗氟斯特组(3.46 mg·kg^(-1)),每组10只。预给药灌胃1周后,于第8天和第11天除正常组滴注生理盐水外,其余组气管滴注PM混悬液建立PM暴露致大鼠急性肺损伤模型。造模后继续给药至实验结束。末次染毒48 h后检测大鼠肺功能。随后处死大鼠,观察肺部形态学变化,苏木素-伊红(HE)染色检测肺组织病理变化;免疫组化观察肺部CD68表达;酶联免疫吸附测定法(ELISA)检测血清肺损伤标志物肺表面活性物质A(SP-A)、克拉氏细胞分泌蛋白16(CC16)水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织炎症因子白细胞介素-1α(IL-1α)、IL-6、IL-18和单核细胞趋化蛋白-1(MCP-1)mRNA表达。结果:与正常组比较,模型组大鼠肺功能下降,肺组织出现明显组织结构损伤,并有明显的PM沉积和CD68阳性细胞浸润,肺组织炎症因子IL-1α、IL-6、IL-18、MCP-1 mRNA表达显著升高(P<0.01);与模型组比较,参莲方各给药组大鼠肺功能指标改善(P<0.05,P<0.01),肺组织形态学改善,PM沉积减少,组织病理结构改善,CD68阳性细胞浸润未显著减少,炎症因子IL-1α、IL-6、IL-18、MCP-1 mRNA水平显著降低(P<0.01)。结论:参莲方可保护PM暴露造成的大鼠肺损伤,改善组织形态,减少肺部PM沉积,减少炎症因子表达。

Objective:To observe the protective effect of Shenlian prescription on acute lung injury induced by particulate matter(PM)exposure in rats and explore the mechanism.Method:Fifty male SD rats were randomly divided into the control group,model group,Shenlian low-dose group(4.32 g·kg^(-1)),Shenlian high-dose group(8.64 g·kg^(-1)),and roflumilast group(3.46 mg·kg^(-1)),with 10 in each group.Pre-administration with drugs by gavage was performed for one week.On the 8thand 11thdays,the control group was instilled with normal saline in the trachea and the other groups with PM suspension to establish a rat model of acute lung injury induced by PM exposure.After modeling,drugs were given continuously until the end of the experiment.Fortyeight hours after the last exposure,the lung function of rats was detected.Then the rats were sacrificed and the lung morphological changes and pathological changes by hematoxylin-eosin(HE)staining were observed.CD68expression in lung was detected by immunohistochemistry,and the levels of lung injury markers surfactant protein A(SP-A)and Clara cell protein16(CC16)in serum were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression of interleukin-1α(IL-1α),IL-6,IL-18,and monocyte chemoattractant protein-1(MCP-1)in lung tissue was measured by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:Compared with those in the control group,the rats in the model group had decreased lung function and obvious structural damage of lung tissue,PM deposition,and infiltration of CD68positive cells.The expressions of IL-1α,IL-6,IL-18,and MCP-1 in lung tissue were increased(P<0.01).Compared with the model group,Shenlian prescription low and high doses restored the rats’lung function injury(P<0.05,P<0.01),improved lung morphological and pathological structure,and reduced PM deposition.Infiltration of CD68 positive cells in lung was not significantly decreased.The levels of inflammatory factors IL-1α,IL-6,IL-18,and MCP-1 in lung were lowered(P<0.01).Conclusion:Shenlian prescription could protect the rats’lung injury caused by PM exposure,improve lung morphology,and reduce PM deposition and inflammatory factor expression.