鸡内金提取物通过调节肺泡巨噬细胞自噬水平减轻尘肺大鼠肺间质纤维化的机制

目的 探究鸡内金提取物通过调节肺泡巨噬细胞自噬水平减轻尘肺大鼠肺间质纤维化的机制.方法 选择30只雄性SD大鼠根据随机数字表法分为3组,每组10只.对照组为没有粉尘接触的健康SD大鼠;模型组采用一次性除尘方法建立大鼠矽肺模型;治疗组采用一次性除尘方法建立大鼠矽肺模型,每天灌服鸡内金提取物0.2 g.定量逆转录PCR(R...

Mechanism of Gallus gallus domesticus extract reducing pulmonary interstitial fibrosis in pneumoconiosis rats by regulating the autophagy level of alveolar macrophages

Objective To explore the mechanism of Gallus gallus domesticus extract reducing pulmonary interstitial fibrosis in pneumoconiosis rats by regulating the autophagy level of alveolar macrophages.Methods A total of 30 male Sprague-Dawley rats were selected and divided into 3 groups according to the random number table method,each group 10 rats.The control group was healthy SD rats without dust exposure;the model group established a rat silicosis model using a one-time dust removal method;the treatment group established a rat silicosis model using a one-time dust removal method,and was administered 0.2 g of Gallus gallus domesticus extract daily.The mRNA expression of autophagy-related genes(Beclin,LC3-Ⅱand LC3-Ⅰ)and apoptosis-related genes(Bcl-2)was analyzed by RT-qPCR;The viability of macrophages was detected by MTT at 6,12,24 h after culture.The scratch gap distance of different groups of cells was compared by scratch experiment at 12,24 h after culture;The protein expression levels of pro-inflammatory factors interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay.Western blotting was used to detect the expression of rat lung fibronectin,typeⅠcollagen,matrix metalloproteinase-9(MMP-9),matrix metalloproteinase-12(MMP-12),inducible nitric oxide synthase(iNOS)and Arginase-1(Arg1).Results Compared with the control group,Beclin,LC3-Ⅱ/LC3-ⅠmRNA expression,IL-1βand TNF-αlevels,and fibronectin,typeⅠcollagen,MMP-9,MMP-12 and iNOS protein expression levels in the model group were significantly higher,while Bcl-2 mRNA and Arg1 protein expression levels were significantly reduced(P<0.05);Compared with the model group,Beclin,LC3-Ⅱ/LC3-ⅠmRNA expression,IL-1βand TNF-αlevels,as well as fibronectin,typeⅠcollagen,MMP-9,MMP-12 protein and iNOS protein expression levels in the treatment group were significantly reduced,while the expression levels of Bcl-2 mRNA and Arg1 protein increased significantly(P<0.05).At 6,12,24 h after culture,the cell viability of the model group was lower than that of the control group(P<0.05),and the cell viability of the treatment group was higher than that of the model group(P<0.05).The distance between scratches in the model group at 12,24 h was lower than that of the control(P<0.05),and the distance between cell scratches in the treatment group was higher than that in the model group(P<0.05).Conclusion Gallus gallus domesticus extract exerts anti-inflammatory and anti-fibrotic effects by inducing autophagy in alveolar macrophages.Autophagy protects alveolar macrophages from silica-induced apoptosis by inhibiting the expression of fibronectin,typeⅠcollagen,MMP-9 and MMP-12,thereby further reducing the secretion of inflammatory factors caused by silica particles And final fibrosis.