Effect Of Invigorating Qi And Activating Blood Circulation On PKD1/NF-κB/MnSOD Signal Pathway Induced By Bleomycin In IPF Rats

Objective: To observe the effect of Danggui Buxue Tang on lung histopathology and protein kinase D1(PKD1),nuclear transcription factor(NF-κB)and manganese superoxide dismutase(Mn SOD)mediated oxidative stress pathway in rats with Idiopathic pulmonary fibrosis induced by bleomycin,and to explore the mechanism of intervention of Idiopathic pulmonary fibrosis.Methods: Thirty-two male SPF SD rats were randomly divided into sham operation group,model group,Danggui Buxue Tang group and prednisone group,with 8 rats in each group.Except the sham operation group,the IPF rat model was established by intratracheal instillation of bleomycin in other groups.From the second day,Danggui Buxue Tang group was given Danggui Buxue Tang granule solution(0.81 g·kg·d),prednisone group(0.005 g·kg·d)was given prednisone solution(0.005 g·kg·d),sham operation group and model group were given the same volume of deionized water(1 m L·kg·d),for14 days.The general conditions of body weight and food intake of rats were recorded,and the syndrome scores of rats were scored according to the syndrome score scale of Qi deficiency and blood stasis syndrome.The fatigue state of rats was evaluated by weight-bearing swimming test,and the microcirculation perfusion of dorsal auricular vein was detected by speckle blood real-time imaging system(MBPU).After the detection of pulmonary function,blood was collected from the femoral artery and the serum was separated,and the lungs were obtained by thoracotomy.Weigh the lungs and calculate the lung index.The pathological changes of lung tissue were observed by hematoxylin-eosin staining(HE)and Masson staining,and the lung injury was evaluated by Szapiel score and Ashcroft score system,respectively.The content of malondialdehyde(MDA)in serum and the activities of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)in serum were measured,and the contents of hydroxyproline(HYP),hyaluronic acid(HA)and laminin(LN)in lung tissue were measured.The expressions of PKD1,NF-κB,Mn SOD m RNA and protein were detected by real-time fluorescence quantitative PCR(RT-q PCR)and Western blotting(WB).Results:1.General state of ratsThe coat color of rats in the sham operation group was bright,and the food intake and body weight increased steadily,while in the model group,the fur was not glossy,and the body weight and food intake decreased significantly after modeling,and increased gradually in the later stage.but it was significantly lower than that in the sham operation group(P < 0.01).2.State of Qi deficiency and blood stasis in ratsCompared with the sham operation group,the weight-bearing swimming time and MBPU in the model group,and the score of Qi deficiency and blood stasis scale increased in the model group(P < 0.01).Compared with the model group,the weight-bearing swimming time and MBPU in Danggui Buxue Tang group increased,and the score of Qi deficiency and blood stasis scale decreased(P < 0.01).Compared with prednisone group,the swimming time and MBPU of Danggui Buxue Tang group increased significantly,while the score of Qi deficiency and blood stasis syndrome decreased significantly(P <0.01).3.Comparison of pulmonary function of rats in each groupCompared with the sham operation group,Rrs,Ers,H and Rn in the model group increased significantly,while Crs and G decreased significantly(P < 0.01).Compared with the model group,Rrs,Ers,H and Rn in Danggui Buxue Tang group decreased significantly,while Crs and G increased significantly.4.Pathomorphological changes of lung tissue in rats of each groupCompared with the sham operation group,the Szapiel score,Ashcroft score and lung index of the model group increased,and the contents of HYP,HA and LN in the lung tissue increased significantly(P < 0.05 or P < 0.01).Compared with the model group,the Szapiel score,Ashcroft score and lung index in Danggui Buxue Tang group decreased significantly,and the contents of HYP,HA and LN in lung tissue decreased significantly(P < 0.05 or P <0.01).5.Changes of oxidative stress in rats in each groupCompared with the sham operation group,the content of serum MDA in the model group was significantly higher,and the activities of SOD,CAT and GSH-Px were significantly lower than those in the sham operation group(P <0.05 or P < 0.01).Compared with the model group,the content of serum MDA in Danggui Buxue Tang group decreased significantly,and the activities of SOD,CAT and GSH-Px increased(P < 0.05 or P < 0.01).6.Comparison of PKD1,NF-κB,Mn SODm RNA and protein expression in different groups of ratsCompared with the sham operation group,the expression levels of PKD1,NF-κB,Mn SOD m RNA and protein in the lung tissue of the model group were significantly higher than those of the sham operation group(P < 0.05 or P < 0 01).Compared with the model group,the expression levels of PKD1,NF-κB,Mn SOD m RNA and protein in lung tissue of Danggui Buxue Tang group decreased significantly(P < 0.05 or P < 0.01).Conclusion:1.The state of Qi deficiency and blood stasis is an important syndrome feature of IPF,and PKD1/NF-κB/Mn SOD signal pathway is an important regulatory mechanism of qi deficiency and blood stasis in IPF.2.The method of replenishing Qi and activating blood circulation can improve the degree of oxidative damage by regulating PKD1/NF-κB/Mn SOD pathway,thus delaying the pathological process of IPF.