| Objective:In this study,we choose the Astragalus IV as the research drug to carry out animal experimental study,The general state,body weight,lung coefficient,lung tissue morphology and pathological changes were observed at different time points by comp arison,checking expression intensity of the Vascular endothelial cell marker(CD34),the Vascular endothelial growth factor and the Angiogenin-2,to investigate the role of Micr ovascular angiogenesis in the pathogenesis of IPF,to observe the effect of Astragalus I V on the expression of angiogenesis promoting factor and the mechanism of its effect on Microvascular angiogenesis,to research the function and mechanism of action on R at IPF that improved by astragaloside iv,To provide theoretical basis for further researc h on the treatment of idiopathic pulmonary fibrosis with Chinese herbal medicine.Method:After adaptive feeding 5d,21 rats were randomly selected as the blank gro up In 130 SPF rats,109 of the remaining rats were used to induce pulmonary fibrosis induced by modified tracheal puncture.105 rats were randomly divided into 5 groups:Model group,Hormone group,High,medium and low dose group of Astragalus IV,each g roup had 21 rats.The next day began to fill the stomach,Continuous gavage treatment7 d,14d,28 d,Rats were sacrificed at 7d,14 d,and28d,Collected and fixed specimens.IPF rat s were observed at different time points in general state and general form of lung tiss ue,weight and lung quality,the changes of body weight and lung coefficient were comp ared in rats.The pathological changes of Rat lung tissue were observed under microsco pe by HE and Masson staining,at the same time using the SABC method of immunoh istochemical technique and the combination of image analysis system,checking expressi on intensity of the Vascular endothelial cell marker(CD34),the Vascular endothelial gro wth factor and the Angiogenin-2.result:1.Observation of general state of ratsRats in the blank group were in good condition of diet and mental state,Sensitive reaction,the fur is Shiny,Smooth respiration.The rats were low-spirited on the second day,Slow reaction,activity decreased significantly,huddled up in this corner,dyspnea,groupget together.The spirit of model rats is still dispirited in the 7d, significantly reduced,piloerection, dry dull,shortness of breath,significantly reduced the amount of food,body weight loss,groups like get together,and the general condition of rats no turn for the b etter in 14 d,28d.The general state of Rats in 7d,14 d,28d,hormone group and low dose group of Astragalus group were better than homochronous model group.The overall sta te of the High and medium dose group were better than that of the model group, hor mone group and low dose group,and the Rats of High and medium dose group have good spirits,shiny fur,good diet,and smooth breathing,returned to normal in 28 d.2.Macroscopic observation of lung tissue in ratsRat lung tissues of blank group were pale pink, smooth surface, good elasticity, u niform texture, clear structure, no lesions in 7d,14 d,28d.In the 7d,the lung tissue of M odel group rats with marked hyperemia, edema, dark red color, the surface is rough, p atchy petechia, ecchymosis, a large number of uneven pale foci,the degree of lesion w as more serious than other groups;In the 14 d,the color of lung tissue of Model group rats is gray, the surface of the nodule like change, lung tissue is hard, poor flexibility,there were different real variable areas;In the 28 d,the lung tissue of Model group rats was significantly reduced volume, texture hard, uneven surface, nodular changes obvio usly, large ecchymosis.High, medium and low dose group of Astragalus and Hormone group were significantly better than Model group for the same period,at the same time,the high and middle dose group was better than the low dose group and the hormon e group,and the pathological changes of the low dose group were close to that in the same period.3.Comparison of body weight in ratsIn 7d, 14 d, 28 d,blank group rats body weight gradually increased(P<0.05), and s ignificantly higher than the same period of the model group, hormone group, middle a nd low dose group of Astragalus(P<0.01), but close to the High dose group of Astrag alus(P>0.05).In 7d, 14 d, 28 d,body weight of rats in model group were less than the s ame period in each treatment group(P<0.05), while the body weight of rats of High dose group of Astragalus were higher than the same period Hormone group and the low dose group of Astragalus(P<0.05),at the same time, the body weight of the rats in the dose group was larger than that of the same group and the low dose group(P<0.05),there was no significant difference between the low dose group and the hormo ne group(P>0.05).4.Comparison of lung coefficient in ratsIn 7d,14 d,28d,the lung coefficient of the blank group was less than other groups of the same period(P<0.05),and there was no significant change in lung coefficient at different time points(P>0.05);and the lung coefficient of model group was significa ntly higher than other groups of the same period(P<0.01);The High dose group of A stragalus was significantly less than other groups in the same period(P<0.01);The Rat lung coefficient of Middle dose group of astragaloside less than Low dose group of Astragalus and Hormone group of the corresponding period(P>0.05).The Rat lung coe fficient of Low dose group of astragaloside is close to the hameochronous model gro up(P>0.05).5.Results of HE staining and Masson staining were observedThere was no pathological changes in lung tissue at different time points in the b lank group.Model group:In 7d,Rat lung tissue damage and collapse, with a large numb er of exudate, alveolar septum widened significantly, a small amount of inflammatory cells, fibroblasts and fibroblasts aggregation;In 14 d,alveolar inflammation is lower than before,At the same time a large number of fibroblasts and fibroblasts proliferation,Ob vious pulmonary interstitial fibrosis;In 28 d,pulmonary interstitial fibrosis was more obvi ous.Compared with model group,At different time points, Alveolar inflammation and pu lmonary interstitial fibrosis of High, medium and low dose group of Astragalus and H ormone group were significantly decreased.Compared with hormone group,At different time points, Alveolar inflammation and pulmonary interstitial fibrosis of High and medi um dose group was significantly decreased.At different time points, the level of alveol ar inflammation and pulmonary interstitial fibrosis in low dose group was not compara ble to that in the same period.6.CD34 factor expression resultsIn 7d,14 d,28d,the expression intensity of CD34 in the lung tissue of each treatme nt group was significantly higher than that in the blank group(P<0.05),In model group, the expression intensity of CD34 in lung tissue was significantly higher than hameo chronous each medication group(P<0.01),the expression intensity of CD34 in the lung tissue of the high dose group and the middle dose group was lower than hameochron ous hormone group and the low dose group(P<0.05),and the expression of the high dose group was lowest(P<0.05),however,in the hormone group, the expression intensity of CD34 in the lung of rats was always the same as that of the low dose group(P>0.05).7.Expression of vascular endothelial growth factor(VEGF)In 7d,14 d,28d,the expression intensity of VEGF in the lung tissue of each treatme nt group was significantly higher than that in the blank group(P<0.05). In model gro up, the expression intensity of VEGF in lung tissue was significantly higher than hame ochronous Each medication group(P<0.01). The expression intensity of VEGF in the l ung tissue of the high dose group and the middle dose group was lower than that in the same period of the hormone group and the low dose group(P<0.05),and the expr ession of high dose group was lower than middle dose group(P<0.05).In the hormone group, the expression intensity of VEGF in the lung of rats was always the same as that of the low dose group(P>0.05).8.Expression results of the Angiogenin-2(Ang-2)Compared with the blank group, the cumulative light density of Ang-2 in the mo del group was significantly increased in 7d, 14 d and 28d(P<0.01).Compared with the model group, the expression of Ang-2 in the lung tissue of each drug group was sig nificantly decreased in 7d, 14 d and 28d(P<0.01).Compared with the hormone group, t he expression of Ang-2 in the lung tissue of the rats of the high and medium dose g roup was decreased(P<0.05),and the high dose group of Astragalus IV was weaker th an the middle dose group(P<0.05).the expression of Ang-2 of the low dose group was close to the hameochronous hormone group(P?0.05).Conclusion:1.IPF rat model was successfully induced by Injection of bleomycin with modifie d tracheal puncture method in this experiment.1.During the evolution of IPF in the experimental group,pulmonary alveolar inflammation in rats is most serious in 7d,alveolar inflammation gradually subsided in 14 d,pulmo nary interstitial fibrosis has appeared and developed gradually,pulmonary interstitial fibr osis is more obvious in 28 d.2.Modified tracheal puncture push injection of bleomycin induced pulmonary fibro sis in rats disease evolution process in the presence of pathological angiogenesis, path ological angiogenesis in 14 d is the most active,accompanied by pulmonary interstitial f ibrosis in rats of the whole process of pathological changes.3.In the experimental group, the incidence of IPF in rats,the sum of accumulated light density value of VEGF and the sum of Ang-2 accumulated light density were all enhanced in 7d, 14 d and 28 d,and 14 d enhance the most obvious,to promote the devel opment of pulmonary interstitial fibrosis in rats by promoting the VEGF and Ang-2 to promote the development of pulmonary interstitial fibrosis in rats,This is one of the mechanisms of the pathogenesis of IPF in rats.4.The effect of astragalus saponin can effectively inhibit the pathological angiogen esis in the lung tissue of IPF rats,so as to achieve the role of IPF in the treatment of rats,and its therapeutic effect is dose related. |
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