| Purpose:In this study,the rat model of chronic obstructive pulmonary disease was replicated by intratracheal instillation of lipopolysaccharide combined with cigarette smoking.Based on the fact that the lung inflammation runs through the whole process of occurrence and development of chronic obstructive pulmonary disease,we chose the clinical effective experience prescription-Hetiaoxuemaitongfei prescription of professor Li Qingsheng,a famous doctor in traditional Chinese medicine in Yunnan province,in which he treated chronic obstructive pulmonary disease by strengthening body resistance and nourishing blood to observe whether this prescription can block pulmonary vascular remodeling,regulate the airway mucosal immunity,repair intestinal mucosal barrier,and improve the inflammatory level of rats with chronic obstructive pulmonary disease.In order to preliminarily explain the mechanism of Hetiaoxuemaitongfei prescription by observing the relief level in lung inflammation.Methods:1.The rat model of chronic obstructive pulmonary disease was established by intratracheal instillation of lipopolysaccharide combined with cigarette smoking.The model was established for 56 days,and the success of the model was evaluated by observing the behavioral changes and pathological changes of lung tissue in rats.From the 57 th day,during the intervention period of traditional Chinese medicine,the cigarette smoking continued to be given every other day until the end of the experiment(that is,the 84 th day).2.Prepare the Hetiaoxuemaitongfei Decoction,and the intervention of traditional Chinese medicine was given by gavage once a day from the 57 th day.The control group and the model group were given the same dose of normal saline,and all groups gavage for 28 days.3.The blood cell analyzer was used to detect the count and percentage of neutrophils in rat blood.Serum IL-8,ET-1 and VEGF levels,SIgA levels in lung tissue and colon tissue were detected by ELISA.The expression levels of IL-1β,TNF-α and HIF-1α mRNA in rat lung tissue were detected by RT-PCR.The expression of Claudin-1 and ZO-1 in rat colon tissue was detected by immunohistochemistry.16 S rRNA gene high-throughput sequencing was used to detect the flora abundance of lung tissue and intestinal contents in rats.Results:1.Compared with the control group,the blood neutrophil count and neutrophil percentage in the model group rats increased significantly(P<0.01).Compared with the model group,the blood neutrophil count of rats in each dose group with traditional Chinese medicine intervention decreased(P>0.05),and the percentage of neutrophils decreased significantly(P<0.05);Compared with the control group,the serum IL-8 in the model group increased significantly(P<0.05).Compared with the model group,the IL-8 in each dose group with traditional Chinese medicine intervention decreased significantly(P<0.05).Compared with the control group,the relative expression of IL-1β and TNF-α mRNA in lung tissue of model group strikingly increased(P<0.01).Compared with model group,IL-1βm RNA in each dose group with traditional Chinese medicine intervention has a downward trend(P>0.05),especially obvious in low and middle dose groups(P<0.05),The relative expression of TNF-α mRNA decreased significantly in each dose group(P<0.01).Each dose group with traditional Chinese medicine intervention showed no obvious gradient change with dose change.2.Compared with the control group,the WT% and WA% of pulmonary arterioles stained by Victoria blue B + Van Gieson in the model group increased significantly(P<0.05),and compared with the model group,the WT% and WA% of each dose group intervened by traditional Chinese medicine has the downward trend(P>0.05),and the WT% of high dose group decreased obviously(P<0.05).Compared with the control group,the serum ET-1 of rats in the model group increased extremely(P<0.01)and VEGF increased extremely(P<0.05).Compared with the model group,ET-1 decreased in all dosage groups of TCM,especially in the low dosage group(P<0.01),and VEGF decreased extremely in all dosage groups(P<0.05).Compared with the control group,the relative expression level of HIF-1α mRNA in lung tissue of the model group increased significantly(P<0.05).Compared with the model group,the relative expression level of HIF-1α mRNA in each dose group decreased(P>0.05),especially obviously decreased in both the high and low dose groups(P<0.05).There was no gradient change with the dose in each dose group.3.Compared with the control group,SIgA of rat lung tissue in model group decreased significantly(P<0.05).Compared with the model group,SIg A of lung tissue in each dose group has trend to increase(P>0.05),especially in high dose group(P<0.05).As for the abundance of bacterial flora in rat lung tissue at phylum level,Proteobacteria and Firmicutes are dominant,and at the genus level,Ochrobactrum,Methylobacterium-Methylorubrum and Lactobacillus are the main species,followed by Ralstonia and Mycoplasma,among which,the relative abundance of 13 genera,such as Methylobacterium-Methylorubrum and Lactobacillus,decreased after modeling,and increased after intervention with traditional Chinese medicine,Ralstonia and Mycoplasma increased after modeling,and decreased after intervention with traditional Chinese medicine.4.Compared with the control group,the average optical densities of ZO-1 and Claudin-1 in the model group decreased strikingly(P<0.01).Compared with the model group,the average optical densities of ZO-1 and Claudin-1 in each dose group increased(P>0.05),especially obvious in the high dose group(P<0.01).Compared with the control group,SIgA of colon tissue in model group decreased significantly(P<0.01).compared with the model group,SIgA of colon tissue in each dose group has an upward trend(P>0.05),especially in high dose group(P<0.01).On the aspect of flora abundance of rat intestinal contents at phylum level,Firmicutes and Bacteroidetes are dominant.At the genus level,“Others”accounted for the majority of intestinal contents in rats(53.921%-69.053%),followed by unidentified-Ruminococcaceae and Turicibacter.Among them,the relative abundance of Methanosphaera and other two genera decreased after modeling,increased after intervention of Chinese medicine,increased after modeling of Blautia and Cloacibacillus,and decreased after intervention of traditional Chinese medicine.Conclusions:1.Hetiaoxuemaitongfei Prescription can improve lung inflammation and block pulmonary vascular remodeling in rats with chronic obstructive pulmonary disease.2.Hetiaoxuemaitongfei Prescription can slow down the level of chronic inflammation in the whole body in rats with chronic obstructive pulmonary disease by repairing intestinal mucosal barrier damage.3.The efficacy of Hetiaoxuemaitongfei Prescription in invigorating qi and nourishing blood,consolidating exterior and benefiting lung is reflected in blocking pulmonary vascular remodeling,regulating airway mucosal immunity and repairing intestinal mucosal barrier damage,and improving lung inflammation,thus delaying the progress of chronic obstructive pulmonary disease. |
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