| Objective: In this study,the main active ingredients of cordyceps drugs represented by Ophiocordyceps sinensis and Cordyceps militaris were selected: cordycepin and cordyceps polysaccharides.Through observation of rat serum and lung tissue inflammatory factors TNF-α,IL-6,IL-8 and CRP The expression of s Ig A in lung tissues and colon tissues,the expression of tight junction proteins ZO-1,Occludin and claudin-1 in colon tissues,the changes of gut microbiota,and a preliminary discussion on the regulation of cordycepin and cordyceps polysaccharide in COPD rats The biological basis of intestinal flora alleviating pulmonary inflammation provides a theoretical basis for clinical application of cordyceps drugs.Methods: 1.The rat model of COPD was replicated: Intratracheal infusion of lipopolysaccharide plus cigarette smoking was used.Rats except the control group were infused with lipopolysaccharide via tube on the 1st and 14 th day of modeling,and continued on days 2-13 and 15-56 Cigarette smoking,twice a day,half an hour each time.2.Drug intervention: After the completion of model building(day 57),the drug intervention group was given the low-dose(SL)and high-dose(SH)of cordycepin and the low-does(TL)and high-dose(TH)of cordyceps polysaccharide dissolved in normal saline by intragastric administration,and continued administration on 28 days,model group was given same amount of normal saline by gavage.3.Indicator detection:(1)Detect the pathological changes of rat lung tissue and colon tissue by HE staining method;(2)Calculate the thymus index and spleen index of rats by weighing;(3)Detect TNF-α,IL-6,IL-8 and CRP content in rat serum by ELISA method;(4)Detect the expression of TNF-α,IL-6,IL-8 and CRP in rat lung tissue by q PCR method;(5)Detect s Ig A content in rat lung tissue and colon tissue by ELISA method;(6)The expression of tight junction proteins Claudin-1,Occludinin and ZO-1,the colon tissue of rats was detected by WB method;(7)The abundance and diversity of the gut microbiota of rats were detected by 16 S r RNA gene high-throughput sequencing.Result: 1.Pathological changes of rat lung tissue and colon tissue 1.1 Pathological changes in lung tissue: Compared with control group,the alveolar interval in model group was widened or even broken,alveolar cavity expansion,alveolar fusion,and inflammatory cell infiltration in the blood vessels and peribronchial tissues of the lungs appeared.After administration intervention,the above-mentioned situation of the 4 treatment groups improved.1.2 Pathological changes of colon tissue: Compared with control group,intestinal mucosal inflammatory cells in model group were infiltrated,the submucosa was edema,and the local epithelium was broken off.After administration intervention,the above-mentioned situation of the 4 treatment groups improved.2.Changes of thymus and spleen index in rats The rat thymus index was lower in the model group than in the control group(P<0.01).After administration,compared with the model group,the SL group increased(P<0.05),the SH group increased(P<0.01),and the TL group increased(P <0.01),the TH group increased(P<0.01).The spleen index of rats in the model group was lower than that in the control group(P<0.05).After administration,compared with the model group,the SL group decreased(P>0.05),the SH group decreased(P>0.05),and the TL group decreased(P <0.05),the TH group decreased(P>0.05).3.Changes in the contents of TNF-α,IL-6,IL-8 and CRP in rat serum:The content of TNF-α in rat serum was higher in the model group than in the control group(P<0.01).After administration,compared with the model group,the SL group decreased(P>0.05),and the SH group decreased(P>0.05),the TL group decreased(P<0.01),and the TH group decreased(P<0.05).The content of IL-6 in rat serum was higher in the model group than in the control group(P<0.01).After administration,compared with the model group,the level of IL-6 in the SL group decreased(P<0.05),and the SH group decreased(P>0.05),decreased in the TL group(P<0.05),and decreased in the TH group(P<0.05).The content of IL-8 in rat serum was higher in the model group than in the control group(P<0.01).After administration,compared with the model group,the level of IL-8 in the SL group decreased(P<0.01),and the SH group decreased(P<0.01),the TL group decreased(P<0.01),and the TH group decreased(P<0.01).The content of CRP in rat serum was higher in the model group than in the control group(P<0.01).After administration,compared with the model group,the SL group decreased(P<0.01),and the SH group decreased(P<0.05).The TL group decreased(P<0.01),and the TH group decreased(P<0.01).4.Changes of TNF-α,IL-6,IL-8 and CRP m RNA expression in rat lung tissue Compared with the control group,the m RNA expression of TNF-α in rat lung tissues was increased in the model group(P<0.01).After administration,compared with the model group,the SL group decreased(P<0.01)and the SH group decreased(P<0.05),the TL group decreased(P<0.05),and the TH group decreased(P<0.05).Compared with control group,the m RNA expression of IL-6 in rat lung tissues was increased in the model group(P<0.05).After administration,compared with the model group,the SL group decreased(P<0.05)and the SH group decreased(P<0.05),the TL group decreased(P<0.05),and the TH group decreased(P<0.05).Compared with control group,m RNA expression of IL-8 in rat lung tissues was increased in the model group(P<0.05).After administration,compared with the model group,the SL group decreased(P<0.05)and the SH group decreased(P<0.05),the TL group decreased(P<0.05),and the TH group decreased(P<0.05).Compared with control group,m RNA expression of CRP in rat lung tissues was increased in the model group(P<0.01).After administration,compared with the model group,the SL group decreased(P<0.01)and the SH group decreased(P<0.01),the TL group decreased(P<0.01),and the TH group decreased(P<0.01).5.Changes of immunomolecule s Ig A content in mouse lung tissue and colon tissue The content of s Ig A in the lung tissue of rats was lower in the model group than in the control group(P<0.01).After administration,compared with the model group,the s Ig A content in the SL group was higher(P<0.05),and the SH group was higher(P<0.01),The TL group increased(P<0.01),and the TH group increased(P<0.01).The content of s Ig A in rat colon tissue was lower in the model group than in the control group(P<0.01).After administration,compared with the model group,the s Ig A content in the SL group was higher(P<0.01),and the SH group was higher(P<0.05),Increased in the TL group(P<0.01),and increased in the TH group(P<0.05).6.Relative expression levels of tight junction proteins ZO-1,Occludin and Claudin-1 in rat colon tissue Among the relative expression levels of tight junction proteins Claudin-1、Occludin and ZO-1 in the colon tissue of rats,the relative expression levels of the model group were lower than that of the control group.Compared with model group,relative expression levels of the four drug intervention groups all increased to varying degrees.7.Changes in the of rats:(1)Diversity changes: Compared with the control group,the gut microbiota diversity of rats in model group decreased;compared with model group,the gut microbiota diversity of rats in the four drug intervention groups increased.(2)Changes in abundance: From the perspective of changes in phylum level,compared with the blank group,the relative abundances of Firmicutes,Proteobacteria,and Verrucomicrobia in the model group decreased,while the relative abundances of Bacteroidetes and Euryarchaeota all increased.Compared with the model group,the relative abundances of Firmicutes and Actinobacteria in the four drug intervention groups all increased,and the relative abundances of Bacteroidetes,Euryarchaeota,Proteobacteria,and Verrucomicrobia all decreased.From the perspective of changes in genus level,compared with the control group,the relative abundances of Lactobacillus,Romboutsia,unidentified_Ruminococcaceae,Akkermansia,Turicibacter and Roseburia in the model group decreased,while the relative abundances of unidentified_Clostridiales,Methanobrevibacter,unidentified_Lachnospiraceae,unidentified_Prevotellaceae,Blautia and Bacteroides increased.Compared with the model group,the relative abundances of Lactobacillus,Methanobrevibacter,unidentified_Lachnospiraceae,Akkermansia,Blautia and Bacteroides all decreased,and the relative abundances of unidentified_Clostridiales,Romboutsia,unidentified_Ruminococcaceae,Turicibacter,Roseburia and unidentified_Prevotellaceae all increased.Conclusion: 1.Cordycepin and cordyceps polysaccharide can improve inflammation in COPD rats,and down-regulate the content of inflammatory factors TNF-α,IL-6,IL-8 and CRP.2.Cordycepin and cordyceps polysaccharide can regulate the immune function of COPD rats,increase thymus index,increase the content of s Ig A in lung tissue and colon tissue,and increase relative expression levels of tight junction proteins ZO-1,Occludin and Claudin-1 in colon tissue.3.Cordycepin and cordyceps polysaccharide can regulate the abundance and structure of gut microbiota in COPD rats. |
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