Effect Of Method Of Reinforcing Earth To Generate Matal On Ghrelin-Obestatin Signal Pathway Of COPD Rat With Deficiency Of Both Lung And Spleen Based On The Brain-Gut Axis

Objective:To duplicate the model of chronic obstructive pulmonary disease(COPD)deficiency in lung and spleen,observe the effects of method of reinforcing earth to generate matal on lung function,pathological morphology of lung,hypothalamus and duodenum in model rats,to further study the expression level of genes and proteins related to Ghrelin-Obestatin system in the brain-intestine axis of model rat induced by method of reinforcing earth to generate matal,expected to discuss the mechanism of the prevention and treatment of COPD with malnutrition by the method of method of reinforcing earth to generate matal from the perspective of brain-intestine interaction.Methods:75 SPF Wistar rats were randomly divided into blank control group,lung and spleen deficiency observation group and drug intervention group(divided into 28 d,35d and 42 d groups according to different drug intervention time),each group of 18.COPD rats of both spleen and spleen deficiency type were duplicated by way of combination of disease and syndrome.Rats of blank control group were given inhaled air and intratracheal instillation of equal volume of normal saline and equal volume of normal saline,the observation group and drug intervention group were smoked + Intratracheal instillation of lipopolysaccharide + perfusion of cold senna extract to establish lung and spleen deficiency COPD rat model;drug intervention group given Shenling Baizhu power,the rats were sacrificed on the 28 th day,35 th day and 42 nd day after drug intervention respectively.The rats were sacrificed,the hypothalamus and duodenum of rats were taken,the expression of Ghrelin,Obestatin and its receptors(GHSR,GPR39)mRNA and protein in rat hypothalamus and duodenum were detected by RT-PCR,immunohistochemistry and immunoblotting.Results:1.Changes in rat lung function(awake state): Compared with the blank control group,the pulmonary function TV,EV,F,RT,PIF,PEF in the observation group with lung and spleen deficiency were significantly higher(P<0.05),the difference was statistically significant,and the EF50 and AV were significantly decreased(P<0.05);Compared with the observation group with lung and spleen deficiency,Shenqi Baizhu power 28 d group had higher RT(P<0.05),TV,EV,F,PIF,and PEF were significantly lower(P<0.05).The AV and RT of Shenqi Baizhu power 35 d group increased(P<0.05),EF50,TV,EV,F,PIF,and PEF decreased(P<0.05).The AV of Shenqibaizhu power 42 d group increased(P<0.05),EF50,F,RT,PIF,PEF decreased(P<0.05),and the difference was statistically significant.2.Rat lung tissue,hypothalamus,duodenum histopathological changes:2.1Pathological changes of lung tissue in rats:The morphology of bronchus and alveoli in the blank control group was normal,the structure of bronchus and alveoli were normal,the epithelial structure of bronchial mucosa was intact,the cilia arranged neatly,no gland hyperplasia,alveolar size,normal structure,no inflammatory cell infiltration;lung and spleen deficiency group showed bronchial mucosal epithelial disorders,shedding,ciliary lodging,goblet cells increased,gland hypertrophy,a large number of inflammatory cell infiltration,smooth muscle hypertrophy,Alveolar wall thinning,rupture,alveolar swelling,multiple alveolar spaces increase,mutual integration,the formation of bulla;Shenling Baizhu power 28 d group showed bronchial mucosal epithelial disorder,shedding,still inflammatory cell infiltration,rupture of the alveolar wall compared with the observation group of lung and spleen deficiency;Shenling Baizhu power 35 d group showed bronchial mucosal epithelial shedding,inflammatory cell infiltration,alveolar wall thinning,alveolar enlargement,part of alveolar fusion,the degree of the model group lighter;Shenling Baizhu power 42 d group showed partial bronchial epithelial shedding,a small amount of inflammatory cell infiltration,bulla degree of lung lighter than lung and spleen deficiency group.2.2Pathological changes of hypothalamus in rats:Hypothalamic neuronal cells in the blank control group are structurally intact and neatly arranged,all kinds of cells are clearly visible and evenly distributed;hypothalamus changes in rats of both lung and spleen deficiency group and drug intervention group did not change much.2.3Rat duodenum pathological changes:The morphology and structure of the duodenum mucosa epithelium in the blank control group were intact and normal,the villi had a clear structure,no inflammatory cell infiltration;lung and spleen deficiency group showed duodenal mucosal damage,epithelial cells degeneration and necrosism,interstitial inflammatory cells increase,mild edemam,submucosa and muscular structure integrity;Shenling Baizhu power 28 d group showed duodenal villi bending,villous surface epithelial deletion,interstitial inflammatory cell infiltration,the degree lighter than lung and spleen deficiency observation group;Shenling Baizhu power 35 d group shows part of duodenal mucosal structure damage,interstitial inflammatory cell infiltration;Shenling Baizhu power 42 d group visible occasional rupture of villi,epithelial cells shed less than lung and spleen deficiency observation group,a small amount of inflammatory cell infiltration.3.Expression of Ghrelin,Obestatin,GHSR and GPR39 mRNA in hypothalamus and duodenum in rats:3.1 Expression of Ghrelin,Obestatin,GHSR and GPR39 mRNA in rat hypothalamus:Compared with the blank control group,the expression levels of Ghrelin,GHSR and GPR39 mRNA in the hypothalamus of the observation group were significantly decreased(P<0.05);the levels of Ghrelin,GHSR and GPR39 m RNA in the hypothalamus of each intervention group were significantly higher than those of the observation group(P<0.05),and there was significant difference between the two groups(P<0.05).3.2 Expression of Ghrelin,Obestatin,GHSR and GPR39 mRNA in rat duodenum:Compared with the blank control group,the expressions of Ghrelin,GHSR and GPR39 mRNA in the duodenum of the lung-spleen deficiency group were decreased(P<0.05);the levels of Ghrelin,GHSR and GPR39 mRNA in each intervention group were significantly higher than those in the observation group(P<0.05),and the contents of Ghrelin,GHSR and GPR39 mRNA increased significantly with the intervention time increasing(P<0.05).4.Mean optical density expression of Ghrelin,Obestatin,GHSR and GPR39 in rat hypothalamus and duodenum:4.1Expression of Ghrelin,Obestatin,GHSR and GPR39 in rat hypothalamus:Compared with the blank control group,the mean optical density of Ghrelin,GHSR and GPR39 in the hypothalamus of the observation group was significantly decreased(P<0.05),and the average optical density of Obestatin increased(P<0.05);the average optical density of Ghrelin,GHSR and GPR39 in hypothalamus increased(P<0.05),and the average optical density of Obestatin decreased compared with the observation group(P<0.05),the difference between each index group was significant(P<0.05).4.2Expression of Gdrelin,Obestatin,GHSR and GPR39 in rat duodenum:Compared with the blank control group,the average optical density of Ghrelin,GHSR,Obestatin,and GPR39 in the duodenal tissue of the observation group of lung and spleen deficiency was significantly lower(P<0.05);compared with the observation group of lung and spleen deficiency,There was no significant difference in Ghrelin,GHSR,GPR39 between duodenum of the Shenling Baizhu power 28 d group(P>0.05).Ghrelin and GHSR were not statistically significant(P>0.05)in the Shenling baishu power35 d group(P>0.05).The average optical density of Ghrelin,GHSR,Obestatin,and GPR39 increased in 42 d group(P<0.05),and the difference was statistically significant.Conclusion:1.In the pathogenesis of COPD rat model of lung and spleen deficiency,due to the damage of gastrointestinal mucosa,Ghrelin-Obestatin signaling pathway and brain-intestine interaction abnormalities,affect the body’s energy metabolism and nutritional status and participate in the pathogenesis of COPD,which may be one of the pathogenesis of COPD malnutrition.2.The method of reinforcing earth to generate matal could obviously improve the general condition and lung function of COPD rats with deficiency of lung and spleen,and regulate the expression of Ghrelin,Obestatin and their receptors in hypothalamus and duodenum,affects the Ghrelin-Obestatin signaling pathway.3.The method of reinforcing earth to generate matal provide theoretic and experimental evidence for the prevention and treatment of COPD by controlling the Ghrelin-Obestatin signaling pathway and affecting brain-intestine interaction to prevent and treat COPD with malnutrition.