| Objective:1 By studying the expression characteristics of PD-1、PD-L1 and other related immune indicators in COPD phlegm-turbid obstructing lung syndrome,to explore the characteristics of COPD phlegm-turbid obstructing lung syndrome,in order to find the internal relationship between the characteristics of COPD phlegm-turbid obstructing lung syndrome and PD-1、PD-L1 and other immune indicators,to provide objective basis for TCM clinical syndrome differentiation.2 By observing the expression characteristics of general laboratory indicators,CRP,IL-6,McP-1,MIP-1α,TNF-α and other inflammatory factors in COPD phlegm-turbidity obstructing lung syndrome,to explore the characteristics of COPD phlegm-turbidity obstructing lung syndrome,in order to find the internal relationship between the characteristics of COPD phlegm-turbidity obstructing lung syndrome and modern laboratory indicators,inflammatory indicators.To provide objective basis for TCM clinical syndrome differentiation.3 By establishing a COPD mouse model,the expression characteristics and possible regulatory mechanisms of PD-1、PD-L1 and other related immune molecules in COPD were explored based on the PD-1/PD-L1 axis,and the influence on the expression of related indicators and possible mechanism of action were discussed after the intervention of Maxingloushi decoction,which has guiding significance for further clinical and research.To provide objective basis for clinical treatment of COPD.Methods:1 The clinical study was conducted in a cross-sectional epidemiological study 65 patients with COPD were collected according to the inclusion and exclusion criteria,and analyzed related immune indexes such as PD-1 and PD-L1,inflammatory indexes such as CRP,IL-6,MCP-1,MIP-1α and TNF-α,as well as general laboratory indexes such as CAT score,mMRC score,FeNO test and lung function test.To explore the expression characteristics of observation indexes in COPD phlegm-turbidities obstructing lung syndrome.2 In the experimental study,the animal model of COPD was constructed and divided into four groups,namely,Maxingloushi decoction group(TCM group),normal group,PD-1 inhibitor group(control group)and COPD model group(model group).Different intervention measures were given respectively.The expression characteristics of PD-1,PD-L1,IL-6,MCP-1,MIP-1α and TNF-α were detected by ELISA.The expression of NF-κB in lung tissues of COPD model mice was analyzed by Western Blot.Flow cytometry was used to analyze the expression characteristics of PD-1 and PD-L1 in spleen,whole blood and alveolar lavage fluid of COPD model mice,to explore the expression characteristics of PD-1,PD-L1 and other related immune molecules in COPD and the possible regulatory mechanism,and to explore the influence of Chinese traditional medicine Maxingloushi decoction on the expression of detection indicators and the possible mechanism of action.Results:1 Expression characteristics of laboratory indicators in COPD phlegm-turbidities obstructing lung syndrome:The expression of PaCO2 in blood gas analysis of COPD patients with phlegm-turbidities obstructing lung was lower than that of patients with lung-kidney qi deficiency syndrome(P<0.05).The expressions of WBC,PCT,PLR and TT4 in COPD patients with phlegm-turbidity obstructing lung were higher than those in patients with lung-kidney qi deficiency(P<0.05).The expression of FEV1 and FEV1/FVC in COPD patients with phlegm-turbidity obstructing lung was higher than that in COPD patients with lung-kidney qi deficiency(P<0.05).The expression of FeNO index in COPD patients with phlegm-turbidness obstructing lung was slightly higher than that in patients with lung-kidney qi deficiency(P>0.05),there was no significant difference in CAT score,mMRC score and other laboratory indicators between the two syndromes(P>0.05).2 The expression characteristics of inflammatory indicators in COPD patients with phlegm-turbidities obstructing lung syndrome were as follows:the expression of MCP-1 in COPD patients with phlegm-turbidities obstructing lung was higher than that in patients with lung-kidney qi deficiency syndrome(P<0.05),MIP-1α was slightly lower in COPD patients with phlegm-turbidities obstructing lung than in lung-kidney qi deficiency syndrome(P>0.05).The expression of MCP-1 and MIP-1α in COPD patients with phlegm-turbidity obstruction of lung and deficiency of lung-kidney Qi was higher than that in normal control group(P<0.05).The expression of ESR,IL-6 and CRP in COPD patients with phlegm-turbidity obstructing lung was higher than that in COPD patients with pneumon-kidney qi deficiency(P<0.05).3 The expression characteristics of immune indexes in COPD patients with phlegm-turbidity obstructing lung syndrome:the expression of PD-1 in COPD patients with phlegm-turbidity obstructing lung syndrome was lower than that in patients with lung-kidney qi deficiency syndrome(P<0.05),the expression of PD-L1 was higher than that of patients with lung-kidney qi deficiency syndrome(P<0.05).The expression of PD-1 in COPD patients with phlegm-turbidities and lung-kidney qi deficiency was higher than that in healthy volunteers(P<0.05),PD-L1 expression in COPD patients with phlegm-turbidities obstructing lung and pneumon-kidney qi deficiency was lower than that in healthy volunteers(P<0.05).The IgE expression in COPD patients with phlegm-turbidity obstructing lung was higher than that in patients with lung-kidney qi deficiency(P<0.05),CD3+expression in COPD patients with phlegm-turbidity obstructing lung was higher than that in patients with lung-kidney qi deficiency(P<0.05);IgM,IgA,IgG,C3,C4,CD3+CD4+,CD3+CD8+,CD4+/CD8+showed no significant difference between the two syndroms(P>0.05).4 Experimental research:① Lung tissue HE staining:There were abnormal structures in the lung tissues of COPD model mice,including alveolar atrophy and collapse,alveolar wall thickening,and neutrophil infiltration.The pathological manifestations of lung tissue in Maxingloushi decoction group and the PD-1 inhibitor group were improved after drug intervention.②ELISA detected MCP-1,MIP-1α,IL-6 and TNF-α:The levels of MCP-1 and MIP-1αin plasma and alveolar lavage fluid of model group were higher than those of normal group,Maxingloushi decoction group and control group(P<0.05),and the levels of plasma MCP-1 and MIP-1α in Maxingloushi decoction group were lower than those in control group(P<0.05),the expression of MIP-1α in Chinese herbal group was lower than that in control group(P<0.05),the expression of MCP-1 in alveolar lavage fluid in Maxingloushi decoction group was slightly lower than that in control group(P>0.05).The levels of IL-6 and TNF-α in plasma and alveolar lavage fluid of model group were higher than those of normal group,Maxingloushi decoction group and control group(P<0.05),and the levels of IL-6 and TNF-αin plasma and alveolar lavage fluid in Maxingloushi decoction group were lower than those in control group(P<0.05).③Lung tissue NF-κB was detected by Western Blot:the expression of NF-κB in model group was higher than that in Maxingloushi decoction group,control group and normal group(P<0.05),and the expression of NF-κB in lung tissues of Maxingloushi decoction group was lower than that of control group(P<0.05).④IHC staining and IOD value in lung tissue:The expression of PD-1 in model group was higher than that in normal group(P<0.05),PD-L1 expression in model group was lower than that in normal group(P<0.05).The expression of PD-1 in control group was lower than that in model group,and the expression of PD-L1 was higher than that in model group(P<0.05).The expression of PD-1 in Maxingloushi decoction group was lower than that in model group and control group(P<0.05),the expression of PD-L1 in Maxingloushi decoction group was higher than that in model group and control group(P<0.05).⑤PD-1 and PD-L1 in plasma and alveolar lavage fluid were detected by ELISA:The expression of PD-1 in plasma in model group was higher than that in normal group,Maxingloushi decoction group and control group(P<0.05),the plasma PD-L1 expression in model group was lower than that in normal group,Maxingloushi decoction group and control group(P<0.05),and plasma PD-1 expression in Maxingloushi decoction group was lower than that in control group,and plasma PD-L1 expression was higher than that in control group(P<0.05).The expression of PD-1 in alveolar lavage fluid in model group was higher than that in normal group,Maxingloushi decoction group and control group(P<0.05),the expression of PD-L1 in alveolar lavage fluid in model group was lower than that in normal group,Maxingloushi decoction group and control group(P<0.05),and the expression of PD-1 in alveolar lavage fluid in Maxingloushi decoction group was lower than that in control group,and the expression of PD-L1 in alveolar lavage fluid in Maxingloushi decoction group was higher than that in control group(P<0.05).⑥Flow cytometry detected PD-1 and PD-L1 in spleen,blood and alveolar lavage fluid:The expression of PD-1+CD4 cells in spleen and whole blood in the model group was higher than that in normal group,Maxingloushi decoction group and control group(P<0.05),the proportion of PD-1+CD4 cells in T cells in alveolar lavage fluid in model group was higher than that in normal group and Maxingloushi decoction group(P<0.05).The expression of PD-1+CD8 cells to T cells in alveolar lavage fluid in model group was higher than that in normal group,Maxingloushi decoction group and control group(P<0.05),there was no significant difference in the ratio of PD-1+CD8 cells to T cells in spleen and whole blood of model group(P>0.05).The expression of PD-1+CD4 cells in spleen,whole blood and alveolar lavage fluid in model group was higher than that in normal group,Maxingloushi decoction group and control group(P<0.05).The expression of PD-1+CD8 cells in alveolar lavage fluid in model group was higher than that in normal group,Maxingloushi decoction group and control group(P<0.05),the expression of PD-1+CD8 cells in spleen in the proportion of total lymphocytes in model group was higher than that in normal group(P<0.05)and there was no significant difference in other groups(P>0.05),there was no significant difference in the expression of PD-1+CD8 cells in the proportion of total lymphocytes in model group(P<0.05).The expression of MHCII+M1 Macrophage PD-L1+in spleen,whole blood and alveolar lavage fluid of model group was higher than that of normal group and Maxingloushi decoction group(P<0.05).Conclusion:1 General laboratory indicators,such as WBC,PCT,PLR,PaCO2,TT4,lung function;inflammatory indicators,such as MCP-1,MIP-1α,IL-6,ESR,CRP;and immune indicators,such as PD-1,PD-L1,IgE,CD3+,have corresponding expression characteristics in COPD patients with phlegm-turbide-obstructed lung syndrome.At the same time,compared with the patients with lung-kidney qi deficiency syndrome,it was found that the inflammatory response and immune response of COPD patients with phlegm-turbidities obstructing lung were more obvious,while the immune damage was less severe than that of the patients with lung-kidney qi deficiency syndrome.It indicates that the characteristics of the syndrome of COPD phlegm-turbidium-obstructing lung have internal relationship with modern laboratory indicators,inflammatory indicators and immune-related indicators,suggesting that these observation indicators have certain guiding significance in COPD TCM syndrome differentiation,and provide a basis for TCM clinical syndrome differentiation.It can also be used as an auxiliary reference index for accurate syndrome differentiation and classification between the two syndromes of COPD:phlegm-turbidity obstructing lung and deficiency of lung-kidney qi.2 There were obvious inflammatory reactions in lung tissues of COPD model mice.COPD can significantly increase the expression of inflammatory factors MCP-1,MOP-1α,IL-6 and TNF-α,as well as NF-κB P65 protein.Both Maxingloushi decoction and PD-1 inhibitor can improve the inflammatory response of COPD,significantly reduce the expression of inflammatory factors MCP-1,MIP-1α,IL-6 and TNF-α,and down-regulate the expression of NF-κB P65 protein,and the improvement effect of Maxingloushi decoction is better than that of PD-1 inhibitor.It is suggested that Maxingloushi decoction can significantly improve the inflammatory response of COPD model mice.3 The PD-1/PD-L1 axis plays an important role in the obvious immune response in the lung tissues of COPD model mice.COPD can induce the high expression of PD-1 in lung tissue,plasma and alveolar lavage fluid of mice,and inhibit and down-regulate the expression of PD-L1.However,Maxingloushi decoction and PD-1 inhibitor can significantly reduce the high expression of PD-1 induced by smoking combined with LPS,and improve the expression of PD-L1,and the effect of Maxingloushi decoction is better than that of PD-1 inhibitor.It is suggested that Maxingloushi decoction can improve the immune impairment of COPD model by regulating The PD-1/PD-L1 axis,regulating the expression of PD-1 and PD-L1 immune molecules,down-regulating PD-1 and up-regulating PD-L1,thus improving the immune response,improving the immune function of the body,playing the role of immune regulation,and improving the immune imbalance of COPD.4 Through clinical and experimental studies,the expression characteristics of PD-1 and PD-L1 in COPD were found,revealing the correlation between the PD-1/PD-L1 axis and the immune mechanism of COPD,and blocking the PD-1/PD-L1 axis can be used as a new target for the treatment of COPD.In this study,it was found that both Maxingloushi decoction and PD-1 inhibitor could regulate the immune function of COPD by blocking the PD-1/PD-L1 axis,and Maxingloushi decoction had a better effect,providing a new treatment idea for further application of Traditional Chinese medicine to improve the immune function of COPD. |
PDF Full Text Request