| The first part:study on the correlation between ERK1/2 and TCM syndromes of AECOPD patientsObjective:Through observing the expression level of ERK1/2(ertracellular-signal regulated kinase 1/2)genes in different TCM syndromes of patients with AECOPD(acute exacerbation of chronic obstructive pulmonary disease),which is the marker gene of the ERK-signal-path(ertracellular-signal regulated kinase),to explore the roles of ERK-signal-path in airway remodeling of AECOPD and try to find AECOPD TCM syndrome differentiation from the genetic level.The methods:According to The TCM Type of AECOPD,We divided 60 patients with AECOPD into four groups and then divided into three groups,group of Phlegm blocking in lung,group of Retention of Heat-Phlegm in Lung and group of Phlegm-Accumulation in Lung,While selected 20 normal human as control group.Application of RT-PCR in discovering the expression level of ERK1/2?HSG genes(hyperplasia suppressor gene)in peripheral blood of all groups,and to explore the clinical significance and difference of ERK1/2,HSG gene in the three TCM syndromes in AECOPD.Statistics:The data were analyzed by SPSS 16.0.The measurement data are expressed as mean ± standard deviation(x±s),The measurement date in line with normal distribution used One-Way ANOVA,While,used the nonparametric test.Results:The expression of HSG gene in serum of patients with AECOPD was as follows:the level of HSG gene in the normal control group was 0.588 ± 0.069,which was significantly higher than that in the Phlegm blocking in lung group(0.542 ± 0.031),Retention of Heat-Phlegm in Lung group(0.537±0.030)and group of Phlegm-Accumulation in Lung(0.518±0.024),The difference was statistically significant(P<0.05).The expression of ERK1/2 gene in serum of patients with AECOPD was as follows:the Phlegm blocking in lung group(2.013±0.377),Retention of Heat-Phlegm in Lung(2.164±0.350)and group of Phlegm-Accumulation in Lung(2.330±0.298).The three groups were higher than that in the normal control group,which was(1.108±0.355).The difference was statistically significant(P<0.05).Conclusions:The relative expression of ERK1/2 mRNA was gradually increased from Phlegm blocking in lung group,Heat-Phlegm in Lung group and Phlegm-Accumulation in Lung group and HSG gene gene is the opposite,Thus,those provided some experience for the quantification and diagnosis of AECOPD TCM syndrome differentiation.The second part:HSG regulates ERK signaling pathway to inhibit airway remodeling in COPD ratsObjective:Lay the foundation for Genetherapy in COPD by investigating the mechanism of airway remodeling of COPD.by interfering the hyperplasia suppressor gene(HSG).The methods:50 Male SD rats were divided randomly into COPD group(n = 40)and control group(n = 10),and we established COPD model by using the method of smoking cigarettes and injecting Lipopolysaccharide into trachea of rats.Meanwhile,we used tissue adherent method to culture airway fibroblasts,and discovered the expression level of HSG?ERK1/2 in airway fibroblasts by Western blot,discovered the mRNA expression of them by RT-PCR,At last,we used ELISA to detect the levels of TGF-?1,PDGF,MMP-9 in supernatants of airway fibroblasts.Statistics:The data were analyzed by SPSS16.0.The measurement data are expressed as mean ±standard deviation(x±s),The measurement date in line with normal distribution used One-Way ANOVA,While,used the nonparametric test.Results:The expression of ERK 1/2 protein in airway fibroblasts of COPD rats was0.591±0.099,the expression of ERK 1/2 protein in airway fibroblasts of normal group was 0.269 ±0.061,and ERK1/2 in airway fibroblasts of COPD rats protein expression was higher than that of normal rats(P<0.05).The expression of HSG protein in airway fibroblasts of normal rats was 0.649 ± 0.080,and the expression of HSG protein in airway fibroblasts of COPD group was 0.287± 0.112.The expression of HSG protein in airway fibroblasts of COPD rats was higher than that in normal rats Low(P<0.01).The expression level of PDGF protein in supernatant of COPD rats was 26.49 ±0.44,the expression of TGF-?1 protein was 82.50 ± 1.23,the expression of MMP-9 protein was 12.04 ± 0.75,the supernatant of superflora of COPD rats The expression of protein in the three groups was higher than that in the normal group,the difference was statistically significant(P<0.01).The expression level of PDGF gene in the airway fibroblasts of COPD rats was 2.046±0.151,the expression of TGF-?1 was 1.970±0.018,the expression of MMP-9 was 1.758±0.389,the expression of ERK1/2 was 2.036±0.174,The expression of all three genes in airway fibroblasts of COPD rats was higher than that in normal group,the difference was statistically significant(P<0.05).The relative expression of HSG gene mRNA in airway fibroblasts of COPD rats was negatively correlated with the relative expression of MMP-9 mRNA,and the correlation coefficient r=-0.485,P<0.05;HSG gene mRNA in airway fibroblasts of COPD rats was negatively correlated with the relative expression of PDGF mRNA,and the correlation coefficient r=-0.519,P<0.05;HSG in airway fibroblasts of COPD rats The relative expression of HSG gene mRNA and the relative expression of TGF-?1 mRNA in airway fibroblasts of COPD rats were significantly correlated with the relative expression of TGF-?1 mRNA And the correlation coefficient r=-0.519,P<0.05.The relative expression of HSG gene mRNA in airway fibroblasts of COPD rats was negatively correlated with the relative expression of ERK1/2 mRNA,and the correlation coefficient r=-0.617,P<0.05.Conclusions:The expression of ERK 1/2 in airway fibroblasts of COPD rats was higher than that of normal rats,and the expression of HSG was decreased.There was a linear negative correlation between HSG gene and the expression of MMP-9,PDGF,TGF-?1 and ERK1/2 mRNA expression in airway fibroblasts of COPD rats.Thus,we can conclude that HSG could regulate the phosphorylation of ERK1/2 and inhibit the expression of MMP9,TGF-?1 and PDGF expression,than involved in airway remodeling in COPD rats. |
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