| The first part:TGF-?1/smads Signaling Pathway is Regulated by HSG Gene in COPD Rats' AirwayObjectiveTO explore the correlation of Hyperplasia Suppressor Gene(HSG)with TGF-?1/Smads signaling pathway and to discuss the role of TGF-?1/Smads signaling pathway in airway remodeling of Chronic Obstructive Pulmonary Disease(COPD)and its mechanism,detect whether HSG was involved in COPD airway remodeling by regulating the TGF-?1/smads signaling pathway.MethodThe 34 SPF healthy male SD rats(aged 6 weeks,weighing about 100g)were randomly divided into two group:COPD model group(n=20)and the control group(n=14).Referenceing modeling methods of Song Yiping,the model of COPD rats was induced by lipopolysaccharide plus smoked method.,and the normal groups do not do special treatment.After the success ofbuilding models,each group was randomly selected one rat lung tissue to identify model after putting rats to death by Hematoxylin-Eosin stain-ing(HE stain).The expression of Matrix Metallo Protein-9(MMP-9),Platelet Derived Growth Factor(PDGF),Transforming Growth Factor Beta(TGF-?1)proteins were detected by Enzyme Linked Immunosorbent Assay(ELISA).The expression of HSG?PDGF?TGF-?1?MMP-9?Smad4 mRNA in bronchial tissue of COPD model rats were detected by Real-time Quantitative PCR(RT-PCR).The relationship between Smads with MMP-9,TGF-?1,PDGF was analyzed.The relationship between HSG and PDGF?TGF-??MMP-9?Smad4 was analyzed too.P<0.05 has statistical significance,P<0.01 has significant statistical significance.Results(1)The expression of HSG gene in bronchial tissue of COPD model rats was(0.39±0.06)which was lower than normal group(1.01±0.01),(P<0.01)the expression of HSG protein in bronchial tissue of COPD model rats was(0.50±0.09)which was lower than normal group(1.46±0.07),(P<0.01),with significant statistical significance.The expression of Smad4?MMP-9?TGF-?1 and PDGF genes in bronchial tissue of COPD model rats were(2.02±0.06?1.96±0.04?1.97±0.02?2.04±0.06)severally,which was higher than those in normal group(1.00±0.02?1.02±0.02?1.01±0.02?1.01±0.01),(P<0.01),with significant statistical significance.The expression levels of Smad4?MMP-9?TGF-?1 and PDGF protein in bronchial tissue of COPD model rats were(2.02±0.06?1.96±0.04?1.97±0.02?2.04±0.06)severally,which was higher than those in normal group(1.00±0.02?1.02±0.02?1.01±0.02?1.01±0.01),(P<0.01),with significant statistical significance.(2)HSG gene was negatively correlated with Smad4?MMP-9?TGF-?1?PDGF gene respectively in bronchial tissue of COPD model rats,which r values were-0.889?-0.718?-0.700?-0.784,(P<0.01),with statistical significance.HSG protein was negatively correlated with Smad4?MMP-9?TGF-?1?PDGF protein respectively in bronchial tissue of COPD model rats,which r values were-0.678?-0.669?-0.828?-0.660,(P<0.01),with statistical significance.(3)Smad4 gene was positively correlated with MMP-9?TGF-?1?PDGFgene respectively in bronchial tissue of COPD model rats,which r values were 0.896?0.871?0.929,(P<0.01),with statistical significance.Smad4 protein was positively correlated with MMP-9?TGF-?1?PDGF protein respectively in the bronchial tissue of COPD model rats,which r values were0.757?0.747?0.838,(P<0.01),with statistical significance.Conclusion(1)The expressions of HSG at gene and protein levels in bronchial tissues of COPD rats were lower than those in normal group.The expressions of Smad4?MMP-9?TGF-?1?PDGF at gene and protein levels in bronchial tissues of COPD rats were higher than those in normal group.(2)There was a positive correlation between Smad4 with MMP-9,TGF-?1,PDGF at gene and protein levels in bronchial tissue of COPD rats,which prove that to speed up the release of MMP-9?PDGF?TGF-?1 factor by active TGF-?1/Smads signaling pathway may promotes airway remodeling in COPD rats.(3)It was inferred that HSG could affect the release of MMP-9,PDGF and TGF-?1 by control TGF-?1/Smads signaling pathway because the negative correlation between HSG with MMP-9,TGF-?1,PDGF in bronchial tissue of COPD rats,and it could intervene COPD rat airway remodeling by these ways.The second part:Study on the Correlation Between Smad4 With the Syndromes of TCM in Acute Exacerbation in COPD PatientsObjectiveBy detecting the expression of Smad4 gene which is the iconic gene of TGF-?1/smads signal pathway in the blood of acute exacerbation of chronic obstructive pulmonary disease(AECOPD)patients,to compare the difference of the expression of Smad4 in AECOPD patients with different syndromes of TCM,so as to provide an objective standard for the diagnosis of TCM syndromes.MethodCollect 65 AECOPD patients in the respiratory department of the second affiliated hospital of Guiyang College of Traditional Chinese Medicine(third rate hospital)between March 2017 to November 2017,which were divided into the stagnation of phlegm in lung group(n=20),the invasion of lung by phlegm and heat group,(n=20),the stagnation of phlegm in the brain(n=2),edema syndrome due to Yang deficiency(n=3),the Qi deficiency of lung and kidney group(n=20).And select 15 health people as the control group to observation.Because of the lack of the number of the stagnation of phlegm in the brain and edema syndrome due to Yang deficiency,the sample size was so small,without statistical significance,that was eliminated.To draw the venous blood for the selected patients on an empty stomach,To extract blood RNA with kit and to detected the expression of Smad4 in the blood of different syndromes of TCM in AECOPD patients by Real-time Quantitative PCR(RT-PCR),and to explore the relevance between them.ResultThe expressions of Smad4 gene in the blood of AECOPD patients in stagnation of phlegm in lung group,invasion of lung by phlegm and heat group,Qi deficiency of lung and kidney group respectively were(1.49±0.04),(2.54±0.07),(3.13±0.22),which were higher than those in the normal group(1.26±0.04),(P<0.01),with statistical significance.The expression of Smad4 gene above the each groups:the normal group<the stagnation of phlegm in lung group<the invasion of lung by phlegm and heat group<the Qi deficiency of lung and kidney group(P<0.01),with statistical significance.ConclusionThere was a significant difference in three TCM syndromes groups about the expression of Smad4 gene in TGF-(31/smads signaling pathway.This study can provide an objective standard for the diagnosis of AECOPD. |
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