HSG Gene Regulates Airway Wnt/PCP Signaling Pathway In COPD Rats And The Correlation Between RhoA Gene And TCM Syndromes In Acute Exacerbation Of COPD

Animal experimentsObjective:Explore HSG(Hyperplasia Suppressor Gene)whether through planar cell polarity pathway,that Wnt/PCP signaling pathway inhibition airway remodeling in COPD(Chronic Obstructive Pulmonary Disease)rats?If HSG gene can,we will ex-plore the mechanism?Methods:The 100 healthy male SD rats were randomly divided into two group:COPD model group and the control group,control group 30,COPD model group 70 rats.Reference modeling methods of Song Yiping,After 28 days,after the two groups of rats were sacrificed,take small airway to culture fibroblasts primary cell,the third generation of cell lines for subsequent experiments.After modeling,each group was randomly selected one rat lung tissue to identify model by Hematoxylin-Eosin staining(HE stain).Using Enzyme-Llinked ImmunoSorbent Assay(ELISA)to detect the expression of Matrix Metalloprotei-9(MMP-9),Platelet Derived Growth Factor(PDGF)and Transforming Growth Factor Betal(TGF-β1)in the small airways into the fibroblast;By Western Blot(WB)to detect the expression of the HSG,RhoA,Daaml protein in fibroblast;By Real-time Quantitative Polymerase Chain Reaction(q-PCR)to detect the mRNA expression of the HSG,PDGF,TGF-β1,RhoA protein in fibroblast,to explore the correlation between HSG gene and MMP-9,TGF-β1,PDGF,RhoA,Daaml.Results:1.Compared with normal group,the concentration of MMP-9,PDGF,TGF-β1 in fibroblast supernatant with COPD model group were significantly increased(▲P<0.01),had statistic significance obviously.2.Compared with normal group,the expression of HSG protein in fibroblast of COPD model group was decreased significantly(**P<0.01),had statistic significance obviously;The expre-ssion of RhoA and Daaml protein in fibroblast of COPD model group was increased significantly increased(*P<0.01),had statistic significance obviously.3.Compared with normal group,the relative expression of HSG gene mRNA in fibroblast of COPD model group was decreased significantly(*P<0.01),had statistic significance obviously;The relative expression of PDGF,TGF-β1 and RhoA gene mRNA in fibroblast of COPD model group was significantly increased(*P<0.01),had statistic significance obviously.4.The expression of HSG,protein was negatively correlated with that of MMP-9,TGF-β1,PDGF,RhoA,Daaml which had statistic significance obviously(P<0.01).Conclusions:There was a linear negative correlation between HSG and MMP-9,TGF-β1,PDGF,RhoA,Daamlprotein expression,therefore,by regulating the expressi-on of HSG may be able to inhibit the Wnt/PCP signaling pathway is abnormally activated,thereby inhibiting the MMP-9,PDGF,TGF-β1 over-expression,which may influence airway remodeling in COPD rats.Clinical experimentsObjective:On the basis of animal studies,attempt to provide an objective standard for the diagnosis of COPD TCM syndrome during acute exacerbation period from the genetic level.Methods:Collect 100 acute exacerbation of COPD patients which were divided into phlegm obstruct the lung,phlegm-heat obstructing lung syndrome,syndrome of orific-es confused by phlegm,deficiency of pulmonary and renal Qi,edema due to yang deficiency,a total of five groups,20 people each group.15 people in control group.All patients were fasting hemospasia,use kit to extracte blood total RNA,operate accor-ding to instructions,by Real-time quantitative polymerase chain reaction to detect the mRNA expression of RhoA gene in blood of patients with COPD TCM syndrome type during acute exacerbation period and to explore the correlation.Results:In addition to the group phlegm-heat obstructing lung syndrome compared with group syndrome of orifices confused by phlegm(*△P =0.288>0.05),there was no difference and no statistically significant.The mRNA expression of RhoA gene in phlegm obstruct the lung,phlegm-heat obstructing lung syndrome,syndrome of orific-es confused by phlegm,deficiency of pulmonary and renal Qi,edema due to yang deficiency were significantly higher than the normal group(P<0.01)and comparison between groups which had statistic significance obviously(P<0.01).The mRNA expression of RhoA gene in five groups TCM syndromes:The phlegm obstruct the lung<phlegm-heat obstructing lung syndrome/syndrome of orifices confused by phlegm<deficiency of pulmonary renal Qi<edema due to yang deficiency,.The mRNA expression of RhoA gene was positive correlated with that of COPD five groups TCM syndrome which had statistic significance obviously(P<0.01).Conclusions:The mRNA relative expression of RhoA gene there are differences in the five groups of TCM syndrome type which can provide some objective diagnostic criteria for acute exacerbation of COPD TCM syndrome during acute exacerbation period and reveal their disease severity in the Wnt/PCP signaling pathway.