Exploration Of The Anti-fibrosis Effect Of Dredge Lung Collaterals By Moxibustion Based On TGF-? Signal Pathway

Objective:Pulmonary Fibrosis is a kind of unknown cause,progressive,and fatal pulmonary interstitial disease. TGF-? Played an important role in the process of pulmonary fibrosis. It can not only activate the Smad signaling pathways,induced the downstream epithelial-mesenchymal transition,Also can control various cytokines and aggravate extracellular matrix Real-time.So this topic proposed on the basis of previous research results,the clearlung collaterals treatment,selection (lung shu,kao huang shu) moxibustion,observed by bleomycin copy classical pulmonary fibrosis in rats model of Transforming growth factor-? in moxib ustion therapy treatment.observed the dynamic change of preliminary clarify clear lung collaterals method Transforming growth factor-? key sites and regulation of pulmonary fibrosis.It To improve the prevention and treatment in clinical efficacy about theoretical and experiment.Method:8 weeks health SPF rats 88,weight 180±20g,male,Rats were randomly grouped, normal group (N) and 8;Model group(M):7 days(M7)group and 8,11 days group (M11) and 8,23 days group (M23)and 8,35days group(M35) and 8; Moxibustion group (A):moxibustion 11 days group (A 11)8,moxibustion 23 days group(A23)8,moxibustion 35 days group (A35) and 8;Positive control group (prednisone,P):11 days group (P11) and 8,23 days group (P23)and 8,35days group(P35) and 8;Building endotracheal one-time injection by adopting the method of Thrall to duplicate the pulmonary fibrosis model induced by bleomycin,control group by the method of building same endotracheal one-time injection volume of physiological saline,etc.Each building began treatment and intervention in the next day,alternating moxibustion group bilateral lung shu,kao huang shu paste; Prednisone prednisone group to gavage by Prednisone suspension liquid,All the control group and model group and moxibustion group to adopt the same grab is fixed,not given any treatment.observd thegeneral conditionall rat,And choose 7,11,23,35 days (one day) after each procedure for execute,observed the change EMT and ECM induced TGF-beta signaling pathways in the process of pulmonary fibrosis Dynamical.Each group generally observed in rats,and records breathing,the colour,spirit,the sensitivity and weight,etc.count the lung inx by Right lung tissue and measure HYP lever by ELISA kit.HE and MASSION dye to Observe of moxibustion effect on pulmonary fibrosis model rats in the form of organization.Moxibustion induce the EMT regulation mechanism in lung tissue of pulmonary fibrosis model of rats,Through test the expression of TGF-? in rat lung tissue,Take the organization of the upper lobe for real-time fluorescence quantitative polymerase chain reaction (QPCR) to detect E-cadherin mRNA,alpha-SMA mRNA,vimentin mRNA expression.Explorethe moxibustion's influence on the EMT by TGF-? pathway. Moxibustion regulate the main ingredients of ECM in model of rat lung, test the lever of COL-?,COL-?, HA,LN by ELISA kit,Test the expression of MMP-2/MMP-9,TIMP-?/TIMP-2 by QPCR,Obseve the Protein expression of COL-?,COL-? by IHC,Obseve the key cytokines and key enzymes of metabolism of ECM in rat pulmonary fibrosis model by Moxibustion tretment.Result:1.Moxibustion group in general condition improved substantially. normal group rats is steady breathing,hair moist bright and responsive, normal weight growth; Model group rats shortness of breath,chest and increase faster,the nose green and purple was at onyx spiritual malaise,curled up,outlining the hair sparse lack lustre weight decreased obviously;After treatment,moxibustion group compared with model group generally has a greater improvement;No difference between the positive control group with moxibustion group.2.Lung inx and HYP lever have decreased obviously in Moxibustion group. Model group,the moxibustion group and prednisone group each lung inx were significantly greater than the normal group(P<0.05);model group 23d lung inx and 35d were higher than the moxibustion group23d and 35d,there are no statistical difference (P<0.05).Model group,moxibustion group and prednisone group,the lung tissue hydroxyproline values were significantly greater than the normal group,with statistical significance(P<0.05).Compared with model group, moxibustion group,prednisone group the hydroxyproline values were significantly lower in lung tissue(P<0.05).35 days after,the hydroxyproline value has closest between moxibustion treatment group and normal group in lung tissue.3.Pathological observation found that inflammatory cells infiltration and hyperplasia of collagen fibers decreased significantly in moxibustion group. Normal group of lung tissue structure integrity,thickening of alveolar interval has no edema,inflammation,and fibrosis,no obvious exudation in the alveolar space,compared to normal lung tissue,model group 7d has pulmonary interstitial macrophages and neutrophils cell infiltration,pulmonary interstitial broadening, model group 11 d was alveolar interstitial,Thickening of alveolar walls obviously,a large number of inflammatory cells infiltration.still visible.model group 23d,a large number of inflammatory cells infiltration can be seen in the alveolar interstitial and collagen fiber hyperplasia.model group 35d,a large number of fibers can be seen in the pulmonary interstitial cells.Model group,the moxibustion combined prednisone group degree of alveolitis and pulmonary fibrosis did badly in the normal,the lever of collagen deposition less than moxibustion group 35d and prednisone group.4.Moxibustion Inhibition the ECM in lung tissue of pulmonary fibrosis in rats model. The TGF-? content in 11d up to a peak in Pulmonary fibrosis rats lung tissue,compared with normal group and had significant difference(P<0.05).In Moxibustion group and prednisone group,TGF-P content has significantly reduce,moxibustion group at 11d and 23d compared to model group there were significant differences (P<0.05).Compared with the normal group, model group 7d, the mRNA expression of E-cad quantity increased obviously,there were significant differences (P<0.05);especially compared to the model group,moxibustion group 11d and 23d E-cad mRNA expression,there were significant differences(P<0.05);the E-cad mRNA expression is between Model group and moxibustion group in the opposite trend,Model group and moxibustion group of alpha-SAM and vimentin mRNA expression trend is roughly similar,From 11 to 23 days on the rise,and 23 days to 35 days is on the decline.5.Moxibustion reduce the main ingredients of ECM in model of rat. lungModel group 7d the level of HA and LN in the lung tissue obviously higher than normal group,and there are significant statistical significance(P<0.05),after 35 days,the levels of HA and LN in moxibustion treatment group are lower than the model group.Model group rats lung tissue COL-?,COL-? content with the passage of time is roughly the trend of rise. compared with the normal group,model group 7d COL-?,COL-? levels were significantly elevated,has a very significant statistical significance(P<0.05),Compared with model group, moxibustion group,prednisone treatment group at treatment stage of rat lung COL-?,COL-? content decreased in 11 and 35 days,with statistical significance(P<0.05).The MMP-2 and MMP-9 mRNA of expression in Model group,the moxibustion group at the same time are roughly the same trend.MMP-2 mRNA expression of from 11 days to 23 days showed a trend of rising,and declined to 23 days in Model group and moxibustion group.Model group MMP-9 mRNA expression from 11 days to 35 days showed a trend of rising.moxibustion group in 11 days and 23 days,compared with model group MMP-9 mRNA expression, had statistical significance(P<0.05).TIMP-1 mRNA expression of the moxibustion group and Model group are roughly same trend at the same point,the TIMP-1 mRNA expression of Model group and moxibustion group,showed a trend of rising from 11 days to 23 days,and 23 days to 35 days is decline.11 days moxibustion and prednisone treatment group compared with model group,had statistical significance(P<0.05).the TIMP-2 mRNA expression of moxibustion groupand and Model group is different at change trend the same time.the TIMP-2 mRNA expression of Model group from 11 to 23 days continue to rise,and 23 to 35 days is decline.TIMP-2 mRNA expression of Moxibustion group and prednisone group from 11 days to 35 days showed a trend of rising.11 days and 23 days,TIMP-2 mRNA expression of moxibustion and prednisone treatment group compared with model group was statistically significant(P<0.05).Conclusion:Dredge lung collaterals moxibustion can improve the overall situation in the rat model of pulmonary fibrosis model,Reduce alveolar inflammation and the lung tissue damage,Inhibition of TGF-beta raised and delay the EMT process,To prevent lung extracellular matrix formation,Adjust the imbalance MMPs/TIMPs,Multi-angle level to delay the process of pulmonary fibrosis,Fully embodies the dredge lung collaterals moxibustion integrity network medicine characteristics and advantages of prevention and treatment of pulmonary fibrosis.