Effects Of Yufeiyin Decoction And Its Disassembled Prescription On MMP-9 And TIMP-1 Expression In Pulmonary Fibrosis Rats

Objective: To observe the effect of Yufeiyin decoction and its decomposed formula on the expression of matrix metalloproteinase 9(MMP-9)and tissue inhibitor of metalloproteinase 1(TIMP-1)in rats with pulmonary fibrosis induced by bleomycin,thus exploring the effect of this medicine on the pathological process of pulmonary fibrosis and its possible mechanism.Methods: A total of 180 healthy male rats of Specific pathogen Free,weighing 180-200 g,were housed in a clean grade animal room.After 1 week of adaptive feeding,rat model was established by intratracheal instillation of bleomycin.Then,the rats were randomly divided into 6 groups: sham operation group,model group,Yufeiyin group,Buqitongluo group,Yangyintongluo group and Quyutongluo group,with 30 rats in each group.On the 2nd day after surgerying,corresponding dosage menicine was given to the rats(Yufeiyin Group: 12.4 g/kg/d,Buqitongluo group: 5.2 g/kg/d,Yangyintongluo group: 5.1 g/kg/d,Quyutongluo group: 5.3 g/kg/d).The sham operation group and the model group were given normal saline(10 mL/kg),once a day.Body weight,food intake and general state were observed.On the7 th,14th and 28 th day of the experiment,10 rats in each group were selected and anesthetized with sodium pentobarbital(4 mg/100 g)and then decapitated.The lung was taken out and HE combined with Masson staining were applied to observe the pathological change.Immunohistochemistry,Real time fluorescence quantitative PCR(RT-qPCR)and Western-blot were conducted to detect gene and protein expression of MMP-9 and TIMP-1.Results:1.General conditions of ratsIn the sham operation group,the food intake and body weight increased steadily.The skin and hair was smooth,and the breathing was stable.In themodel group,the rats were weak,lazy in activities,with shortness of breath.The food intake and body weight decreased first,followed by a solwly growth.2.Pathological changes in lung tissueIn the sham operation group,the alveolar structure was intact.No structural disorder,inflammatory cell infiltration and collagen deposition were observed.In the model group,the alveolar septum was thickened and the alveolar was in different size,meanwhile,there were a large number of inflammatory cell exudation and collagen fiber formation.3.Expression of MMP-9 and TIMP-1 genes and proteinsCompared with the sham operation group,the expression of MMP-9 and TIMP-1 mRNA and protein in the lung tissues of the model group have significantly difference(P<0.01).Compared with the model group,the expression of MMP-9 mRNA and TIMP-1 mRNA and protein in the lung tissue of rats in each treatment group have statistically significance(P<0.05 or P<0.01),and the degree of improvement of the MMP-9 and TIMP-1 in 3decomposition group was not as good as that of the Yufeiyin group(P<0.05 or P<0.01).Compared beteween 3 decomposition group,the regulation of MMP-9 and TIMP-1 expression in the lung tissues and their ratio of Buqitongluo group were superior to Yangyintongluo group and Quyutongluo group(P<0.05 or P<0.01).Conclusion:1.Qi deficiency,Yin deficiency and blood stasis are important syndromes of mediated the pathological process of pulmonary fibrosis,and Qi deficiency is the most critical pathogenic factor throughout the disease.Benefiting Qi and nourishing Yin,removing blood stasis and dredging collaterals can be used as the conventional treatment for pulmonary fibrosis,especially to benefiting Qi is the key to treat the disease.2.Yufeiyin and its decomposition prescriptions can defer the pathological changes of pulmonary fibrosis,the mechanism may be related to the inhibition of MMP-9,TIMP-1 and balance the ratio.