Clinical Study On The Treatment Of Idiopathic Pulmonary Fibrosis With Pulmonary Vital Granules And Its Regulation Of JAK/STAT Signaling Pathway Expression In Rats With Pulmonary Fibrosis

Objective:To observe the clinical efficacy of Feiweikang granule in the treatment of idiopathic pulmonary fibrosis（Idiopathic pulmonary fibrosis,IPF）,and to further establish an in vivo experiment to explore the regulatory effect of Fei Weikang granule on JAK/STAT signal pathway in rats with pulmonary fibrosis,hoping to provide reference for exploring the mechanism of Feiweikang granule and expanding its clinical application.Methods:（1）Clinical study:64 subjects were enrolled in this study.A randomized controlled study of Fei Weikang granule and pirfenidone was established by random table method.The experimental group was treated with Fei Weikang granule 9g and the control group was treated with pyrifenidone orally.The initial dosage was 200mg.The drug was maintained at the level of 600mg,200mg each time within 2 weeks,and the treatment period was 12 weeks.The effects of Feiweikang granule on TCM clinical syndrome score,6-minute walk test（6-Minute Walking Test,6MWT）and lowest oxygen saturation during a 6MWTL（L-SPO₂）during 6MWT in patients with IPF were observed,and the drug safety and adverse reactions were evaluated.（2）Animal study:after one week of adaptive feeding,48 SD male rats were randomly divided into 6 groups:blank group（NS）,model group（BLM）,pirfenidone group（BF）,low and high dose of Fei Weikang granule group（FH）,Fei Weikang granule medium dose group（FM）and Fei Weikang granule low dose group（FL）,with 8 rats in each group.Except for the NS blank control group,the pulmonary fibrosis model rats were established by intratracheal administration of bleomycin solution at the concentration of 5 mg kg^-1,and the NS group was treated with the same amount of normal saline for blank control.The next day after the completion of the model,normal saline,pirfenidone solution and corresponding concentration of Fei Weikang suspension were given respectively according to the group.After 28 days of intervention,the experimental animals were killed and the samples were collected and analyzed:the detection items were histopathological examination（H&E,Masson staining）,determination of hydroxyproline（Hydrox yproline,HYP）,detection of TGF-β1,IL-6,IL-1βby enzyme linked immunosorbent assay（Elisa）,Western blotting（Western blot,WB）and real-time fluorescence quantitative polymerase chain reaction（Real-time quantitative polymerase chain reaction,RT-PCR）to determine the levels of JAK2,STAT3,α-SMA,Col1 and Fn and the expression of m RNA.Results:（1）Clinical study:after 12 weeks of treatment with Fei Weikang granule,the results of 6MWT test in the test group were better than those in the control group,and the score of clinical syndrome of traditional Chinese medicine decreased significantly compared with the control group.The total effective rate was 65.5%,which was better than that of the control group（27.6%）.And the adverse reaction rate of the test group was only 3.45%,and the difference was statistically significant.（2）In vivo experiment:compared with the blank group,there were more chronic injury and structural damage of lung tissue in BLM group,which was characterized by significant infiltration of inflammatory cells in alveolar space and excessive collagen deposition.The chronic injury,structural damage and collagen deposition of lung tissue in pifenidone group and Feiwei Kang group were significantly less than those in BLM group.It is suggested that both Fei Weikang granule and pirfenidone can reduce the inflammatory infiltration and fibrosis injury such as collagen deposition in lung tissue induced by BLM.The expression levels of TGF-β1,α-SMA and Col1 in lung tissue of BLM group were significantly higher than those of blank group（P<0 01）.At the same time,ELISA detection of inflammatory factors IL-6 and IL-1βalso found the over-expression of inflammatory factors in BLM group.However,the levels of TGF-β1,α-SMA and Col1 in Pirfenidone group and Fei Wei Kang group were significantly lower than those in BLM group,and the corresponding inflammatory factors IL-6 and IL-1βalso showed low expression.In addition,the expression levels of JAK2,STAT3 and IL-6 in lung tissue of rats in BLM group were significantly higher than those in blank group,while the expression levels of JAK2,STAT3 and IL-6 in pifenidone group and Feiwei Kang group were significantly lower than those in BLM group（P<0.05）.Conclusion:Feiweikang granule can improve the TCM clinical syndrome score of IPF patients,alleviate the symptoms of patients,and increase the walking distance of 6MWT and activity tolerance of IPF patients to a certain extent.Fei Weikang granule can improve the activity,reaction sensitivity and mental state of BLM-induced pulmonary fibrosis model rats to a certain extent,diet and other general conditions,and reduce fibrosis injury such as collagen deposition.Decrease the levels of TGF-β1,α-SMA,Col1,Fn and inflammatory factors IL-6 and IL-1β,and then reduce the activation level of myofibroblasts and inflammatory infiltration in lung tissue induced by BLM.In addition,Fei Weikang granule can also inhibit the activation of JAK2/STAT3mediated by IL-6 to some extent,reduce the expression of inflammatory cytokines such as TGF-β1 and IL-1βand the transformation of fibroblasts into myofibroblasts,and reduce the degree of fibrosis and inflammatory infiltration of lung tissue in rats with pulmonary fibrosis induced by BLM.