| ObjectiveTo observe the distribution characteristics of traditional Chinese medicine syndromes of rheumatoid arthritis-related interstitial lung disease(RA-ILD),and analyze the correlation between RA-ILD traditional Chinese medicine syndromes and some serological indicators.To elucidate the mechanism of Bushentongluo Decoction in intervening with RA-ILD model rats induced by complete Freund’s adjuvant.Methods1 Clinical partThis study used a cross-sectional study approach.A total of 92 patients with RA-ILD who met the inclusive criteria were included in the study.The basic data,four-diagnosis information,and serological indicators such as inflammation and immunity were recorded.We analyzed the clinical data of patients,summarized the distribution characteristics of TCM syndrome types,and analyzed the relationship between syndrome types and related serological indicators.2 Experimental partSixty rats were randomly divided into a blank control group,a model group,a Bushen Tongluo Decoction high-,medium-and low-dose group,and a pirfenidone group.In addition to the blank control group,all the other rats were treated with RA-ILD rat model induced by complete Freund’s adjuvant.The rats were given the corresponding drugs by gavage the next day after modeling,and were intervened for a total of 90 days.At the end of the drug intervention,the arthritis index(AI)scores of the rats were evaluated,and serum and lung tissue specimens were collected and stained with HE and Masson to observe the histopathological changes in the lung tissue.The relative expression levels of p-JAK2/STAT2 and p-STAT3/STAT3 proteins in lung tissues were measured by immunohistochemistry.Results1 Clinical studies(1)A total of 92 patients with RA-ILD were included in this study,of which the majority were female,with a predominant age distribution of 61-70 years.(2)Distribution of RA-ILD evidence types:according to the composition ratio from highest to lowest,they are Lung and Kidney Qi Deficiency,Lung-Loop Paralysis,Lung and Kidney Qi Deficiency,Phlegm and Heat Congestion,Lung and Kidney Qi Deficiency,Phlegm and Heat Congestion,Yin Deficiency and Internal Heat,Lung-Loop Paralysis,Phlegm and Cloudy Lung,Lung Qi Deficiency and Lung and Kidney Qi and Yin Deficiency,the most frequent evidence type is Lung and Kidney Qi Deficiency and Lung-Loop Paralysis.(3)Relationship between immunoglobulins and evidence types:IgA:Compared with yin deficiency and internal heat syndrome,the IgA level of lung-kidney qi deficiency syndrome and lung collateral obstruction syndrome decreased significantly(p<0.05),but there was no significant difference among other syndromes.IgM:Compared with yin deficiency and internal heat syndrome,the IgM level of lung and kidney qi deficiency syndrome,lung and kidney qi deficiency syndrome and lung collateral obstruction syndrome decreased significantly(p<0.05),and there was no significant difference among other syndromes.IgG:Compared with the syndrome of yin deficiency and internal heat,the IgG level of the syndrome of phlegm-heat obstructing the lung was significantly increased(p<0.05).The IgG level of the syndrome of lung and kidney qi deficiency,the syndrome of lung and kidney qi deficiency,the syndrome of lung collateral obstruction,and the syndrome of lung and kidney qi deficiency and phlegm-heat obstructing the lung were also significantly increased(p<0.01).There was no significant difference among the other syndrome types.(4)Relationship between complement and evidence type:Complement C3:There was no significant difference between each syndrome type and complement C3(p>0.05).Complement C4:There was no significant difference between each syndrome type and complement C4(p>0.05).(5)Relationship between autoantibodies and evidence types:Anti-CCP antibody:Compared with the syndrome of lung and kidney qi deficiency,the anti-CCP antibody levels in the syndrome of yin deficiency and internal heat,the syndrome of lung and kidney qi deficiency and phlegm heat obstructing lung were significantly increased(p<0.05).The anti-CCP antibody levels in the syndrome of lung collateral obstruction were also significantly increased(p<0.01).Compared with the syndrome of deficiency of both lung and kidney qi and yin,the anti-CCP antibody levels of the syndrome of yin deficiency and internal heat,the syndrome of lung collateral obstruction,the syndrome of lung and kidney qi deficiency and the syndrome of lung collateral obstruction were significantly increased(p<0.05),and there was no significant difference among the other syndrome types.RF:There was no significant difference between the four syndromes in RF(p>0.05).ASO:There was no significant difference among the four syndromes of ASO(p>0.05).(6)Relationship between the inflammatory indexes and the evidence types:ESR:There was no significant difference in ESR among the syndromes(p>0.05),but the ESR level of the syndromes of lung-kidney qi deficiency and lung collateral obstruction tended to increase compared with other syndromes.CRP:There is no significant difference in CRP among the syndrome types(p>0.05),but the CRP level in the syndrome of lung and kidney deficiency and lung collateral obstruction tends to increase compared with other syndrome types.2 Experimental part(1)Arthritis index(AI)scores in rats:After 28 days of modeling,compared with the blank control group,the AI scores of all other groups increased(p<0.01),and there was no statistically significant difference between the AI scores of the model group,the high,medium and low dose groups of Bushentongluo formula,and the pirfenidone group(p>0.05).After 90 days of administration,compared with the blank control group,the AI scores of all the remaining groups increased(p<0.01);compared with the model group,the AI scores of rats in the high,medium and low dose groups of Bushentongluo formula and Pirfenidone group were significantly lower(p<0.01);compared with the Pirfenidone group,the AI scores of rats in the high,medium and low dose groups of Bushentongluo formula were significantly lower(p<0.01);compared with the three dose groups of Bushentongluo formula,the AI scores of rats in the high,medium and low dose groups of Bushentongluo formula were significantly lower(p<0.01).In the comparison between the three dose groups of Bushentongluo Formula,the AI score of the high dose group was significantly higher than that of the low dose group of Chinese medicine(p<0.05),and the AI score of the medium dose group of Bushentongluo Formula was significantly higher than that of the low dose group of Bushentongluo Formula(p<0.01).(2)HE and Masson staining:no obvious inflammatory cell infiltration was seen in the alveolar interstitium and alveolar cavity of the blank control group.In the model group,the alveolar structure of the rats was destroyed and a large number of inflammatory cells were seen in the alveolar cavity and interstitial tissue,and a large area of obvious fibrous tissue proliferation was seen in the lung tissue;the Bushentongluo Formula group could inhibit the inflammatory cell infiltration and collagen fiber deposition to different degrees.(3)Serum IFN-γ levels:compared with the blank group,serum IFN-γ levels were decreased in all other groups(p<0.01);compared with the model group,serum IFN-γ levels were increased in the low dose group and pirfenidone group of Bushentongluo Formula(p<0.05);compared with the pirfenidone group,serum IFN-γ levels were decreased in the high and medium dose groups of Bushentongluo Formula(p<0.05);compared with the three dose groups of Bushentongluo Formula,serum IFN-γ levels were lower than those in the low dose group of Bushentongluo Formula(p<0.05).Compared with the three dose groups,the serum IFN-γ level in the large dose group was lower than that in the small dose group(p<0.05).(4)ImmunohistochemistryThe type III collagen staining was brownish yellow,mainly concentrated in the interstitial lung and peribronchial area,and the expression level of type III collagen was significantly higher in the remaining groups compared with the blank control group(p<0.01);compared with the model group,the expression level of type III collagen was significantly lower in the high,medium and low doses of Bushentongluo formula,and pirfenidone group(p<0.01);compared with the low dose of Bushentongluo formula,the expression level of type III collagen was significantly higher in the high The expression level of type III collagen was significantly higher in the Pirfenidone group compared with the Bushentongluo formula low dose group(p<0.01).The LN staining was brownish yellow and was expressed in the bronchial wall,alveolar lumen and alveolar interstitium.Compared with the blank control group,the LN expression in the model group and pirfenidone group was significantly higher than that in the blank control group(p<0.01),while there was no significant difference between the rest of the groups and the blank control group;compared with the model group,the LN expression level was significantly lower in the high,medium and low dose groups of Bushentongluo Formula(p<0.01).The LN expression levels in the high,medium and low dose groups of Bushentongluo Formula were significantly lower than those in the pirfenidone group(p<0.01).(5)Western blotComparison between groups of p-JAK2/JAK2:compared with the blank control group,the model group and pirfenidone group,the p-JAK2/JAK2 levels were significantly increased in the high and medium dose groups of Bushentongluo Formula(p<0.01);compared with the model group,the p-JAK2/JAK2 levels were significantly decreased in the medium dose group of Bushentongluo Formula(p<0.01),compared with the medium dose group of Bushentongluo Formula.Pirfenidone group,p-JAK2/JAK2 levels were increased in the high and low dose groups of Bushentongluo Formula(p<0.05).Comparison between p-STAT3/STAT3 groups:compared with the blank control group,the p-STAT3/STAT3 levels were significantly higher in the model group,the high,medium and low dose groups of Bushentongluo Formula and the pirfenidone group(p<0.01),and significantly lower in the medium dose group of Bushentongluo Formula compared with the model group(p<0.05),while the levels of p-STAT3/STAT3 were significantly lower in the rest of the groups.There was no significant difference between the remaining groups.Conclusion1 Clinical part(1)The high prevalence of RA-ILD is in older women.(2)In RA-ILD,deficiency of lung and kidney qi and paralysis of the lung and lung complexes are common,suggesting that deficiency of lung and kidney qi and paralysis of the lung and lung complexes may be the most common TCM evidence of RA-ILD,and the pathogenesis of the disease can be summarized as "deficiency,stasis and phlegm".(3)The CRP and ESR levels tend to increase in cases of lung-kidney qi deficiency and lung-loop paralysis,suggesting that the degree of inflammation may be higher in cases of RA-ILD with lung-kidney qi deficiency and lung-loop paralysis.2 Experimental part(1)The formula can reduce the degree of arthritis in RA-ILD rats.(2)Bushentongluo Formula can reduce pulmonary fibrosis in RA-ILD rats.The mechanism may be related to increasing the release of IFN-γ,inhibiting JAK2/STAT3 signaling pathway and reducing extracellular matrix deposition. |
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