| Objective:1.Based on the outpatient medical records of Professor Xiaoyan Gong in treating interstitial lung disease(ILD),this paper summarizes Professor Xiaoyan Gong’s thinking and experience in treating ILD and guides the clinical treatment of ILD.2.Using metabonomics as the research method,we explored the changes in the serum metabolite spectrum of mice with bleomycin-induced pulmonary fibrosis treated by Xiankefang,searched for the potential biomarkers of pulmonary fibrosis and the internal mechanism of Xiankefang’s intervention in pulmonary fibrosis,and laid a theoretical foundation for clinical research on the prevention and treatment of ILD by traditional Chinese medicine.Methods:1.Clinical research: Collect and sort out the ILD outpatients who Professor Xiaoyan Gong treated in Jilin Provincial Hospital of Traditional Chinese Medicine from August 2018 to July 2021,screen the cases according to the inclusion and exclusion criteria,establish an Excel data database,and enter the standardized data one by one,including basic information,symptoms,tongue and pulse,syndrome types and prescriptions,and carry out frequency analysis,association rule analysis and cluster analysis based on the traditional Chinese medicine inheritance computing platform(V2.5).2.Experimental study: The mouse model of pulmonary fibrosis was prepared by bleomycin intratracheal infusion.Observe the general condition,lung coefficient,and Masson’s staining of mice,and evaluate the effect of Xiankefang on the pathological morphology of lung tissue in mice with pulmonary fibrosis;Detection of TGF-β in serum and HYP in lung tissue by ELISA,detection of α-SMA in lung tissue by immunohistochemistry,confirming the anti-fibrosis effect of Xiankefang;Ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS)technology was used to detect the serum samples of mice with pulmonary fibrosis.At the same time,Principal Component Analysis(PCA),Partial least squares Discriminant Analysis(PLS-DA),Orthogonal Partial least squares Discriminant Analysis(OPLS-DA)multivariate statistical analyses were performed on the data.The potential differential metabolites of the disease were screened by fold change(FC ≥ 1.2 or FC ≤ 0.833)combined with Variable important in projection(VIP>1),P<0.05.Determine the ability of potential biomarkers to predict disease through ROC curve analysis;Through KEGG Pathway,we analyzed the metabolic pathway that Xiankefang interfered with the significant enrichment of serum differential metabolites in mice with pulmonary fibrosis and expounded its mechanism of action.Result:1.Clinical research(1)Baseline results: ILD patients were more male than female;The age of onset is mainly middle-aged and elderly.(2)Frequency analysis results: The main clinical symptoms were cough,dyspnea with rapid and short breath,and expectoration;The distribution of syndrome types is mainly the syndrome of lung and kidney deficiency and vein obstruction,the syndrome of heat toxin blocking the lung and the syndrome of qi deficiency and blood stasis;The drugs are mainly classified as cough and resolving phlegm medicines,tonic medicines,antipyretic,rheumatism medicines,relieving drugs,and blood-activating medicines;The nature of herb medicine is relatively peaceful,with almost as many cold herbs as warm herbs;The smell of the herbs is primarily sweet,bitter,and spicy and belongs to the lung meridian mostly.(3)Results of association rules: Obtained a combination of three herbs commonly used in clinical practice,namely "polygonum cuspidatum,clematis,Siegesbeckia orientalis," "astragalus,ligusticum chuanxiong,and earthworm," and some herb-medicine pair,namely "aster and coltsfoot," "white front and radix peucedani," "poria cocos and fried atractylodes macrocephaly," In addition,astragalus,polygonum cuspidatum,earthworm,aster,coltsfoot,and angelica sinensis are at the center of the association network,and are core drugs for the treatment of ILD.(4)Complex system entropy clustering results: Twenty groups of three herbs and 1group of four were obtained.Their functions involve warming the lung and relieving the exterior,clearing heat and detoxifying,replenishing qi and unblocking collaterals,nourishing yin and promoting fluid production,reducing cough and resolving phlegm,soothing the liver and regulating qi,and warming the kidney and strengthening the spleen;Five new prescriptions of traditional Chinese medicine were obtained,mainly for “warming the lung,filling qi and clearing collaterals,” and “nourishing yin,replenishing qi and promoting fluid production.”2.Experimental research(1)Effect of Xiankefang on the pathological morphology of lung tissue in mice with pulmonary fibrosis: Body weight and lung coefficient: Compared with the model group,the weight in the pirfenidone group and Xiankefang group increased significantly(p<0.01).In contrast,the lung coefficient decreased(p<0.05).Masson staining: the alveolar structure in the lung tissue of the model group mice was damaged,and a large number of collagen fibers were observed;Compared with the model group,the fibrosis degree of mice in the pirfenidone group and the Xiankefang group was significantly reduced(p<0.05).(2)Study on anti-fibrosis of Xiankefang: Compared with the blank control group,the content of TGF-β,α-SMA,and HYP of mice in each pulmonary fibrosis group increased significantly(p<0.05);Compared with the model group,the content of TGF-β,α-SMA,and HYP of mice in the pirfenidone group and the Xiankefang group were significantly decreased(p<0.01),and there was no significant difference between the pirfenidone group and Xianke group.(3)Serum metabolomics of pulmonary fibrosis mice and the intervention mechanism of Xiankefang: There were 71 serum differential metabolites between the blank group and the pulmonary fibrosis model group,and 15 potential biomarkers,including lipids,amino acids,nucleotides,and other small molecular metabolites,were obtained through Pub Chem and HMDB’s biological function query and literature search.The ROC curve analysis accepted lysophosphatidylcholine,3-methyl indole,and tryptophan AUC>0.95.The main metabolic pathways related to pulmonary fibrosis are white matter digestion and absorption,aminoacyl t RNA biosynthesis,and tryptophan metabolism.Xiankefang mainly affects protein digestion and absorption,biosynthesis of aminoacyl t RNA,arginine and proline metabolism,and tryptophan metabolism to interfere with forming lung fibers by regulating metabolites such as glutamic acid,lysine,L-asparagine,methionine,phenylalanine,and proline.Conclusion:1.Clinical researchBased on the analysis of data mining results of the included cases,the clinical experience and medication rule of Professor Xiaoyan Gong in the treatment of ILD is summarized as follows:(1)The location of viscera disease: The diseased viscera is in the lung,and the disease site is in the lung collaterals.It is closely related to the liver,spleen,and kidney and even affects the heart in the later stage.(2)Summary of pathogenesis: The pathogenesis factors mainly involve evil,deficiency,phlegm,blood stasis,and toxin,which are based on lack,marked by phlegm,blood stasis,and toxin,and disturbed by evil(poison).The etiology and pathogenesis are deficiency of lung,deficiency of qi,and blood stasis.The critical pathogenesis is obstruction of lung collaterals.(3)Syndrome differentiation and classification: Professor Xiaoyan Gong advocates the system of syndrome differentiation and treatment of "three phases and six types," namely,the acute attack period: the syndrome of cold-fluid in the lung with evil(wind),and the syndrome of hot phlegm blocking lung entrapment;Chronic sub-acute: stagnation of liver and spleen,stagnation of qi and blood stasis;Severe recurrent period: the syndrome of lung and kidney deficiency and vein obstruction,and syndrome of yang qi deficiency and water overflowing the heart and lung.(4)Characteristics of differentiation and treatment: The law of medication reflects the treating separately for different pathogenic factors,using warm herbs,reinforcing insufficiency and reducing excessiveness,and focusing on dredging the collaterals.(5)Drug use experience:(1)Drugs are used according to the stages of the disease: in the acute attack stage,the method of clearing the lung and removing arthralgia is always used;The treatment of Chronic sub-acute is to replenish qi and dredge collaterals;In severe recurrent period,it is essential to warm the spleen and kidney.(2)Combination of disease differentiation and syndrome differentiation: Regard "Polygonum cuspidatum,Clematis,Siegesbeckia orientalis" as the targeted drug for treating "Feibi," then combine with traditional syndrome differentiation,and emphasize the complexity and comprehensiveness of the disease.2.Experimental research(1)Xianke Fang significantly improved the pathological morphology of lung tissue in mice with pulmonary fibrosis.(2)Xiankefang may reduce TGF-β of promote-fibrosis cytokine that inhibit the transformation of fibroblasts into myofibroblasts,reduce collagen deposition,and inhibit the occurrence and development of lung fibers.(3)Potential biomarkers related to pulmonary fibrosis are involved in maintaining cell structure and function,regulation of fibrosis-related cytokines,oxidative stress,inflammation,and related chemokine regulation.It is speculated that inflammatory damage and repair disorders,oxidative stress,and fibroblast proliferation and transformation are essential mechanisms leading to pulmonary fibrosis;Among them,the abnormality of amino acid metabolism is most closely related to the occurrence and development of PF;LPC,tryptophan,3-MI or become biomarkers for disease diagnosis.Xiankefang mainly inhibits pulmonary fibrosis by regulating amino acid metabolism,maintaining cell function,repairing inflammatory damage,and controlling oxidative balance. |
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