| Objective: To explore the etiology and pathogenesis of fibrotic interstitial lung disease(F-ILD)by data mining of TCM syndrome types and clinical medication.To explore the mechanism of TCM intervention in pulmonary fibrosis through animal experiments.In order to provide new ideas for the treatment of pulmonary fibrosis.Methods:Part Ⅰ: Collected 97 cases of F-ILD patients from October 2019 to October 2021 in The Affiliated Hospital of Shandong University of Traditional Chinese Medicine Professor Zhang Wei outpatient clinic,using Microsoft Excel 2019 for TCM syndrome type frequency statistics,ancient and modern medical case cloud platform for clinical drug data mining,analysis of drug use rules,summarize the etiology and pathogenesis.Part Ⅱ: 1.48 male SD rats were divided into 6 groups by random number table method:blank group,model group,pirfenidone group,astragalus-salvia miltiorrhiza low-dose group,Astragalus-salvia miltiorrhiza medium-dose group and Astragalus-salvia miltiorrhiza high-dose group.Except blank group,the animal model of pulmonary fibrosis was induced by bleomycin dripping into trachea,and the animal model was sacrificed after28 days of intragastric intervention.2.HE staining and Masson staining were used to observe the pathological changes of lung tissues.3.Determination of hydroxyproline content in rat lung tissue by alkaline hydrolysation;4.Protein expression levels of PI3 K,p-PI3 K,AKT,p-Akt,m TOR,p-MTOR,LC3,Beclin1 and P62 in rat lung tissues were detected by Western blotting.5.LC3 level was detected by immunofluorescence.Results:Part Ⅰ: a total of 97 cases,175 prescriptions,73 herbs were included.The frequency statistics of syndrome types showed that the majority of F-ILD was deficiency syndrome,and the syndrome types containing qi deficiency and blood stasis accounted for 45.36% of the total.High frequency drugs were astragalus membranaceus,Epimedium herba,Poria cocos,Chinese yam,cornus officinalis and salvia miltiorrhiza.Astragalus membranaceus was the most frequently used drug.The four main qi are flat,cold,warm,mild and mild cold,the five tastes are sweet,bitter and bitter,and the meridian of return is lung,kidney and liver.The main types of drugs were qi tonifying drugs,phlegm-reducing drugs,relieving surface drugs,relieving cough and antiasthmatic drugs,promoting blood circulation and removing stasis drugs.As the frequency of astragalus membranaceus was the highest,it was found that astragalus membranaceus was mostly associated with promoting blood circulation,invigorating qi and reinforcing Yang,and the most associated medicinal smells with promoting blood circulation and removing blood stasis were salvia miltiorrhiza,peony bark,Angelica sinensis and Ligusticum chuanxiong.Astragalus-Salvia miltiorrhiza is the strongest drug pair associated with qi and blood circulation.Four groups of drugs were obtained by cluster analysis,the second group mainly used to remove blood stasis and dissolve phlegm,relieve cough and relieve asthma,the second group mainly used to nourish qi and promote blood circulation,warm Yang and nourish blood.Two core prescriptions were obtained by complex network analysis,which were mainly for cleansing and supplementing.Part Ⅱ:1.HE staining,Masson staining and hydroxyproline staining: except for the blank group,inflammatory cell infiltration,collagen deposition and pulmonary fibrosis changes of different degrees were observed in the other groups,which proved that the modeling was successful.The changes of tissue structure destruction,inflammation and fibrosis in pirfenidone group and astragalus salvia miltiorrhiza dose groups were less than those in model group,and the improvement degree of astragalus salvia miltiorrhiza dose group was dose-dependent.Analysis of hydroxyproline content confirmed the trend of pathological changes.2.Observe the activation of PI3K/AKT/m TOR signaling pathway in each group:Western blotting method: Compared with blank group,the expression of P-PI3K/PI3 K,P-Akt /AKT,p-m TOR /m TOR in model group increased(p<0.01).Compared with model group,the expressions of P-PI3 K /PI3 K,P-Akt /AKT and P-m TOR /m TOR in pirfenidone group and Astragalus salvia miltiorus-dose group were decreased(p<0.01),astragalus and Salvia miltiorrhiza were dose-dependent(p<0.05 or p<0.01).3.Observe the autophagy level of rats in each group:Western blotting method: Compared with blank group,Beclin1,LC3-Ⅱ/LC3-Ⅰexpress ion in model group decreased(p<0.01).P62 expression increased(p<0.01).Comp ared with model group,the expressions of LC3-Ⅱ/LC3-Ⅰand Beclin1 in pirfenidone group and astragalus salviae salviae groups were increased(p < 0.01 or p < 0.05),p62 expression decreased(p<0.01).Beclin1 expression was increased in the medium and high dose group compared with the low dose group(p<0.01),there was no s ignificant difference between Beclin1 medium-high dose groups(p<0.05).The expr ession of LC3-Ⅱ/LC3-Ⅰincreased in a dose-dependent manner,and the expression of P62 decreased in a dose-dependent manner(p<0.01).Immunofluorescence method: a large number of LC3 fluorescence spots were observed in the lung tissues of rats in the blank group,while only a very small number of LC3 fluorescence spots were observed in the lung tissues of rats in the model group.Compared with model group,the number of spots in pirfenidone group and Astragalus-salvia miltiorrhiza dose groups increased in different degrees.Conclusion:1.F-ILD is a group of this deficiency standard disease,clinical pathogenesis is attributed to positive deficiency,stasis,phlegm phlegm,phlegm stagnation,after the end of the lung and kidney damage,qi deficiency and blood stasis is an important syndrome type,treatment to benefit qi and promote blood circulation,qi and phlegm,lung and kidney,fuzhengquxie as the principle.Professor Zhang Wei attached great importance to the application of qi and blood activating drugs in clinical practice,among which Astragalus membranaceus-Salvia miltiorrhizae was the strongest drug associated with qi and blood activating.At the same time,phlegm-reducing,qi regulating and surface relieving treatments were carried out along with the syndrome.Use of mild,small effect macro,multi-purpose flat,sweet products,rarely use major cold,hot products.2.The drug pair of "Astragalus-Salvia miltiorrhiza" can improve and inhibit pulmonary fibrosis to a certain extent,which may be related to reducing the phosp horylation levels of PI3 K,AKT and m TOR and activating autophagy. |
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