| Objective: This study used the Curcumae Rhizoma and Panax Notoginseng with the function of stasis eliminating,which is derived from Yifei Sanjie prescription,a clinically effective prescription of Professor Li Qingsheng,a famous Chinese medicine practitioner in Yunnan Province,in order to observe the effect and mechanism of the main active components of Curcumae Rhizoma and Panax Notoginseng,including Curcumol,Curcumin and Notoginseng Triterpenes,on the pulmonary fibrosis in mice,and to explore whether it can alleviate pulmonary fibrosis.Then,it was observed whether Curcumol,Curcumin,Notoginseng triterpenes could enhance the autophagy level of lung tissue to interfere with pulmonary fibrosis by inhibiting the PI3K/AKT/m TOR signaling pathway,which will provide a new idea for clinical prevention and treatment of the disease.Method: 1.Replication of mouse pulmonary fibrosis model and grouping: The mouse pulmonary fibrosis model was replicated by bleomycin atomization once a day for a total of 7 days.The experiment was divided into control group,model group,Yisan component(part of Yifei Sanjie prescription)intervention group,pirfenidone group and inhibitor group.2.Preparation of drugs and drug intervention: according to the proportion of Curcumol,Curcumin and Notoginseng Triterpenes in Yifei Sanjie prescription,prepare the premixed liquid of some components of Yifei Sanjie Prescription,and give the premixed liquid of some components of Yifei Sanjie Prescription to the intervention group and inhibitor group of Yifei Sanjie Prescription for gavage administration;The pirfenidone group was given pirfenidone by gavage administration.The inhibitor group was treated with premixed liquid and chloroquine.The control group and the model group were fed with the same amount of sterile water.After successful replication of the model,gavage administration once a day for 28 consecutive days.3.Index observation: The pathological changes of lung tissue in mice were observed by HE and Masson staining.Lung hydroxyproline was used to detect collagen content in lung tissue.The relative expression levels of TGF-β1,α-SMA and Collagen-Ⅲm RNA were detected by Q-PCR.The number of autophagosomes was observed under electron microscope.The expression levels of autophagy-related proteins LC3-II and Beclin1 and the expression levels of PI3K/AKT/m TOR signaling pathways p-PI3 K,pAKT and p-m TOR proteins were detected by Western blot.Results: 1.Curcumin,curcumol,and Panax notoginseng saponins alleviate bleomycin-induced pulmonary fibrosis in mice 1.1 lung tissue pathological section results In the model group,the alveolar septum was significantly thickened,and some alveolar wall rupture and alveolar fusion were observed.Inflammatory cell infiltration,vascular wall,flaky or flaky collagen fibers were observed around the bronchial wall,and collagen fibers were observed in the alveolar space.In the intervention group,the alveolar septal thickening was observed,the alveolar wall rupture and fusion were improved,and the deposition of tracheal,blood vessel and interstitial collagen fibers was significantly reduced.1.2 Collagen test results The relative expressions of hydroxyproline and Collagen-III m RNA in the lung tissue of the model group were significantly increased(P<0.05).Compared with the model group,after 28 days of administration,the Yishen component intervention group The relative expression of hydroxyproline and Collagen-III m RNA in lung tissue decreased compared with the model group,and the difference was statistically significant(P<0.05).1.3 Relative expression levels of fibrosis-related factors TGF-β1 and α-SMA m RNA The relative expression of TGF-β1 and α-SMA m RNA in the lung tissue of the model group was significantly increased(P<0.05).Compared with the model group,TGF-β1 in the lung tissue of the intervention group.The relative expression of α-SMA m RNA decreased,and the difference was statistically significant(P<0.05).Compared with the inhibitor group,the relative expression of TGF-β1 and α-SMA m RNA in the lung tissue of the Yisan component intervention group was different.The difference was statistically significant(P < 0.05).2.Curcumin,curcumol,and Panax notoginseng saponins enhance the autophagy level of lung tissue cells 2.1 electron microscopy observation of autophagosome results The number of autophagosomes in the lung tissue of the model group was significantly reduced.Compared with the model group,the number of autophagosomes in the lung tissue of the mice in the Yisan component intervention group increased significantly.2.2 Expression of autophagy-related proteins LC3-II and Beclin1 The expressions of autophagy-related proteins LC3B-II and Beclin1 in the lung tissue of the model group were significantly lower than those in the control group(P<0.05).Compared with the model group,the Yishen component intervention group was used in the lung tissue of mice.The expression of LC3B-II and Beclin1 was increased,and the difference was statistically significant(P<0.05).3.Chloroquine inhibits the expression of fibrosis-related factors after autophagy After inhibition of autophagy,compared with the Yishen component intervention group,the inflammatory infiltration was significantly increased in the components of the Yifei Sanjie prescription + inhibitor group,the alveolar septum was widened,and some alveolar wall rupture and alveolar fusion,trachea and lung were observed.The deposition of collagen fibers around the interstitial was significantly increased.The relative expression of hydroxyproline and Collagen-3 m RNA in the lung tissue of some components of Yifei Sanjie Prescription + inhibitor group was increased(P<0.05).The relative expression of TGF-β1 and α-SMA m RNA in lung tissue of Yifei Sanjie Prescription+Inhibitor group was significantly increased(P<0.05).4.Curcumol,Curcumin and Notoginseng Triterpenes inhibit the regulation of autophagy by PI3K/AKT/m TOR signaling pathway The expression of p-PI3 K,p-AKT and p-m TOR protein in the lung tissue of the model group was significantly higher than that in the control group(P<0.05).Compared with the model group,the intervention group was small.The expression of p-PI3 K,pAKT and p-m TOR protein in the lung tissue of rats was decreased,the difference was statistically significant(P<0.05).At the same time,the number of autophagosomes in the lung tissue of mice in the intervention group was significant.The content of LC3 BII and Beclin1 protein increased,and the difference was statistically significant(P<0.05).Conclusion: 1.The expression of fibrosis-related factors TGF-β1,α-SMA and Collagen-III decreased by the partial components of Yifei Sanjie Prescription,Curcumol,Curcumin and Notoginseng Triterpenes,which reduced the deposition of extracellular matrix and collagen fibers.It relieves bleomycin-induced pulmonary fibrosis in mice.2.The components of Yifei Sanjie Prescription,Curcumol,Curcumin and Notoginseng Triterpenes promote the expression of autophagy-related proteins LC3B-II and Beclin1,inhibiting the PI3K/AKT/m TOR signaling pathway,possibly alleviating pulmonary fibrosis is one of the important signal transduction mechanisms. |
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