Effect And Mechanism Research Of Panax Notoginseng Saponins R1 On The Repair Of Asthma Airway Epithelial Damage By Glucocorticoids

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Effect of Panax notoginseng saponins R1 on Glucocorticoid Inhibition of Airway Epithelial Damage Repair in Asthmatic MiceOBJECTIVE: To investigate whether panax notoginseng saponins R1 has a therapeutic effect on glucocorticoid-induced injury repair of airway epithelium in asthmatic mice.METHODS: A mouse model of asthma was established by using house dust mites(HDM),and glucocorticoid-dexamethasone(Dex)was intraperitoneally injected half an hour before the last 3 shots,while intraperitoneal injection of panax notoginseng saponins R1(PNS-R1)solution.The lung function of the mice was detected by invasive pulmonary function tracheal intubation within 24 hours after the last challenge.The mouse bronchoalveolar lavage fluid(BALF)specimens were collected,and the total number of cells in the alveolar lavage fluid was counted by a cell counter.The cell classification was counted after staining and the number of cells shed in BALF.The trachea and left lung tissues of the mice were collected for dehydrated paraffin-embedded sections.HE staining and immunohistochemical E-cadherin staining were performed to observe the damage of airway epithelial cells in mice.The apoptosis of airway epithelial cells was observed by TUNEL staining.The expression of caspase3 in airway epithelium was detected by immunohistochemistry and Western blot.The secretion of mucus in airway was observed by PAS glycogen staining.The content of epithelial damage-related factors in alveolar lavage fluid was detected by ELISA.RESULTS: 1.The lung ventilation resistance,airway inflammation and airway mucus secretion of asthmatic mice were significantly increased compared with the control group.Both Dex and PNS-R1 could significantly reduce lung ventilation resistance,airway inflammation and airway mucus hypersecretion in asthmatic mice,but Dex and Lung ventilation resistance,airway inflammation,and airway mucus secretion in the PNS-R1 co-treatment group did not differ from those treated with Dex alone.2.Dex treatment of asthmatic mice significantly increased the number of epithelial cells and IL-33 and TSLP in the BALF,and decreased the integrity of airway epithelial cells.Adding PNS-R1 to Dex would significantly reduce the shedding of BALF.The number of epithelial cells and the damage-related factors IL-33 and TSLP increased the integrity of airway epithelial cells.3.Dex treatment of asthmatic mice significantly increased the percentage of airway epithelial cell apoptosis and caspase3 protein expression.Adding PNS-R1 to Dex would significantly reduce the percentage of airway epithelial cell apoptosis and caspase3 protein expression.CONCLUSION:Dex can significantly inhibit the repair of airway epithelial cells in asthmatic mice and promote the apoptosis of airway epithelial cells.The simultaneous use of PNS-R1 in Dex-treated mice can significantly improve the repair of airway epithelial damage in Dex-suppressed asthmatic mice.The role of both,while reducing the apoptosis of airway epithelial cells,and ultimately promote the repair of airway epithelial damage in asthmatic mice.Part ? Effect of Panax notoginseng saponins R1 on Glucocorticoids Inhibiting Growth of Bronchial Epithelial CellsOBJECTIVE: To observe whether the panax notoginseng saponins R1 can improve the effect of glucocorticoids on the growth of bronchial epithelial cells.METHODS: Human bronchial epithelial cell line(16HBE)was cultured in vitro,and cells were intervened with Dex and PNS-R1.Cell growth was detected by CCK-8 technique;Transwell assay and scratch assay were used to detect the migration ability of bronchial epithelial cells;cell cloning experiments and cell cycle assays were used to detect the proliferation of bronchial epithelial cells.At the same time,TUNEL experiment and flow-type Annexin-V technique were used to detect the apoptosis of bronchial epithelial cells.Western blot was used to detect the expression of apoptosis protein caspase3 in bronchial epithelial cells.RESULTS: 1.Dex significantly inhibited the proliferation and migration of 16 HBE.The migration and proliferation of 16 HBE cells were significantly increased after PNS-R1 intervention,and there was no difference between the two groups.2.Dex can promote the apoptosis of 16 HBE cells.After the intervention of PNS-R1,the apoptosis of 16 HBE cells was significantly reduced,which was no difference with the control group.3.The expression of apoptotic protein caspase3 was significantly increased in cells treated with Dex.The apoptosis protein caspase3 was significantly lower in the cells treated with PNS-R1 than in the Dex group.The difference was statistically significant.CONCLUSION: Dex can significantly promote the migration and proliferation of bronchial epithelial cells and promote the apoptosis of bronchial epithelial cells.The simultaneous intervention of PNS-R1 in 16 HBE cells treated with Dex can significantly improve the inhibitory effect of Dex on the migration and proliferation of bronchial epithelial cells.Dex-induced apoptosis of bronchial epithelial cells ultimately promotes the growth of bronchial epithelium.Part ? Effect of Panax notoginseng saponins R1 on the repair of airway epithelial damage in asthma by glucocorticoids through the influence of GR? on MAPK signaling pathwayOBJECTIVE: To explore the specific mechanism of panax notoginseng saponins R1 to improve the inhibition of glucocorticoids on airway epithelial damage in asthma.METHODS: Peripheral blood samples from children with asthma and control group were collected and the expression of GR? protein in monocytes was detected by Western blot.Western blot was used to detect the changes of glucocorticoid receptor(GR,Glucocorticoid receptor)and MAPK signaling pathway in lung tissue samples of Dex or/and PNS-R1 mice.Lentiviral transfection knockdown of GR? gene in asthmatic mice,collecting bronchoalveolar lavage fluid samples from mice,and counting the number of cells detached from BALF was counted.The tracheal and left lung tissues of the mice were collected for dehydrated paraffin-embedded sections.HE staining and immunohistochemical E-cadherin staining were performed to observe the damage of airway epithelial cells in mice.The ELISA was used to detect the content of epithelial damage-related factors in alveolar lavage fluid.The expression of GR? was detected by Western blot.16 HBE was cultured in vitro,and the expression of GR and MAPK signaling pathways in cells treated with Dex or/and PNS-R1 were detected by Western blot,immunohistochemistry and co-immunoprecipitation.Then use the si-RNA knockdown the GR? in the cells and then intervene with the cells with Dex or/and PNS-R1;Transwell test and scratch test to detect bronchial epithelium cell migration ability;cell cloning experiments and cell cycle assays to detect changes in bronchial epithelial cell proliferation.Western blot analysis was used to detect the expression of MAPK signaling pathway.RESULTS: 1.The expression of GR? protein in peripheral blood samples of patients with clinical asthma using glucocorticoids was significantly lower than that in patients without asthma using glucocorticoids.The expression of asthma in the glucocorticoid group was also lower than that in the control group,and the difference was statistically significant.2.Dex can significantly inhibit the expression of GR? in 16 HBE cells,reduce the binding of GR?/GR?,and significantly reduce the activation of MAPK signaling pathway.After the addition of PNS-R,the expression of GR? increased,which promoted the binding of GR?/GR?,which promoted the activation of MAPK signaling pathway.3.Knockdown of GR? in 16 HBE cells,PNS-R1 improved the ability of Dex to inhibit the migration and proliferation of 16 HBE cells,and the activation of MAPK signaling pathway was significantly reduced.4.Dex can significantly inhibit the expression of GR? in lung tissue and airway epithelial cells of asthmatic mice and reduce the activation of MAPK signaling pathway.The expression of GR? increased after the intervention of PNS-R1,which promoted the activation of MAPK signaling pathway.5.PNS-R1 improved the ability of Dex to inhibit airway injury repair in asthmatic mice after knocking down GR? in asthmatic mice.CONCLUSION:Dex may inhibit the activation of MAPK signaling pathway and inhibit the repair of airway epithelial cells in asthma by inhibiting the expression of GR?,reducing the negative regulation of GR?.PNS-R1 may increase the activation of MAPK signaling pathway by reducing the inhibitory effect of Dex on GR?,thereby improving the role of Dex in inhibiting the repair of airway epithelial damage in asthma.