| Airway reconstruction, also called airway remodeling, is one of the basic pathological changes of bronchial asthma attacks for a long time repeatedly, the main performance is the airway wall thickening, and the result is a continuous airway hyperresponsiveness. Many inflammatory products (TGF-β1 or IL-17) in lung tissue of asthma which influence airway wall collagen deposition and promote airway sub-epithelial fibrosis, ultimately lead to airway remodeling as the wide range of histopathological structural changes. The purpose of the current study was to investigate the effect of panax notoginseng saponins (PNS) which possess properties viz. anti-inflammation, promoting blood circulation to remove blood stasis, and immunity enhancement activities to prevent airway remodeling in mouse model of bronchial asthma.Therefore, considering notoginseng pharmacological effects and molecular mechanism of the airway reconstruction process, we would have explored the effect of notoginseng total saponin on bronchial asthma airway reconstructionin mice and its molecular mechanism. Methods:Eighty Kunming female mice were selected and randomly divided into Panax notoginseng saponins treatment (High and Low PNS) group, Dex treatment group, positive control group (asthma group) and control group. Each group was fed eight mice, and there were 10 groups. Mice were sensitized by intraperitoneal (i.p.) injections of OVA-AL(OH)3 suspension and exposed to 2% OVA solution for challenge after sensitization.0.5h before challenge on alternate days for 6 weeks to induce long-term (LTE) protocols,and short-term exposure (STE) protocol exposed consecutively for 2 weeks. After the completion of modeling, the cardiopulmonary tissues were removed for tissue perfusion. The left lung was removed for staining by hematoxylin and eosin (H&E) to assess airway pathological alteration under a light microscopy (the ratio of airway stenosis, the ratio of erosion and necrosis of airway epithelial cells, the ratio of goblet cell metaplasia, the ratio of inflammatory cell infiltration and the ratio of smooth muscle hyperplasia). The levels of TGF-β1 and collagen Ⅲ were detected by immunohistochemical staining, and analyzed using a Leica imaging svstem and computerized image analysis software to semi-quantified. The levels of IL-17 in right lung tissue supernatants were measured by ELISA. Differences in protein were using iTRAQ proteomics technology analysis, and using bioinformatics data retrieval technology, including Gene Ontology analysis (GO, Gene Ontology), and protein in general statistical analysis, protein and protein interactions and KEGG (kyoko Encyclopedia of Genes and Genomes) pathway analysis. Results:①After cardiopulmonary perfusion, the treatment groups have different levels of red blood silk, drops of blood and the pink. Positive control group in the lung tissue surface was presented a chunk of passive congestion, clots and dark red, while the lavage of negative control group is clean, showing a white, clear lines of lung tissue.② Under a microscope in H&E staining, bronchial asthma group was surrounded by a large number of inflammatory cell infiltration, airway structure severely was damaged, drug treatment group have different degrees of improvement, and the negative control group have no significant pathological symptoms; ③The expression of TGF-β1and collagen Ⅲ in treatment group was significantly lower than that of the asthma group (P<0.001). ④The thickness of airway basement membrane of long term in asthma group, PNS group and control group were (0.7893±0.014)、(0.7216±0.016)、(0.5655±0.012)μm respectively, which was significantly higher in the asthma group than those of the PNS group and control group (P<0.05). ⑤The level of IL-17 in long term of high PNS group decreased less to a half of asthma group (P>0.05). No significant difference was found compared with the control group (p=0.94, p=0.23). Proteomics research results show that in which to get the basic information of the overall analysis of five groups of proteins, including the number of total protein of identification for 79572, the mass distribution of protein (focused on 10-160kDa), isoelectric point distribution (protein concentration in PI=5～7, the amount is 820), distribution of the length of peptides (the distribution of the most widely for 4～6 amino acids, is 13482) and the number of peptides (the amount is 36308. the number of peptides was concentrated upon 10 and proteins number decreased along with the number of matching peptides increasing), distribution of protein peptides’ sequence coverage (the largest protein proportion is 33% when the peptides sequence coverage is in 0～5%, the amount is 799) distribution of protein abundance ratio, and statistical figure of each group differences in protein quantity. We select asthma-LTE vs control-LTE (distinct protein of up-regulation is 84, and down-regulate is 57) and High PNS-LTE vs asthma-LTE (distinct protein of up-regulation is 83, and down-regulate is 170) to carry out bioinformatics analysis. GO functional annotations in protein analysis, protein gene identification to its Molecular Function, location of the cell (Cellular Component), and participation in Biological processes of the proportion in its various components all have different degrees of difference. Through a string of protein interaction maps can tell the difference between the two groups to compare protein quantity and category, and also the different kinds of potential interaction networks. KEGG differences in protein pathway enrichment analysis can be seen higher protein and lower protein different effect which in the pathway of asthma-LTE vs control-LTE and high PNS-LTE vs asthma-LTE. Interested to find the different proteins including actinin, Iqgapl, Arpc2, Decorin, Atplal and GC-s closely associated with asthma airway formation process, and according to their passages, it can be presumably known the notoginseng intervention effect on asthma. The experimental results show that notoginseng total saponin can effectively improve airway reconstruction of bronchial asthma in mice, and inhibit the airway inflammatory, at the same time through the analysis of the differences of notoginseng asthma produce proteins rise or fall, can provides the reference for the molecular mechanism of asthma of panax notoginseng. |
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