ABSTRACT
UHPLC-Q-TOF/MS metabonomics technology was used to clarify the metabolic regulation pathways by which Platycodon total saponins (PTS) exert antitussive and expectorant effects in a mouse cough model, in which coughing is induced by concentrated ammonia, and in a phenol red excretion model. After approval by the Experimental Animal Ethics Committee of Jiangxi University of Chinese Medicine (Approval No. JZLLSC-20190235), the mice were randomly divided into a normal group, a model group, a positive drug group and a PTS group. Endogenous metabolites in mouse serum were identified by UHPLC-Q-TOF/MS. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for multivariate analysis. Metabolic pathways were analyzed by the Metaboanalyst platform. The results show that PTS can significantly prolong the cough latent period and cough frequency of mice, and significantly increase phenol red excretion. UHPLC-Q-TOF/MS identified 19 metabolites related to cough, and PTS significantly decreased 16 of them; 17 metabolites related to expectoration were identified, and PTS decreased the levels of all. Metabolic pathway analysis showed that linoleic acid metabolism, arachidonic acid metabolism and glycerophospholipid metabolism were the main pathways involved in serum metabolite changes in this mouse cough model. Linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, phenylalanine metabolism and α-linolenic acid metabolism were the main pathways involved in serum metabolite changes in the phenol red excretion model. This study is the first to elucidate the regulation of antitussive and expectorant metabolic pathways and the effect of PTS on these pathways.
ABSTRACT
Objective: To screen the differentially expressed proteins of saponins in Pulsatillae Radix inhibiting the proliferation and induce apoptosis on NCI-H460 tumor cells based on proteome technology using nano LC-LTQ-Orbitrap-MS/MS, and preliminarily speculate the potential mechanism. Method: NCI-H460, SK-OV-3 and SGC-7901 tumor cells were cultured in vitro. Methylthiazoletetrazolium (MTT) assay was used to detect the inhibitory rate of saponins in Pulsatillae Radix on three tumor cell lines. Effect of saponins in Pulsatillae Radix on apoptosis was analyzed by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining flow cytometry and 4',6-diamidino-2-phenylindole (DAPI) staining. Apoptosis was analyzed using flow cytometry and DAPI stain. Nano LC-LTQ-Orbitrap-MS/MS was used to investigate the changes in the protein profiles on NCI-H460 cells treated with saponins in Pulsatillae Radix. Proteins exhibiting differential expression were analyzed by DAVID Bioinformatics Resources 6.8 and Kyoto encyclopedia of genes and genomes (KEGG) database. The differentially expressed proteins were verified by Western blot. Result: Saponins in Pulsatillae Radix could inhibit the proliferation of NCI-H460, SK-OV-3 and SGC-7901 tumor cells and induce apoptosis of NCI-H460 tumor cells. Effect of Saponins in Pulsatillae Radix on the proliferation and apoptosis of NCI-H460 tumor cells was mainly related to the regulation of biological function of ribosome, glycolysis/gluconeogenesis and other biological processes. It was possible to induce apoptosis of NCI-H460 tumor cells by interfering mitogen-activated protein kinase (MAPK) signaling pathway and regulating the Caspase pathway. Conclusion: Saponins in Pulsatillae Radix can inhibit the proliferation and induce the apoptosis of NCI-H460 tumor cells, the mechanism may be related to the intervention of MAPK signaling pathway and the regulation of Caspase pathway. These findings are helpful to elucidate the molecular mechanism of the anti-tumor effect of saponins in Pulsatillae Radix.
ABSTRACT
In this study, we investigated the protective effect of the antioxidant N-acetyl-L-cysteine (NAC) on the lung inflammation caused by ozone (O) exposure in mice. Thirty-two C57BL/6 mice were randomly divided into control group, Ogroup, O+NAC group and NAC group. Mice were exposed to O(1.0 ppm) or fresh air for 3 h on the day 1, day 3 and day 5, respectively. NAC (100 mg/kg) was intraperitoneally applied to the mice 1 h before each exposure. At 24 h after the 3-time exposure, the alveolar wall structure was severely damaged and the infiltrated inflammatory cells were apparent perivascularly and peribronchiolarly. Significant increases in the total white blood cell count, macrophage, lymphocyte and neutrophil counts, as well as total protein concentration were observed in the bronchoalveolar lavage fluid (BALF) (P < 0.05). The IL-6, IL-8 (P < 0.01) and MDA levels (P < 0.05) in the lung homogenates were elevated coherently. Administration of NAC could attenuate the alveolar wall structure damage induced by Oexposure and reduce the amount of infiltrated inflammatory cells, total and differential leukocyte counts (P < 0.05), as well as the IL-6, IL-8 (P < 0.01) and MDA release (P < 0.05). Western blotting results showed that the Oexposure up-regulated the p38 MAPK and NF-κB p65 protein expression in the lung tissue of mice (P < 0.05), which could be alleviated by NAC (P < 0.05). These results indicated that NAC could protect against O-induced pulmonary inflammation in mice. The beneficial effect of NAC might be related with the p38 MAPK and NF-κB p65 signal pathway.
Subject(s)
Animals , Mice , Acetylcysteine , Antioxidants , Bronchoalveolar Lavage Fluid , Interleukin-6 , Lung , Mice, Inbred C57BL , NF-kappa B , Neutrophils , Ozone , PneumoniaABSTRACT
<p><b>OBJECTIVE</b>To observe the symptoms of inattention, hyperactivity among obstructive sleep apnea hypopnea syndrome (OSAHS) children, also to investigate the effects of surgery (tonsillectomy and adenoidectomy or adenoidectomy alone) on the changes of sleep architecture and inattention-hyperactivity score (IHS).</p><p><b>METHODS</b>Between June 2004 and may 2007, eighty children diagnosed as OSAHS with overnight polysomnography (PSG) were included in this study, only sixteen children had complete pre-op and post-op PSG data. Thirty children with vocal cord nodules were selected as control group. DSM-IV-derived IHS was evaluated by neurologist. All OSAHS children accepted surgery (tonsillectomy and adenoidectomy or adenoidectomy alone) and IHS evaluation. The pre-op and post-op sleep architecture and IHS were compared with that of control group.</p><p><b>RESULTS</b>(1) The median IHS 80 OSAHS children was higher than that it in control group (0.89 vs 0.17) and the difference was significant (Z = -4. 276, P < 0.05). After surgery, it showed a significant reduction in IHS (0.44 vs 0.89, t = 6.219, P < 0.05). (2) Twenty-five OSAHS children had pre-op IHS greater than 1.25 and nine had post-op IHS greater than 1.25, while only three children in control group had IHS greater than 1.25. The difference was statistically significant (chi2 = 5.192, 9.56 respectively, P < 0.05). (3) For sixteen OSAHS children who had both pre-op and post-op PSG data, a decrease in the percentage of phase 1 sleep and an increase in the percentage of phase 2 sleep, slow wave sleep (SWS) and rapid eye movement (REM) sleep were observed in six months after surgery and the difference was significant (t = 12.2, -5.4, -6.3, - 8.1 respectively, P < 0.001). After surgery, apnea-hypopnea index (AHI) decreased from 13.9 times/h to 1.5 times/h while lowest saturation of blood oxygen (LSaO2) increased from 0.855 to 0.940 (t = 5.3, - 3.7 respectively, P < 0.01). REM sleep percentage and LSaO2 was still lower than that of control group six months after surgery.</p><p><b>CONCLUSIONS</b>Children with OSAHS showed significantly impaired attention and hyperactivity as compared with control group. Improvement of behavior and sleep architecture were observed after adenoidectomy and tonsillectomy.</p>