OBJECTIVETo investigate the effect of respiratory syncytial virus (RSV)-related pulmonary infection on endogenous metabolites in large intestinal mucosa in BALB/c mice using metabolomics technology based on gas chromatography-mass spectrometry (GC-MS).

METHODSMice were randomly divided into a control group and a RSV pneumonia model group (n=16 each). The mouse model of RSV pneumonia was established using intranasal RSV infection (100×TCID, 50 μL/mouse, once a day). After 7 days of intranasal RSV infection, the mice were sacrificed and GC-MS was used to identify endogenous metabolites and measure the changes in their relative content in colon tissue. SMCA-P12.0 software was used to perform principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) for endogenous metabolites in colon tissue. The differentially expressed metabolites in colon tissue were imported into the metabolic pathway platform Metaboanalyst to analyze related metabolic pathways.

RESULTSPCA and OPLS-DA showed significant differences between the control and RSV pneumonia model groups. A total of 32 metabolites were identified in the colon tissue of the mice with RSV pneumonia. The RSV pneumonia model group had significant increases in the content of leucine, isoleucine, glycine, alanine, arachidonic acid, and lactic acid, which were related to the valine, leucine, isoleucine, arachidonic acid, and pyruvic acid metabolic pathways.

CONCLUSIONSRSV pneumonia might cause metabolic disorders in the large intestinal tissue in mice.

Amino Acids, Branched-Chain ; metabolism ; Animals ; Female ; Gas Chromatography-Mass Spectrometry ; Intestinal Mucosa ; metabolism ; Intestine, Large ; metabolism ; pathology ; Lung ; pathology ; Mice ; Mice, Inbred BALB C ; Pneumonia, Viral ; metabolism ; Respiratory Syncytial Virus Infections ; metabolism

OBJECTIVE To develop a sensitive and selective ultra performance liquid chromatography-triple quadrupole mass spectrometer(UPLC-MS/MS) method for quantification of eight components(chrysin,baicalein,apigenin,orohylin A, wogonin,baicalin,wogonoside,oroxin A) in Schisandra chinensis Georgi.METHODS The separation was performed on a Thermo BDS Hypersil C1 8(2.1 mm×100 mm,2.3 μm) column.The mobile consisted of acetonitrile (A) and 0.1% formic acid aqueous with a gradient elution at a flow-rate of 0.25 mL/min.The detection was acquired by MRM.The column temper-ature was 40 ℃.RESULTS The linear ranges of chrysin,baicalein,apigenin,orohylin A,wogonin,baicalin,wogonoside, oroxin A were 0.975~62.5 ng/mL(r 2 =0.999 8),4.875~2 500 ng/mL(r 2 =0.999 7),0.975~62.5 ng/mL(r 2 =0.999 4), 0.097 5~12.5 ng/mL(r 2 =0.999 6),0.048 83~6.25 ng/mL(r 2 =0.999 9),4.875~2 500 ng/mL(r 2 =0.999 8),0.488 3~250 ng/mL(r 2 =0.999 9),0.488 3 ~ 250 ng/mL(r 2 = 0.999 8),respectively.The recoveries were 98.75%,96.44%, 101.13%,99.41%,102.26%,98.45%,96.75%,99.42%.CONCLUSION The proposed method enables quantification for the study of the compounds of chrysin,baicalein,apigenin,orohylin A,wogonin,baicalin,wogonoside,oroxin A in Schisan-dra chinensis Georgi.

OBJECTIVE To detect the relative abundance changes of metabolites in urine and feces of BALB/c mice to seek the biomarkers that contribute to the diagnosis by GC-MS technology.METHODS BALB/c mice were challenged intranasally to establish the RSV pneumonia models.Urine and feces were collected respectively,oximated,derivatized and detected by GC-MS.RESULTS 38 metabolites in urine were detected,among which lactic acid,propanoic acid and L-proline were statistical-ly significant(P<0.05).Moreover,43 metabolites in feces were detected,and leucine,aspartic acid,cystine,tyrosine and pyrimidine had statistical significance(P<0.05).CONCLUSION GC-MS technology is feasible for seeking biomarkers in u-rine and feces of mice infected with RSV pneumonia.

Tripterygium wilfordii Hook. f. induced-hepatotoxicity was the main limitation for its usage in clinic. Qingluo Tongbi formulation showed obvious attenuation for hepatotoxicity in clinic and fundamental research in vivo. To explore the potential mechanism of the attenuation, we conducted a study on the plasma metabolomic profiles of T. wilfordii and Qingluo Tongbi formulation in rats by a sensitive gas chromatography-mass spectrometry (GC-MS/MS) method. In plasma samples, a total of 72 compounds were analyzed by EI source MS, and were successfully identified by matching NIST database. The semi-quantification results were then calculated by OPLS-DA model with SIMCA-P 13.0 software. The three groups were clearly distinguished in OPLS-DA score plot. In addition, the observation values of Qingluo Tongbi formulation showed the obvious trend towards the control levels, suggesting the detoxicity effect of the formulation. Variation metabolites were further analyzed by VIP and One Way ANOVAs, and the results showed a significant increase in compounds of glycogenic amino acids, such as alanine, proline, serine and glutamine after the administration of T. wilfordii, indicated that the tissue proteins were decomposed and amino acids were leakage into blood. Qingluo Tongbi formulation could reverse the amino acids into normal level. On the contrary, the levels of glucose, lactic acid and hydroxy butyrate decrease, and the formulation can relieve the disorder in the levels of lactic acid, suggesting the regulation of the energy metabolism. Additionally, the level of branched chain amino acid was decreased, suggested the toxicity was induced, but the formulation cannot increase it into the normal levels. Nevertheless, all the above results suggested that the classical Qingluo Tongbi formulation displayed the liver protection effect by adjusting the amino acid levels and regulating the energy metabolism. Qingluo Tongbi formulation was developed based on traditional Chinese medicine theory "detoxicity compatibility", and contained Panax notoginseng (Burk.) F. H. Chen to nourish blood and absorb clots. Modern pharmacology suggested that its liver protection effect was correlated with the promotion of protein synthesis. Another important herb is Rehmannia glutinosa Libosch., which can regulate the energy metabolism. Both were consistent with the metabolomic results in this study, which explained the potential mechanism of "detoxicity compatibility" theory. Therefore, the currently developed metabolomic approach and the obtained results would be highly useful for the comprehensive toxicity studies for other herbal medicines and various complex deoxicity formulations.