RESUMO
ObjectiveTo investigate the effects of Aconiti Coreani Radix and Typhonii Rhizoma on the urinary metabolites of gerbils with stroke by non-targeted metabolomics technique， and then to clarify the mechanism of the two， as well as their similarities and differences. MethodTwenty-four gerbils were randomly divided into control group（CG）， model group（MG）， Aconiti Coreani Radix group（RA） and Typhonii Rhizoma group（RT）. Except for the CG， ischemic stroke model was constructed using right unilateral ligation of gerbil carotid artery in the remaining groups. Except for the CG and MG， rats in the other groups received whole powder suspension（0.586 mg·g-1） was administered for 14 days. The neurological deficit in each group was scored by Longa scoring on days 0， 3， 7 and 14. After the end of administration， the serum， brain tissue and urine of gerbils in each group were collected， and the rate of cerebral infarction was detected by 2，3，5-triphenyltetrazolium chloride（TTC）， and the levels of interleukin（IL）-6， tumor necrosis factor（TNF）-α， malondialdehyde（MDA）， superoxide dismutase（SOD）， glutathione（GSH）， and nitric oxide（NO） in serum and brain tissue were determined by enzyme-linked immunosorbent assay（ELISA）. The urine metabolomics of gerbils in each group was studied by ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Orbitrap-MS）， and the data were processed by multivariate statistical analysis， and differential metabolites were screened based on value of variable importance in the projection（VIP） of the first principal component>1 and t-test P<0.05. Metabolic pathway analysis of the screened differential metabolites was performed using Kyoto Encyclopedia of Genes and Genomes（KEGG） database and Metaboanalyst 5.0. ResultCompared with the CG， the neurological deficit score was significantly increased in the MG（P<0.05）， compared with the MG， the neurological deficit scores in the RA and RT were significantly reduced after 7 d and 14 d（P<0.05）. Compared with the CG， the rate of cerebral infarction was significantly increased in the MG（P<0.05）， compared with the MG， the rates of cerebral infarction in the RA and RT were significantly reduced（P<0.05）. Compared with the CG， the levels of IL-6， TNF-α， and MDA in the serum and brain tissue of gerbils from the MG were significantly increased（P<0.05）， and the levels of SOD， GSH and NO were significantly reduced（P<0.05）. Compared with the MG， Aconiti Coreani Radix and Typhonii Rhizoma could down-regulate the levels of IL-6， TNF-α and MDA， and up-regulated the levels of SOD， GSH and NO. A total of 112 endogenous differential metabolites were screened by urine metabolomics， of which 16 and 26 metabolites were called back by Aconiti Coreani Radix and Typhonii Rhizoma， and could be used as potential biomarkers for both treatments in stroke gerbils， respectively. The results of the pathway analysis showed that both Aconiti Coreani Radix and Typhonii Rhizoma had regulatory effects on arginine and proline metabolism， pyrimidine metabolism， and aminoacyl-tRNA biosynthesis. In addition， Aconiti Coreani Radix could also regulate riboflavin metabolism， Typhonii Rhizoma could also regulate purine metabolism， glycine， serine and threonine metabolism， arachidonic acid metabolism， biosynthesis of pantothenate and coenzyme A， and β-alanine metabolism. ConclusionBoth Aconiti Coreani Radix and Typhonii Rhizoma have better therapeutic effects on stroke， with Aconiti Coreani Radix having stronger effects. From the metabolomics results， the main metabolic pathways regulated by Aconiti Coreani Radix involve amino acid metabolism， oxidative stress and so on， while Typhonii Rhizoma mainly involve amino acid metabolism， lipid metabolism， energy metabolism， etc.
RESUMO
Objective:To analyze the association of the genetic variations of rs2383206 and rs2383207 in 9p21region with the coronary heart disease (CHD) in the Chinese Han population,and to explore whether chromosome 9p21 is a susceptibility region for CHD.Methods:Case-control study was conducted.A total of 580 CHD patients were selected as case group,and 539 cases of non-cardiovascular disease patients or normal people with matched age and sex were selected as control group.The rs2383206 and rs2383207 loci of the subjects were genotyped with Sequenom MassARRAY time of flight mass spectrometer (TOF).Results:The smoking,waist-to-hip ratio (WHR),hypertension,diabetes mellitus,systolic blood pressure (SBP),diastolic blood pressure (DBP) and total cholesterol (TC) of the subjects in two groups were statistically different (P＜0.05).Compared with control group,the ratios of patients with smoking,hypertension and diabetes mellitus of the patients in case group were increased (P＜0.05);the WHR,SBP,DBP and TC level were also increased (P＜0.05).There was no significant difference in the genotypic distribution of rs2383206 between case group and control group (x2 =4.623,P＞0.05),while the genotypic distribution of rs2383207 was statistically different (x2 =8.936,P＜0.05);the distribution frequency of AA genotype in case group (8.3％) was significantly lower than that in control group (13.6％) (P＜0.05).Conclusion:Smoking,WHR,hypertension,diabetes mellitus,SBP,DBP and TC may be the risk factors for CHD;the AA genotype of 9p21 rs2383207 loci may be the protective genotype of CHD.
RESUMO
Mogroside V is the most abundant [approximately 0.50%] cucurbitane-type triterpene glycoside in Siraitia grosvenorii and exhibits significant antitussive, expectorant, anti-carcinogenic, and anti-inflammatory effects. A sensitive, robust and selective liquid chromatography tandem with mass spectrometry [LC-MS/MS] was developed and validated for the determination and pharmacokinetic investigation of mogroside V in rat plasma. Samples were prepared through an one-step deproteinization procedure with 250 microL of methanol to a 75-microL plasma sample. Plasma samples were effectively separated on a Shiseido Capcell Pak UG120 C18 column [2.0 × 50mm, 3.0microm] using a mobile phase consisting of methanol: water [60:40, v/v] with an isocratic elution program. The running time for each sample was 7.0 min and the elution times of mogroside V and IS were 2.0 and 4.8 min, respectively. The detection relied on a triplequadrupole tandem with mass spectrometer equipped with negative-ion electrospray ionization interface by selectedreaction monitoring [SRM] of the transitions at m/z 1285.6 - 1123.7 for mogroside V and m/z 1089.6 - 649.6 for IS. The calibration curve was linear over the range of 96.0-96000 ng/mL with a limit of quantitation [LOQ] of 96.0ng/mL. Intra-day and inter-day precisions were both <10.1%. Mean recovery and matrix effect of mogroside V in plasma were in the range of 91.3-95.7% and 98.2-105.0%, respectively. This method was successfully applied in the pharmacokinetic study of mogroside V after intravenous or intraperitoneal administration of 1.12mg/kg mogroside V in rats
RESUMO
Objective To investigate effect of high glucose on the function of endothelial and the underlying mechanisms in human umbilical vascular endothelial cells (HUVECs).Methods The experiment was divided into 4 groups: normal group (NG), low dose group (LG), middle dose group (MG) and high dose group ( HG) .The concentration of glucose in the culture medium was 5.5, 10, 20, 30 mmol/L in the 4 groups, respectively.The HUVECs was cultured for 0, 24, 48 h.At different time point, the cell viability were measured by MTT.The secretary content of nitric oxide (NO) in the supernatant were detected using test kit.The extraction of protein were extracted for Western blot analysis to detect the expression of endothelial nitric oxide synthase (eNOS).ResuIts Compared with normal group at same time point (cultured for 24 h), the cell viability and the content of NO were significantly decreased in LG, MG(P<0.05).The expression of eNOS in HG were markedly reduced (P<0.01).Compared with normal group at same time point (cultured for 48 h), the cell viability decreased significantly in HG (P <0.05).The expression of eNOS were markedly decreased 11.91, 25.72 and 34.50% in LG, MG and HG, respectively.A rising trend of cell viability were found in NG, LG and MG, but a descending trend were found in HG within 48 h.ConcIusion The cell viability were significantly affected by high glucose.The endothelial dysfunction induced by high glucose may be associated with the reduction of eNOS and NO production.