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Qu J.,Central South University | Ren X.,Shanghai Green Valley Pharmaceutical Co. | Hou R.-Y.,Central South University | Dai X.-P.,Central South University | And 6 more authors.
Biochemical and Biophysical Research Communications | Year: 2011

Objectives: To investigate the effects of magnesium lithospermate B (LAB) on intracellular reactive oxygen species (ROS) production induced by high dose of glucose or H2O2, we explored the influences of LAB on the expression of heme oxygenase-1 (HO-1) and nuclear factor E2-related factor-2 (Nrf2) in HEK293T cells after treatment with high dose of glucose. Materials and methods: The total nuclear proteins in HEK293T cells were extracted with Cytoplasmic Protein Extraction Kit. The ROS level was determined by flow cytometry. The mRNA and protein expression of HO-1 and Nrf2 were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot. Results: LAB reduced the ROS production in HEK293T cells cultured under oxidative stress. High dose of glucose enhanced the expression of HO-1 mRNA and HO-1 protein in a time-dependent manner. LAB enhanced the expression of HO-1 mRNA and HO-1 protein in a dose-dependent manner treated with high dose of glucose. The amount of Nrf2 translocation was enhanced after cells were pretreated with 50μmol/L or 100μmol/L LAB. Silencing of Nrf2 gene eliminated the enhanced expression of HO-1 protein induced by high dose of glucose plus LAB. Conclusions: LAB plays an important role against glucose-induced intracellular oxidative damage. The enhanced expression of HO-1 mRNA and HO-1 protein caused by LAB is regulated via Nrf2 signal pathway. © 2011 .

Sheng F.,Central South University | Ren X.,Shanghai Green Valley Pharmaceutical Co. | Dai X.,Central South University | Xu X.,Central South University | And 6 more authors.
Journal of Central South University (Medical Sciences) | Year: 2011

Objective To investigate the effect of nicotinamide mononucleotide ( NMN ) on insulin secretion and gene expressions of pancreatic and duodenal homeobox 1 ( PDX-1 ) and forkhead box-containing protein 0-1 ( Fox01 ) , which were important transcription factors for insulin secretion. Methods Insulin secretion level in RIN-m5f cells was detected by rat insulin ELISA detection kit. The mRNA expression levels of PDX-1 and FoxOI in RIN-m5f cells were analyzed by real-time PCR. The protein expression of PDX-1 was measured by Western blot. Results Insulin secretion levels in RIN-m5f cells treated with repaglinide (10 nmol/L) plus NMN (100 μmol/L) was significantly higher than those in the blank control, the DMSO control group, and the NMN ( 50 μmol/L) treated group (P<0.05). The mRNA expression levels of PDX-1 in RIN-m5f cells treated with NMN ( 10, 50 and 100 μmol/L) for 36 h were significantly higher than those in the control group (P<0.05, P<0.01, and P<0.001, respectively). There was marked differences in the mRNA expression levels of PDX-1 among different concentrations of NMN (P <0.001 ) , but no significant differences in the mRNA expression level of Fox01 (P > 0.05). No significant difference was found in the protein expression levels of PDX-1 in RIN-m5f cells treated by NMN (50, 100, and 200 (μmol/L) for 36 or 48 h compared with the control group (P >0.05). Conclusion NMN can stimulate insulin secretion and upregulate the mRNA expression of PDX-1 in RIN-m5f cells.

Qin C.-Z.,Central South University | Ren X.,Shanghai Green Valley Pharmaceutical Co. | Tan Z.-R.,Central South University | Chen Y.,Central South University | And 5 more authors.
Biomedical Chromatography | Year: 2014

A sensitive and high-throughput inhibition screening liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous quantification of five probe metabolites (7-hydroxycoumarin, CYP2A6; 4-hydroxytolbutamide, CYP2C9; 4′-hydroxymephenytoin, CYP2C19; α-hydroxymetoprolol, CYP2D6; and 1-hydroxymidazolam, CYP3A4) for in vitro cytochrome P450 activity determination in human liver microsome and recombinant. All the metabolites and the internal standard, tramadol, were separated on a Waters 2695 series liquid chromatograph with a Phenomenex Luna C18 column (150×2.0mm, 5μm). Quality control samples and a positive control CYP inhibitor were included in the method. The IC50 values determined for typical CYP inhibitors were reproducible and in agreement with the literature. The method was selective and showed good accuracy (99.13-103.37%), and inter-day (RSD<6.20%) and intra-day (RSD<6.13%) precision. Also, the incubation extracts of the sample were stable at room temperature (20°C) for 48h and for 96h in the autosampler (4°C). The presented method is the first HPLC-MS/MS method of this combination for simultaneous detection of the five metabolites 7-hydroxycoumarin, 4-hydroxytolbutamide, 4′-hydroxymephenytoin, α-hydroxymetoprolol and 1-hydroxymidazolam in a single-run process. It is possible that the high-quality and -throughput cocktail provides suitable information in drug discovery and screening for new drug entities. © 2013 John Wiley & Sons, Ltd.

Qin C.-Z.,Central South University | Ren X.,Shanghai Green Valley Pharmaceutical Co. | Zhou H.-H.,Central South University | Mao X.-Y.,Central South University | Liu Z.-Q.,Central South University
International Journal of Clinical and Experimental Medicine | Year: 2015

Salvianolic acid B (Sal B), which is purified from Danshen, is a popular herb extract. Sal B has anti-oxidative, anti-inflammatory, anti-hypoxic, anti-arteriosclerotic and anti-apoptotic properties. This substance can also ameliorate brain injury or neurodegenerative diseases. The listed drug Salvianolate, which contains a substantial amount of Sal B, has been used for the treatment of coronary heart disease. Our present work aimed to evaluate the inhibitory effect of salvianolate on seven cytochrome P450 isoforms (CYP450), namely, CYP1A2, CYP2A6, CYP2E1, CYP2C9, CYP2C19, CYP2D6 and CYP3A4, in human liver microsomes (HLMs) and recombinant enzymes through high-performance liquid chromatography (HPLC) assay. Salvianolate have a potent inhibitory effect on CYP3A4 activity with IC50 values of 1.438 (HLMs) and 3.582 (recombinant cDNA-expressed CYP3A4) mg/L, respectively. Salvianolate strongly dose, but not time-dependently decreased CYP3A4 activity in HLMs. The typical Lineweaver-Burk plots showed that Salvianolate inhibited CYP3A4 activity noncompetitively, with a Ki value of 2.27 mg/L in HLMs. Other CYP450 isoforms are not markedly affected by Salvianolate. These findings indicate that salvianolate may be involved in potential drug interactions when co-administrated with CYP3A4 substrates. © 2015, E-Century Publishing Corporation. All rights reserved.

Shi R.-B.,Beijing University of Chinese Medicine | Wang Y.-Y.,Chinese Academy of Sciences | Lv S.-T.,Shanghai Green Valley Pharmaceutical Co.
Zhongguo Zhongyao Zazhi | Year: 2015

According to the coorelative analyses on Chinese medicine essence, dosage forms and quality control level, it expounds the precise concept of Chinese medicine, and its quality advantages and characteristics in this paper, furthermore discusses how to achieve the ideal drug and Chinese medicine equality precision in expectation. Base on the Chinese medicine essence, using the concept of nature medicine and its drug system to construct Chinese medicine effective material basis and its drug, with the correlative analyses of whole view and reductionism, the problems of uncertainty quality of original natural medicinal resource and preparation may well be solved, and further with the macroscopic to microcosmic contruction of drug sytem, the precision in expectation of Chinese medicin equality and higher production lever may well be achieved.

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