Abstract
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.
Original language | English (US) |
---|---|
Pages (from-to) | 32-39 |
Number of pages | 8 |
Journal | Biochemical and Biophysical Research Communications |
Volume | 411 |
Issue number | 1 |
DOIs | |
State | Published - Jul 22 2011 |
Externally published | Yes |
Bibliographical note
Funding Information:This work was supported by the National High-tech R&D Program of China (863 Program) ( 2009AA022703 , 2009AA022704 ), National Natural Science Foundation of China ( 30873089 ), Program for Changjiang Scholars and Innovative Research Team in University ( IRT0946 ), Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province, Open Foundation of Innovative Platform in University of Hunan Province of China ( 10K078 ), and the Science and Technology Plan Key Grant of Hunan Province of China ( 2009TP4068-2 ).
Keywords
- HO-1
- Magnesium lithospermate B
- Nrf2
- Reative oxygen species