Inhibition of genistein against LPS-induced proinflammatory response in microglia
10.3969/j.issn.1674-8115.2019.05.002
- Author:
Hong-Mei WANG
1
Author Information
1. Department of Neurology, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University
- Publication Type:Journal Article
- Keywords:
Genistein;
Inflammation;
Lipopolysaccharide (LPS);
Microglia;
NLRP3 inflammasome;
Reactive oxygen species (ROS)
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2019;39(5):446-450
- CountryChina
- Language:Chinese
-
Abstract:
Objective • To investigate the effect of genistein (Gen) on lipopolysaccharide (LPS)-induced proinflammatory response in microglia. Methods • Primary microglia were isolated from C57BL/6J mice 1 d after birth, which were divided into 3 groups, i.e. control group, LPS group, and Gen+LPS group. Microglia in LPS group and Gen+LPS group were incubated with LPS (1 μg/mL) for 24 h, and the cells in Gen+LPS group were also pretreated with Gen (10 μmol/L) for 0.5 h. The expression of CD11b was measured by Western blotting analysis. Besides, mitochondrial and intracellular reactive oxygen species (ROS) levels were monitored by MitoSOXTM Red and CM-H2DCFDA staining, respectively. NLRP3 (NLR family pyrin domain containing 3) inflammasom was detected by immunofluorescence. Caspase-1 activity and interleukin-1β (IL-1β) levels were evaluated by colorimetric assay kit and ELISA kit, respectively. Results • Compared with control group, LPS increased CD11b protein expression, and mitochondrial and intracellular ROS levels in microglia (P<0.05). And Gen obviously reversed LPS-induced mitochondrial and intracellular ROS accumulation as well as reduced CD11b expression (P<0.05). In addition, LPS enhanced fluorescence intensity of NLRP3 inflammasome, caspase-1 activity and IL-1β secretion in microglia when compared with control group (P<0.05), while Gen significantly attenuated these effects (P<0.05). Conclusion·Gen can inhibit LPS-induced proinflammatory response including mitochondrial ROS accumulation, activation of NLRP3 inflammasome, and IL-1β secretion in microglia.
