Korean J Physiol Pharmacol.  2015 Nov;19(6):491-497. 10.4196/kjpp.2015.19.6.491.

Epigallocatechin-3-Gallate (EGCG) Attenuates Traumatic Brain Injury by Inhibition of Edema Formation and Oxidative Stress

• Zhang B
• Wang B
• Cao S
• Wang Y

• KMID: 2070788
• DOI: http://doi.org/10.4196/kjpp.2015.19.6.491

Abstract

Traumatic brain injury (TBI) is a major cause of mortality and long-term disability, which can decrease quality of life. In spite of numerous studies suggesting that Epigallocatechin-3-gallate (EGCG) has been used as a therapeutic agent for a broad range of disorders, the effect of EGCG on TBI remains unknown. In this study, a weight drop model was established to evaluate the therapeutic potential of EGCG on TBI. Rats were administered with 100 mg/kg EGCG or PBS intraperitoneally. At different times following trauma, rats were sacrificed for analysis. It was found that EGCG (100 mg/kg, i.p.) treatment significantly reduced brain water content and vascular permeability at 12, 24, 48, 72 hour after TBI. Real-time PCR results revealed that EGCG inhibited TBI-induced IL-1beta and TNF-alpha mRNA expression. Importantly, CD68 mRNA expression decreasing in the brain suggested that EGCG inhibited microglia activation. Western blotting and immunohistochemistry results showed that administering of EGCG significantly inhibited the levels of aquaporin-4 (AQP4) and glial fibrillary acidic protein (GFAP) expression. TBI-induced oxidative stress was remarkably impaired by EGCG treatment, which elevated the activities of SOD and GSH-PX. Conversely, EGCG significantly reduced the contents of MDA after TBI. In addition, EGCG decreased TBI-induced NADPH oxidase activation through inhibition of p47phox translocation from cytoplasm to plasma membrane. These data demonstrate that EGCG treatment may be an effective therapeutic strategy for TBI and the underlying mechanism involves inhibition of oxidative stress.

Keyword

Anti-oxidant; Edema; EGCG; NADPH; Traumatic brain injury

MeSH Terms

Animals
Blotting, Western
Brain
Brain Injuries*
Capillary Permeability
Cell Membrane
Cytoplasm
Edema*
Glial Fibrillary Acidic Protein
Immunohistochemistry
Microglia
Mortality
NADP
NADPH Oxidase
Oxidative Stress*
Quality of Life
Rats
Real-Time Polymerase Chain Reaction
RNA, Messenger
Tumor Necrosis Factor-alpha
Water
Glial Fibrillary Acidic Protein
NADP
NADPH Oxidase
RNA, Messenger
Tumor Necrosis Factor-alpha
Water

Figure

• 

  Fig. 1 EGCG reduced TBI-induced brain tissue water content and vascular permeability. After TBI, rats were administered with PBS or 100 mg/kg EGCG intraperitoneally and sacrificed at indicated time. (A) Brain tissue water content was analyzed and the percentage of tissue water content was calculated as (WW-DW)/WW×100%. (B) Evans blue dye (0.2 ml/100 g) was injected through the femoral vein. The brain tissue was homogenized in formamide and the supernatant was quantitated by spectrophotometric analysis at 620 nm. Data are shown as means±S.E.M. #p<0.05 for sham group versus TBI group, *p<0.05 for TBI group versus EGCG treatment group (n=5).

• 

  Fig. 2 EGCG ameliorated TBI-induced inflammation. Quantitative real-time PCR analysis of the levels of expression of proinflammatory genes including (A) CD68, (B) IL-1β and (C) TNF-α after TBI. Data are shown as means±S.E.M. #p<0.05 for sham group versus TBI group, *p<0.05 for TBI group versus EGCG treatment group (n=5).

• 

  Fig. 3 EGCG inhibited TBI-induced AQP4 and GFAP expression in brain tissue. Western blot analysis for AQP4 and GFAP protein levels at 12, 24, 48 and 72 h after TBI. Expressions of GAPDH were shown as loading controls. Bands were analyzed by densitometry. Quantitative data were shown. Data are shown as means±S.E.M. #p<0.05 for sham group versus TBI group, *p<0.05 for TBI group versus EGCG treatment group.

• 

  Fig. 4 Immunohistochemistry analysis for AQP4 and GFAP positive cells at 24 and 72 hour after TBI. The DAB-positive areas in 5 regions were measured using microscope and quantitative analysis of the immunoreactivity was performed. Data are shown as means±S.E.M. #p<0.05 for sham group versus TBI group, *p<0.05 for TBI group versus EGCG treatment group.

• 

  Fig. 5 EGCG increased SOD and GSH-PX activities and decreased MDA activity. After TBI, the left brain hemispheres of mice were homogenized and the supernatant was used for the measurement of SOD (A), GSH-PX (B) and MDA activity (C). Data are shown as means±S.E.M. #p<0.05 for sham group versus TBI group, *p<0.05 for TBI group versus EGCG treatment group (n=5).

• 

  Fig. 6 EGCG inhibited TBI-induced p47phox translocation from cytoplasm to plasma membrane. Translocation of the subunits of NADPH oxidase was analyzed by Western blot at 24 h after TBI. Expressions of GAPDH were shown as loading controls for cytoplasm. Expressions of gp91phox were shown as loading controls for membrane. Bands were analyzed by densitometry. Quantitative data were shown. Data are shown as means±S.E.M. #p<0.05 for sham group versus TBI group, *p<0.05 for TBI group versus EGCG treatment group (n=5).

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