calpain and epigallocatechin-gallate

The impact of prophylactic supplementation of N-acetyl-cysteine (NAC) and epigallocatechin gallate (EGCG) on intramuscular expression of proteolytic genes after unaccustomed eccentric muscle contractions was investigated.. Thirty apparently healthy males (mean ± SD: 20.0 ± 1.8 years, 175 ± 7.1cm, 76.1 ± 16.9 kg) ingested daily either 1,800 mg of NAC or 1,800 mg of EGCG (98% total polyphenols, 80% total catechins, and 50% EGCG), or 1,000 mg of a glucomannan placebo (PLA) in a double blind, prophylactic fashion for 14 days. Subjects then completed an unaccustomed eccentric exercise bout (100 repetitions at 30 °s(-1)) using the dominant knee extensors. Skeletal muscle biopsies were collected from the vastus lateralis at baseline and both 6 and 24h after exercise. The expression of proteolytic genes [i.e., muscle ring-finger 1 (MuRF1), atrogin-1, α-type 20S subunit C2 (HC2), α-type 20S subunit C3 (HC3), ubiquitin protein ligase 3B (UBE3B), μ-calpain, and m-calpain] was quantified using real-time RT-PCR. Separate 3 × 3 (group × time) repeated measures ANOVAs were used to analyze changes in gene expression over time between groups.. No significant group × time interactions were detected between groups for the expression of any of the atrogenes or calpains (p>0.05). Significant main effects for time identified increases in MuRF1 (6h: 5.3 ± 10.8 fold; p=0.046), UBE3B (6h: 5.3 ± 7.7 fold; p=0.006; 24h: 3.3 ± 4.5 fold; p=0.005), and m-calpain expression (6h: 2.7 ± 4.4 fold; p=0.045) in all participants following exercise. Increases approached significance in HC2 (6h: 1.9 ± 2.4 fold; p=0.079; 24h: 1.6 ± 1.9 fold; p=0.084) and m-calpain expression (24h: 1.8 ± 2.3 fold; p=0.084) following exercise.. Prophylactic supplementation of NAC and EGCG did not impact acute changes in skeletal muscle proteolytic gene expression following eccentric exercise. Eccentric muscle contractions elevated MuRF1 and UBE3B, while m-calpain and HC2 mRNA tended to increase.

Topics: Acetylcysteine; Calpain; Catechin; Dietary Supplements; Double-Blind Method; Exercise; Gene Expression; Humans; Male; Muscle Contraction; Muscle Proteins; Muscle, Skeletal; Nontherapeutic Human Experimentation; SKP Cullin F-Box Protein Ligases; Tripartite Motif Proteins; Ubiquitin-Protein Ligases; Young Adult

It has been discovered that proteasome inhibitors are able to induce tumor growth arrest or cell death and that tea consumption is correlated with cancer prevention. Here, we show that ester bond-containing tea polyphenols, such as (-)-epigallocatechin-3-gallate (EGCG), potently and specifically inhibit the chymotrypsin-like activity of the proteasome in vitro (IC(50) = 86-194 nm) and in vivo (1-10 microm) at the concentrations found in the serum of green tea drinkers. Atomic orbital energy analyses and high performance liquid chromatography suggest that the carbon of the polyphenol ester bond is essential for targeting, thereby inhibiting the proteasome in cancer cells. This inhibition of the proteasome by EGCG in several tumor and transformed cell lines results in the accumulation of two natural proteasome substrates, p27(Kip1) and IkappaB-alpha, an inhibitor of transcription factor NF-kappaB, followed by growth arrest in the G(1) phase of the cell cycle. Furthermore, compared with their simian virus-transformed counterpart, the parental normal human fibroblasts were much more resistant to EGCG-induced p27(Kip1) protein accumulation and G(1) arrest. Our study suggests that the proteasome is a cancer-related molecular target of tea polyphenols and that inhibition of the proteasome activity by ester bond-containing polyphenols may contribute to the cancer-preventative effect of tea.

Topics: Calpain; Caspase 3; Caspases; Catechin; Cell Cycle Proteins; Cell Line, Transformed; Cyclin-Dependent Kinase Inhibitor p27; Cyclin-Dependent Kinases; Cysteine Endopeptidases; Cysteine Proteinase Inhibitors; DNA-Binding Proteins; Esters; Fibroblasts; Flavonoids; Humans; I-kappa B Proteins; Jurkat Cells; Male; Microtubule-Associated Proteins; Models, Molecular; Molecular Conformation; Multienzyme Complexes; NF-KappaB Inhibitor alpha; Phenols; Polymers; Prostatic Neoplasms; Proteasome Endopeptidase Complex; Structure-Activity Relationship; Tea; Tumor Cells, Cultured; Tumor Suppressor Proteins