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Partial cloning,tissue distribution and effects of epigallocatechin gallate on hepatic 3-hydroxy-3-methylglutaryl-CoA reductase mRNA transcripts in goldfish (Carassius auratus)
Authors:Paolo Cocci  Gilberto MosconiFrancesco Alessandro Palermo
Affiliation:School of Biosciences and Veterinary Medicine, University of Camerino, Via Gentile III Da Varano, I-62032 Camerino, MC, Italy
Abstract:Epigallocatechin gallate (EGCG), the major active component of the green tea, has recently been found to inhibit 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCoAR) activity in vitro and to modulate lipogenesis in vivo. In this study we have evaluated the effects of short-term in vivo exposure to EGCG (6 μg g− 1 BW or 9 μg g− 1 BW) on hepatic HMGCoAR gene expression of goldfish (Carassius auratus). We initially characterized a partial sequence of goldfish HMGCoAR suggesting that the obtained fragment shares high similarity (> 92%) with other fish HMGCoAR sequences. Further, the HMGCoAR transcript was detected in all goldfish tissues (except muscle) but primarily in liver, brain and gonads; on the contrary, low expression levels were found in intestine, heart, gill, and kidney. Both EGCG doses significantly decreased hepatic HMGCoAR mRNA levels 180 min post-injection. HMGCoAR was also significantly down-regulated at 90 min after injection in fish treated with the highest dose of EGCG. Our results demonstrate that hepatic HMGCoAR gene expression is acutely responsive to short-term EGCG exposure in goldfish. This finding suggests a potential role of EGCG in transcriptional regulation of the rate-limiting enzyme in cholesterol synthesis.
Keywords:ABCA1, ATP-binding cassette transporter A1   BW, body weight   CYP7A1, cholesterol 7alpha-hydroxylase   EGCG, Epigallocatechin gallate   HMGCoAR, 3-hydroxy-3-methylglutaryl-CoA reductase   HMGCoAR-a, 3-hydroxy-3-methylglutaryl-CoA reductase isoform A   HMGCoAR-b, 3-hydroxy-3-methylglutaryl-CoA reductase isoform B   LXR, liver X receptor   MS-222, 3-aminobenzoic acid ethyl ester   NADPH, nicotinamide adenine dinucleotide phosphate   ORF, open reading frame   PPM, part per million   RT-PCR, real-time PCR   SEM, standard error of the mean   Srebf1, Sterol regulatory element-binding transcription factor 1   SREBP, Sterol regulatory element binding protein   SSD, sterol-sensing domain
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