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| 论文摘要： | Collagen serves as one of determinants of fish muscle quality. Timp2 is a key enzyme in collagen metabolism. Its role in grass carp (Ctenopharyngodon idellus) and related nutritional regulation mechanism are unknown. Given the important role of copper as a micronutrient in collagen biosynthesis, it was hypothesized that Cu2+ up-regulates timp2 expression through the TGF beta/Smads signaling pathway, thus modulating grass carp collagen content. In the present study, this hypothesis validation was performed through parallel experiments using the TGF-beta/Smads inhibitor SB431542 and timp2-specific siRNA (sitimp2) in cultured fibroblasts. Results showed that both sitimp2 and SB431542 treatment decreased the collagen content (to 90.2 % +/- 0.52 % and 61.8 % +/- 1.59 % of control, respectively), down-regulated timp2 by 53.8 % +/- 1.30 % and 11.3 % +/- 1.91 % relative to control) and up-regulated mmp2 by 33.1 % +/- 9.70 % and 10.9 % +/- 2.51 % relative to control). Additionally, SB431542 inhibited the TGF beta/Smads signaling pathway (P < 0.05). Coversely, Cu2+ supplementation significantly increased the collagen content, up-regulated timp2 expression and enhanced TGF beta/Smads signaling pathway activity, while simultaneously suppressing mmp2 P < 0.05). Futhermore, after sitimp2 or SB431542 treatment, 10 mu M Cu2+ reversed the changes in the relevant indicators induced by both sitimp2 and SB431542 treatment (P < 0.05). These results confirmed the hypothesis that Cu2+ enhanced collagen deposition through TGF beta/Smads-mediated up-regulation of timp2. The present study provided novel mechanistic insights into copper-dependent collagen regulation and suggested that dietary copper supplementation could be a practical nutritional strategy to improve muscle texture and product quality in farmed grass carp. |
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