抑制TGF-β/Smads通路对Hyp促进浅色黄姑鱼鱼鳔胶原蛋白沉积的影响  

Effects of inhibiting TGF-β/Smads pathway on hydroxyproline promotion of collagen deposition in swim bladder of Chu’s croaker (Nibea coibor)

作　　者：荣华[1,2] 杨石坤 杜有家 豆腾飞 黄英[1] 武祥伟[1,2] 史庆超 王晓雯[1,2] RONG Hua;YANG Shikun;DU Youjia;DOU Tengfei;HUANG Ying;WU Xiangwei;SHI Qingchao;WANG Xiaowen(College of Animal Science and Technology,Yunnan Agricultural University,Kunming,Yunnan 650201,China;Key Laboratory of Protection and Sustainable Utilization of Plateau Fishery Resources in University of Yunnan Province,Yunnan Agricultural University,Kunming,Yunnan 650201,China;Key Laboratory of Sichuan Province for Fishes Conservation and Utilization in the Upper Reaches of the Yangtze River,Neijiang Normal University,Neijiang,Sichuan 641112,China)

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201 [2]云南农业大学云南省高校高原渔业资源保护与可持续利用重点试验室,云南昆明650201 [3]内江师范学院长江上游鱼类资源保护与利用四川省重点试验室,四川内江641112

出　　处：《西北农林科技大学学报（自然科学版）》2022年第9期1-10,共10页Journal of Northwest A&F University(Natural Science Edition)

基　　金：云南省农业基础研究联合专项面上项目(2018FG001-044);长江上游鱼类资源保护与利用四川省重点实验室开发基金项目(NJTCCJSYSYS12);四川省科技厅重点研发项目(2019ZYZF0099)。

摘　　要：【目的】探讨Hyp对浅色黄姑鱼鱼鳔胶原蛋白沉积的影响及其作用机制,为优质鱼胶的生产提供营养调控策略。【方法】将300尾浅色黄姑鱼(体质量(11.621±0.154) g/尾)随机分为Hyp空白组、Hyp添加组、SIS3干扰组、氧化苦参碱干扰组,每组3个重复,每重复25尾鱼,Hyp空白组饲喂日粮中不添加Hyp,其他3组均饲喂添加9.70 g/kg(前期试验确定的最佳添加量)L-羟脯氨酸的日粮,进行为期8周的饲养试验,SIS3、氧化苦参碱干扰组试验鱼在1月后分别腹腔注射转化生长因子(TGF-β)/Smads通路抑制剂SIS3和氧化苦参碱注射液100μL/尾(SIS3和氧化苦参碱的计量分别为2和10 mg/kg)进行干扰试验,每周注射1次,直至试验结束。试验结束后,测定各处理组中浅色黄姑鱼生长指标(特定生长率、饲料系数、鳔体比、存活率、肥满度)、鱼鳔胶原蛋白含量,以及胶原蛋白代谢相关基因(COL1A1、COL1A2、P4Hα(Ⅰ)、P4Hα(Ⅱ)、P4Hα(Ⅲ))和TGF-β/Smads通路关键基因(TGF-β、TGF-βRT、Smad2、Smad3、Smad4、Smad7)的表达,分析鱼鳔中胶原蛋白差异化沉积是否受TGF-β/Smads通路的调控。【结果】Hyp能显著促进浅色黄姑鱼生长及鱼鳔中胶原蛋白沉积,能显著上调COL1A1、COL1A1基因的表达,显著下调P4Hα(Ⅲ)的表达;SIS3显著降低了鱼鳔胶原蛋白沉积;SIS3和氧化苦参碱对胶原蛋白代谢相关基因及TGF-β/Smads信号通路关键基因表达无显著影响。【结论】TGF-β/Smads信号通路可能不是调控Hyp促进鱼鳔胶原蛋白沉积的主要途径。Hyp作为胶原蛋白分解产物,其促进胶原蛋白沉积可能与其抑制胶原蛋白降解有关。【Objective】 This study investigate the effects of dietary Hyp supplementation on swim bladder collagen synthesis and its possible molecular mechanism to provide nutrition regulation strategies for the production of high-quality fish gelatin.【Method】 Three hundred Chu’s croaker(Nibea coibor) at body weight of(11.621±0.154) g were randomly divided into 4 groups of Hyp blank group,Hyp addition group,SIS3 interference group and oxymatrine interference group,with 3 replicates per group and 25 fish per replicate.The fish in the Hyp blank group was fed the diet with no Hyp added,and the others were fed the diet supplemented with 9.70 g/kg L-hydroxyproline(the optimal addition amount determined by previous experiment) for eight weeks.After feeding N.coibor for one month,SIS3 interference group and oxymatrine interference group were injected by intraperitoneal with specific inhibitor of transforming growth factor beta(TGF-β)/Smads pathway SIS3 and oxymatrine for 100 μL/tail(doses of SIS3 and oxymatrine were 2 and 10 mg/kg,respectively) once a week until the end of the experiment.At the end of experiment,the collagen content in swim bladder of N.coibor and the growth performance indexes such as specific growth rate(SGR),feed coefficient(FC),swim bladder somatic index(SBSI),survival rate(SR) and condition factor(CF) were determined.The expressions of genes related to collagen metabolism(COL1A1,COL1A2,P4Hα(Ⅰ),P4Hα(Ⅱ) and P4Hα(Ⅲ)) and key genes of TGF-β/Smads pathway(TGF-β,TGF-βRT,Smad2,Smad3,Smad4 and Smad7) were measured.Then,it was analyzed that whether the differential deposition of collagen in the swim bladder was regulated through the TGF-β/Smads pathway.【Result】 Hyp significantly increased the growth and collagen deposition of N.coibor and up-regulated the expression of type Ⅰ collagen α1/2 genes(COL1A1 and COL1A2).The expression of P4Hα(Ⅲ) gene was down-regulated significantly.SIS3 significantly reduced collagen deposition in swim bladder.However,SIS3 and oxymatrine had no significa

关 键 词：羟脯氨酸 TGF-β/Smads通路 浅色黄姑鱼 鱼鳔 胶原蛋白 

分 类 号：S965.325[农业科学—水产养殖]