【摘要】：為建立從江香豬(Sus scrofa)脂肪前體細胞的體外培養(yǎng)模式,分析從江香豬肌內(nèi)和皮下脂肪前體細胞分化過程中相關(guān)基因的表達差異。采集5日齡從江香豬背最長肌和皮下脂肪組織,利用膠原酶消化法,分別從此兩種組織中分離獲得脂肪前體細胞。經(jīng)誘導分化后,采用q RT-PCR技術(shù)檢測細胞分化過程中脂肪合成與分解基因:轉(zhuǎn)錄因子過氧化物酶體增殖物激活受體γ(peroxisome proliferator activated receptorγ,PPARγ)、脂蛋白脂酶(lipoprotein lipase,LPL)、脂肪酸合成酶(fatty acid synthase,FAS)、乙酰輔酶A羧化酶(acetyl-Co A carboxylase,ACC)、甘油三酯水解酶(adipose triacylglycerol lipase,ATGL)、激素敏感脂酶(hormone-sensitive lipase,HSL)的時序表達情況。成功構(gòu)建從江香豬脂肪前體細胞的體外培養(yǎng)模式。通過q RT-PCR分析顯示,在肌內(nèi)脂肪前體細胞中,PPARγ、FAS、ACC m RNA表達水平在整個分化早期72 h時均呈現(xiàn)較高表達,顯著高于0 h的表達水平(P0.05);LPL m RNA表達水平在分化早期各階段無明顯差異(P0.05),但在144 h時出現(xiàn)顯著的大幅度上調(diào),極顯著高于其他各階段表達水平(P0.01);ATGL、HSL m RNA表達水平在整個分化階段均呈現(xiàn)"升高-降低-升高"的表達規(guī)律。在皮下脂肪前體細胞中,PPARγ基因在誘導分化72 h的表達水平最高,極顯著高于其他幾個階段的表達水平(P0.01);LPL、FAS、ACC、ATGL、HSL m RNA表達水平在整個分化早期48和72 h均呈現(xiàn)較高表達,極顯著高于0 h的表達水平(P0.01),之后均下降。在離體培養(yǎng)條件下,各脂肪細胞分化相關(guān)基因在肌內(nèi)脂肪前體細胞的分化后期,表現(xiàn)較活躍;而各脂肪細胞分化相關(guān)基因在皮下脂肪前體細胞的分化早期,表現(xiàn)較活躍。研究結(jié)果表明從江香豬皮下脂肪的形成可能遠遠早于肌內(nèi)脂肪,對進一步研究從江香豬脂肪沉積機制具有重要意義。
[Abstract]:In order to establish an in vitro culture model of (Sus scrofa) adipocyte precursor cells in Congjiang Xiang pig, the difference of expression of related genes in the differentiation of intramuscular and subcutaneous fat precursor cells of Congjiang Xiang pig was analyzed. The longissimus dorsi muscle and subcutaneous adipose tissue were collected and digested with collagenase to isolate adipose precursor cells from the two tissues. After induced differentiation, Using Q RT-PCR technique to detect adipose synthesis and decomposition genes during cell differentiation: transcription factor peroxisome proliferator activated receptor 緯 (peroxisome proliferator activated receptor 緯 PPAR 緯, lipoprotein lipase (lipoprotein lipase,LPL), fatty acid synthase (fatty acid synthase,FAS, acetyl coenzyme A carboxylase The temporal expression of acetyl-Co A carboxylase,ACC, triglyceride hydrolase (adipose triacylglycerol lipase,ATGL and hormone sensitive lipase (hormone-sensitive lipase,HSL). The in vitro culture model of fat precursor cells of Congjiang Xiang pig was successfully constructed. Q RT-PCR analysis showed that the expression of PPAR- 緯 FASACC m RNA in intramuscular adipocytes was higher at 72 h after differentiation. The expression level of LPLM RNA was significantly higher than that of 0 h (P0.05). There was no significant difference in the expression of LPLM RNA in the early stage of differentiation (P0.05), but it was significantly up-regulated at 144h. The expression level of HSLm RNA was significantly higher than that of other stages (P0.01). The expression level of PPAR 緯 gene in subcutaneous adipose progenitor cells was the highest at 72 h after differentiation, which was significantly higher than that in other stages (P0.01). The expression level was significantly higher than 0 h (P0.01), and then decreased. In vitro, the adipocyte differentiation related genes were active in the later stage of the differentiation of intramuscular adipocytes, while the adipocyte differentiation related genes were active in the early stage of subcutaneous adipose progenitor cell differentiation. The results showed that the formation of subcutaneous fat in Congjiang Xiang pig may be much earlier than that of intramuscular fat, which is of great significance to further study the mechanism of fat deposition in Congjiang Xiang pig.
【作者單位】： 貴州大學高原山地動物遺傳育種與繁殖省部共建教育部重點實驗室;貴州大學動物科學學院;貴州省動物遺傳育種與繁殖重點實驗室;貴州大學生命科學學院;
【基金】：國家科技支撐計劃(No.2015BAD03B02-3) 貴州畜禽種質(zhì)資源保存創(chuàng)新與利用(黔科合重大專項字(2013)6008) 貴州大學研究生創(chuàng)新基金(研農(nóng)2016021)
【分類號】：S828

【相似文獻】

相關(guān)期刊論文 前1條

1 馮祖蔭;;黃脂豬的鑒別與處理方法[J];畜牧與獸醫(yī);1983年06期

相關(guān)博士學位論文 前2條

1 褚敏;營養(yǎng)脅迫條件下牦牛皮下脂肪和背肌差異microRNAs的篩選與鑒定[D];中國農(nóng)業(yè)科學院;2015年

2 褚維偉;豬肌內(nèi)脂肪和皮下脂肪的糖皮質(zhì)激素敏感性差異及其機理初探[D];南京農(nóng)業(yè)大學;2016年

，

本文編號：2223764