發(fā)布時(shí)間：2018-09-05 08:41

【摘要】：為建立從江香豬(Sus scrofa)脂肪前體細(xì)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(xì)胞分化過(guò)程中相關(guān)基因的表達(dá)差異。采集5日齡從江香豬背最長(zhǎng)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(xì)胞。經(jīng)誘導(dǎo)分化后,采用q RT-PCR技術(shù)檢測(cè)細(xì)胞分化過(guò)程中脂肪合成與分解基因:轉(zhuǎn)錄因子過(guò)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(shí)序表達(dá)情況。成功構(gòu)建從江香豬脂肪前體細(xì)胞的體外培養(yǎng)模式。通過(guò)q RT-PCR分析顯示,在肌內(nèi)脂肪前體細(xì)胞中,PPARγ、FAS、ACC m RNA表達(dá)水平在整個(gè)分化早期72 h時(shí)均呈現(xiàn)較高表達(dá),顯著高于0 h的表達(dá)水平(P0.05);LPL m RNA表達(dá)水平在分化早期各階段無(wú)明顯差異(P0.05),但在144 h時(shí)出現(xiàn)顯著的大幅度上調(diào),極顯著高于其他各階段表達(dá)水平(P0.01);ATGL、HSL m RNA表達(dá)水平在整個(gè)分化階段均呈現(xiàn)"升高-降低-升高"的表達(dá)規(guī)律。在皮下脂肪前體細(xì)胞中,PPARγ基因在誘導(dǎo)分化72 h的表達(dá)水平最高,極顯著高于其他幾個(gè)階段的表達(dá)水平(P0.01);LPL、FAS、ACC、ATGL、HSL m RNA表達(dá)水平在整個(gè)分化早期48和72 h均呈現(xiàn)較高表達(dá),極顯著高于0 h的表達(dá)水平(P0.01),之后均下降。在離體培養(yǎng)條件下,各脂肪細(xì)胞分化相關(guān)基因在肌內(nèi)脂肪前體細(xì)胞的分化后期,表現(xiàn)較活躍;而各脂肪細(xì)胞分化相關(guān)基因在皮下脂肪前體細(xì)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(yuǎn)遠(yuǎn)早于肌內(nèi)脂肪,對(duì)進(jìn)一步研究從江香豬脂肪沉積機(jī)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(xué)高原山地動(dòng)物遺傳育種與繁殖省部共建教育部重點(diǎn)實(shí)驗(yàn)室;貴州大學(xué)動(dòng)物科學(xué)學(xué)院;貴州省動(dòng)物遺傳育種與繁殖重點(diǎn)實(shí)驗(yàn)室;貴州大學(xué)生命科學(xué)學(xué)院;
【基金】：國(guó)家科技支撐計(jì)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(zhuān)項(xiàng)字(2013)6008) 貴州大學(xué)研究生創(chuàng)新基金(研農(nóng)2016021)
【分類(lèi)號(hào)】：S828

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