本文選題：hNPCL + 過(guò)表達(dá)��； 參考：《浙江農(nóng)業(yè)學(xué)報(bào)》2017年10期

【摘要】：利用體細(xì)胞核移植技術(shù)將構(gòu)建的肝過(guò)表達(dá)h NPC1L1的小型豬胎兒成纖維細(xì)胞作為核供體細(xì)胞,構(gòu)建了肝過(guò)表達(dá)h NPC1L1轉(zhuǎn)基因巴馬小型豬,PCR和RT-PCR鑒定表明h NPC1L1特異性地表達(dá)于肝組織。對(duì)轉(zhuǎn)基因巴馬小型豬進(jìn)行了初步研究,脂質(zhì)過(guò)氧化標(biāo)記物丙二醛(MDA)檢測(cè)表明:轉(zhuǎn)基因豬MDA含量(7.51±0.09)nmol·m L-1顯著高于非轉(zhuǎn)基因豬(3.56±0.16)nmol·m L-1(P0.001),說(shuō)明肝組織發(fā)生了嚴(yán)重的脂質(zhì)過(guò)氧化。超氧化物歧化酶(SOD)活性檢測(cè)表明:SOD活性為(131±4.61)U·mg-1顯著低于非轉(zhuǎn)基因豬[(229.2±4.39)U·mg-1](P0.001),說(shuō)明肝臟清除自由基和抗氧化能力大大下降。試驗(yàn)揭示出在肝臟過(guò)表達(dá)h NPC1L1導(dǎo)致嚴(yán)重的脂質(zhì)過(guò)氧化,表明NPC1L1在脂代謝紊亂和肝臟疾病中扮演著重要的角色,有望成為肝臟疾病治療新的靶標(biāo)。
[Abstract]:Using somatic cell nuclear transfer technique, the constructed porcine fetal fibroblasts which over-expressed hNPC1L1 were used as nuclear donor cells. PCR and RT-PCR analysis showed that hNPC1L1 was specifically expressed in the liver tissue of HNPC1L1 transgenic Bama mini-pig. The malondialdehyde (MDA), a lipid peroxidation marker, was detected in transgenic Bama miniature pigs. The results showed that the content of MDA in transgenic pigs (7.51 鹵0.09) nmol m L-1) was significantly higher than that in non-transgenic pigs (3.56 鹵0.16) nmol m L-1 (P0.001), indicating that serious lipid peroxidation occurred in liver tissue. The activity of superoxide dismutase (SOD) was (131 鹵4.61) U mg-1 significantly lower than that of non-transgenic pigs [(229.2 鹵4.39) U mg-1] (P0.001). The results showed that overexpression of hNPC1L1 in the liver resulted in severe lipid peroxidation, suggesting that NPC1L1 plays an important role in lipid metabolism disorders and liver diseases, and may become a new target for the treatment of liver diseases.
【作者單位】： 吉林化工學(xué)院生物與食品工程學(xué)院;韶關(guān)學(xué)院英東生命科學(xué)院粵北生豬生產(chǎn)及疫病防控協(xié)同創(chuàng)新發(fā)展中心;吉林農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)技術(shù)學(xué)院;吉林大學(xué)動(dòng)物醫(yī)學(xué)學(xué)院;
【基金】：國(guó)家科技支撐計(jì)劃(2011BA115B02)
【分類(lèi)號(hào)】：R575

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