肝过表达hNPC1L1促进脂质过氧化

Overexpression of human Niemann-Pick C1-Like 1 in livers accelerates lipid peroxidation

利用体细胞核移植技术将构建的肝过表达h NPC1L1的小型猪胎儿成纤维细胞作为核供体细胞,构建了肝过表达h NPC1L1转基因巴马小型猪,PCR和RT-PCR鉴定表明h NPC1L1特异性地表达于肝组织。对转基因巴马小型猪进行了初步研究,脂质过氧化标记物丙二醛(MDA)检测表明:转基因猪MDA含量(7.51±0.09)nmol·m L-1显著高于非转基因猪(3.56±0.16)nmol·m L-1(P<0.001),说明肝组织发生了严重的脂质过氧化。超氧化物歧化酶(SOD)活性检测表明:SOD活性为(131±4.61)U·mg-1显著低于非转基因猪[(229.2±4.39)U·mg-1](P<0.001),说明肝脏清除自由基和抗氧化能力大大下降。试验揭示出在肝脏过表达h NPC1L1导致严重的脂质过氧化,表明NPC1L1在脂代谢紊乱和肝脏疾病中扮演着重要的角色,有望成为肝脏疾病治疗新的靶标。

The transgenic Bama miniature pigs in which human Niemann-Pick C1-Like 1（ h NPC1 L1） was overexpressed in the livers were constructed by using somatic cell nuclear transfer technique with the minipig fetal fibroblast cells as the nuclear donor cells. The h NPC1 L1 was specifically expressed in transgenic liver tissues as revealed by PCR and reverse transcriptase（ RT）-PCR analysis. A preliminary study on transgenic Bama miniature pigs was carried out,and the marker malondialdehyde（ MDA） of lipid peroxidation was detected. The result showed that the level of MDA in transgenic pigs [（ 7. 51 ± 0. 09） nmol·m L-1]was significantly higher than that in non-transgenic pigs[（ 3. 56 ± 0. 16） nmol·m L-1]（ P 0. 001）,but SOD level in transgenic pigs [（ 131 ± 4. 61） U·mg-1]was significantly lower than that in non-transgenic pigs [（ 229. 2 ± 4. 39） U·mg-1]（ P 0. 001）. All of these suggested that severe lipid peroxidation occurred in the liver and the oxygen free radical scavenging and antioxidant capacity weresignificantly reduced. This study revealed that overexpression of NPC1 L1 in the liver led to severe lipid peroxidation,which indicated that NPC1 L1 played an important role in lipid metabolism disorders and liver diseases and could be a new target for liver disease treatment.