肝过表达hNPC1L1转基因巴马小型猪的初步研究

Preliminary Study on Transgenic Bama Miniature Pig with Liver Over-expression of hNPC1L1

研究利用体细胞核移植技术构建了肝过表达h NPC1L1转基因巴马小型猪,由PCR和RT-PCR鉴定表明h NPC1L1在肝组织中特异性地表达。我们对转基因巴马小型猪进行了初步研究,对血清和肝匀浆中的TC、TG、LDL-C、HDL-C、AST、ALT和FFA进行了检测,结果表明转基因猪血清FFA(637.7±9.53)μmol/L显著高于非转基因猪血清FFA(540.4±25.46)μmol/L(p<0.01),转基因猪血清AST(200.8±22.31)U/L显著高于非转基因猪血清AST(92.4±15.55)U/L(p<0.01)。另外,我们对肝中与脂代谢密切相关的基因表达情况进行Real-time PCR检测的结果表明,转基因猪SREBP-1c(3.31±0.18,p<0.05)、FAS(4.49±4.21,p<0.01)显著高于非转基因猪。本研究发现NPC1L1在与脂代谢紊乱密切相关的肝脏疾病中发挥着主导作用,它能通过对脂合成相关基因转录的调控,引发脂代谢紊乱以及肝脏疾病。这说明对NPC1L1的研究有望成为治疗肝脏疾病的新靶标。

In this research, Transgenic Bama miniature pigs which human Niemann-Pick C1-Like 1 was overexpressed in the livers were constructed by using somatic cell nuclear transfer technique. The hNPCIL1 was specifically expressed in liver tissues by the identification of PCR and RT-PCR analysis. A preliminary study on transgenic Bama miniature pigs was carried out and the levels of TC, TG, LDL-C, HDL-C, AST, ALT, FFA in serum and liver homogenate were measured. The result showed that the level of FFA in the serum transgenic pigs （（637.7±9.53） μmol/L） was significantly higher than that in non-transgenic pigs （（540.4±25.46） μmol/L） （P〈0.01）. The level of AST （（200.8±22.31） U/L） in the serum transgenic pigs was significantly higher than that in non- transgenic pigs （（92.4±15.55） U/L） （p〈0.01）. Besides, Real-time PCR was used to detect the expression of genes closely related to cholesterol and fat metabolism. The results showed that the SREBP- 1c （3.31±0.18, p 〈0.05） and FAS （4.49 ±4.21, p〈0.01） of transgenic pigs was significantly higher than that in non-transgenic pigs. Our findings revealed that NPC1L1 plays an important role in liver diseases which closely related to lipid metabolism disorders. NPC1L1 can regulate the transcription of certain genes involved in lipid synthesis and leads to lipid metabolism disorders and liver disease. This indicates that the study of NPC1L1 is expected to be a new target for liver disease treatment.