24-脱氢胆固醇还原酶基因转基因小鼠血生化指标分析

The Effect of Dhcr24 on Blood Chemistry in the Dhcr24 Transgenic Mouse

目的建立全身表达24-脱氢胆固醇还原酶基因(Dhcr24)转基因小鼠动物模型,研究该基因过表达对小鼠代谢的影响。方法RT-PCR法克隆小鼠Dhcr24基因,把该基因插入CMV启动子下游,构建转基因表达载体,通过显微注射法建立Dhcr24转基因小鼠。PCR鉴定Dhcr24转基因小鼠的基因型,RT-PCR和Western Blot检测基因表达水平,血生化检测仪检测转基因小鼠血生化指标的改变。结果建立了2个不同表达水平的Dhcr24转基因小鼠品系,转入的Dhcr24基因在肝和脾组织中的表达高于内源的Dhcr24。血生化检测证实:乳酸脱氢酶(LDH)、总蛋白(TP)、白蛋白(Alb)和血肌酐(SCr)较野生型小鼠明显降低,而高密度脂蛋白胆固醇(HDL-c)和碱性磷酸酶(ALP)较野生型小鼠明显增加,并且Dhcr24转基因雌鼠的体重比野生型小鼠明显降低,均有显著差异。但Dhcr24转基因雄鼠各项指标与野生型小鼠相比没有显著差异。结论成功建立了全身表达Dhcr24转基因小鼠,并证实Dhcr24基因对雌性小鼠的体重和血生化指标,包括LDH,TP,Alb,SCr,HDL-c and ALP具有明显的影响。

Objective To establish the transgenic mouse of 24-dehydrocholesterol reductase gene（Dhcr24） to study the effect of Dhcr24 on metabolism in vivo.Method／ The mouse Dhcr24 was clone by RT-PCR and the sequence of the cDNA was confirmed by sequence analysis.The transgenic plasmid was constructed by inserting the Dhcr24 gene in the downstream of CMV promoter.The transgenic mice were generated by microinjection method and the genotype was detected by PCR.The expression levels of the gene were determined with RT-PCR and Western Blot.The blood biochemistry items were detected with a Photometer Reflotron Plus for blood following clinical procedure.Result／ Two transgenic C57BL/6J lines of Dhcr24 gene with different expression levels were established.The expression of the Dhcr24 gene was obviously detected in the liver and spleen.The transgenic mice showed obviously decreased levels of weight,lactic dehydrogenase（LDH）,total protein（TP）,Albumin（Alb） and serum creatinine（SCr） compared with that of the wild type mice,while the increased high-density lipoprotein cholesterol and increased alkaline phosphatase（ALP） were detected in the transgenic mice.Conclusion／ The DHCR24 transgenic mice were established successfully and indicated the Dhcr24 gene regulated body weight of the female mice and the blood chemistry included the levels of LDH,TP,Alb,SCr,HDL-c and ALP of female mice.