脂肪酸脱氢酶基因在兔体内表达的生物学效应研究

目的研究脂肪酸脱氢酶基因在兔子体内的生物效应,探索不饱和脂肪酸与兔子免疫的相互作用。方法用壳聚糖(CS)和壳聚糖的聚乙烯亚胺(PEI)、聚乙二醇(PEG)修饰产物(mPEG-O-CS-PEI),对含有脂肪酸脱氢酶基因的真核表达质粒VRMFat-1进行包裹,肌肉注射新西兰白兔,每两周采取抗凝血,检测血液中免疫细胞数量的变化情况,利用气相色谱仪检测动物组织中脂肪酸的含量变化,并通过荧光定量PCR检测免疫细胞内免疫相关基因的表达情况。结果两组实验兔组织中的n-3不饱和脂肪酸含量显著高于对照组(P<0.05),其TLR4、CD3、IL-4和IL-6基因表达水平显著升高(P<0.05)。结论转入的脂肪酸去饱和脱氢酶基因能够在实验兔体内正常表达,初步证明不饱和脂肪酸对兔子的固有免疫和获得性免疫机能均有明显的增强作用。

Studies of Biological Effects of the Expression of Fatty Acid Desaturase Gene on Rabbits

Objective To study the biological effects of the expression of fatty acid desaturase gene in vivo on the rabbit,and explore the relationship between unsaturated fatty acids and immune system of rabbit.Methods The eukaryotic expression plasmid VRMFat-1 were packed with chitosan(CS),chitosan modified with polyethylenimine(PEI) and polyethylene glycol(PEG),then New Zealand white rabbits were intramuscularly injected with the packed plasmids.The blood of rabbits were collected every two weeks to test the changes of immune cells,fatty acid content in animal tissues and the expression level of immune genes by gas chromatography and quantitative RT-PCR.Results It was found that the n-3 polyunsaturated fatty acids in two experimental groups was significantly higher than the control group(P<0.05).Meanwhile,the expression of TLR4,CD3,IL-4 and IL-6 gene were significantly enhanced/upregulated in treated experimental group(P<0.05).Conclusion The transferred fatty acid desaturase gene can be effectively expressed in experimental rabbits,and improve the content of unsaturated fatty acids that can significantly enhance the innate and acquired immune function.