Analysis of the induction mechanisms of atherosclerosis using C-reactive protein transgenic rabbits

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In this study, we made plivLE6-hCRP construct which inserted human C-reactive protein (h-CRP) cDNA in plivHEG/LE6 vector to express high h-CRP in liver specifically, and it was injected into rabbit fertilized egg pronuclei by microinjection method. After microinjection, 453 embryos were transplanted in fallopian tubes of the foster rabbits induced pseudopregnancy by hCG injection and they gave birth to 47 pups in all. PCR analysis revealed transgene was introduced in 5 of 47 pups, and we confirmed h-CRP expression in plasma of 2 rabbits (Line C205 and C215) survived until weaning. After expand of the each colony of C205 and C215, we examined the expression level of h-CRP in each transgenic line, and we confirmed that C205 and C215 showed 7.3±1.0 and 122.0±13.4 mg/dL, respectively. In comparison of plasma lipid profile such as total cholesterol, HDL-cholesterol and triglyceride, there were no differences between transgenic and non-transgenic rabbits.For analysis of the induction mechanisms of atherosclerosis, transgenic and non-transgenic rabbits were fed cholesterol-rich diet for 16 weeks. The concentration of plasma cholesterol was measured weekly and adjusted to same level between 2 groups by changing the amount of cholesterol in the diet (0.075-0.3%). Cholesterol loading was finished already, and we have collected aortas and hearts from all rabbits for pathological analysis. At present, more detailed histopathological analysis is doing for preparation of manuscript.