目的测定小鼠血清、肝脏及粪便中3H-胆固醇占经腹腔注射3H-胆固醇总量的百分比,探讨直接检测活体内巨噬细胞来源的胆固醇逆向转运方法。方法16只C57BL/6小鼠随机分为两组,分别经腹腔注射普通培养基(对照组)或乙酰化低密度脂蛋白及3H-胆固醇处理过的RAW264.7细胞悬液(实验组),单独笼养12 h和24 h后取血,酶法测定血清脂质;收集血清、肝组织、0~24 h粪便,用液闪计数仪测定各样本的3H-胆固醇含量(以占注射总量百分比计算)。结果24 h后实验组小鼠血清、肝脏和粪便中均可检测到3H的放射活性,分别占注射总量的2.810%±0.060%、0.680%±0.030%及0.690%±0.030%,且明显高于对照组。结论经腹腔注射乙酰化低密度脂蛋白及3H-胆固醇处理过的RAW264.7小鼠巨噬细胞悬液,24 h后可以分别检测到小鼠血清、肝组织、0~24 h粪便中3H-胆固醇含量,是一种直接检测小鼠体内巨噬细胞来源的胆固醇逆向转运的可行方法。
Aim To set up a method to measure the rate of reverse cholesterol transport from macrophage to feces in vivo,serum lipid profiles of mice was tested and the 3 H-contents was quantitated in serum,liver and feces in mice after 24 hours intraperitoneally were injected macrophages which were labeled with 3 H cholesterol.Methods 16 C57BL/6 mice were divided into two groups at random and one group was injected intraperitoneally with RAW264.7 macrophages which were loaded with cholesterol by incubation with acetylated low density lipoprotein(acLDL),labeled with 3 H-cholesterol.As a control,another group was injected with culture media instead.Serum lipid profiles were quantitated by enzymatic method;serum and liver tissues were harvested at 24 hours,feces were collected(0~24 h),and all were analyzed for tracer counts(all were expressed as the percent of the total injected counts per minute).Results After being injected intraperitoneally 24 h with DMEM or RAW264.7 macrophages,the serum lipid profiles of two groups have no difference(P〉 0.05).3 H-cholesterol could be detected in the plasma,liver,and feces of the mice in tested group,and the percents of the total injected counts per minute were 2.810%± 0.060%,0.680% ±0.030% and 0.690%±0.030% respectively,also were higher than that of control group.Conclusions After being injected intraperitoneally with 3 H-cholesterol-labled RAW264.7 macrophages,the radioactivity could be detected in the plasma,liver,and feces of the mice.This method could be taken as a kind of feasible way to measure the rate of reverse cholesterol transport from macrophages into feces.