黄连解毒汤对apoE基因敲除小鼠单核细胞发育的影响

Effect of Huanglian Jiedu Decoction on Monocyte Development in apoE Gene Knockout Mice

目的观察野生型C57BL/6小鼠和apoE基因敲除(apoE^(-/-))小鼠外周血单核细胞(monocyte,Mo)发育情况及黄连解毒汤对apoE^(-/-)小鼠外周血Mo发育的影响。方法 4、8、12、16周龄雌性C57BL/6为不同周龄对照组,4、8、12、16周龄雌性apoE^(-/-)小鼠为不同周龄高脂血症组。4周龄C57BL/6雌性小鼠为空白组,4周龄apoE^(-/-)雌性小鼠采用配对比较法随机分为对照组、西药组和中药组,每组5只。给药小鼠根据体重给予临床等效剂量,西药组灌胃阿托伐他汀[10mg/(kg·d)],中药组灌胃黄连解毒汤[5g/(kg·d)]。每周检测体重,4周后检测TG、TC、LDL-C和HDL-C水平、Mo及其亚型Ly6chi比例变化。结果与同种小鼠4周龄组比较,C57BL/6 16周龄组Mo比例降低(P<0.05),apoE^(-/-)8周龄小鼠TC、TG、Ly6c^(hi)比例升高(P<0.05),Mo比例降低(P<0.05);12周龄小鼠TC、TG和LDL-C升高(P<0.05),16周龄小鼠TC、TG、LDL-C及HDL-C升高(P<0.05,P<0.01)。与同种小鼠8周龄组比较,C57BL/6 16周龄组Mo比例降低(P<0.05);12周龄apoE^(-/-)小鼠TC和LDL-C升高(P<0.05),16周龄小鼠TC和HDL-C升高(P<0.05,P<0.01)。与同周龄C57BL/6组比较,4、8周龄apoE^(-/-)小鼠TC、TG升高(P<0.01),HDL-C降低(P<0.01),12、16周龄小鼠TC、TG、LDL-C升高,HDL-C降低(P<0.05,P<0.01);apoE^(-/-)4周龄小鼠Mo比例升高(P<0.05),8周龄小鼠Mo、Ly6c^(hi)比例升高(P<0.05)。与空白组比较,对照组小鼠干预后TC、TG、LDL-C、Mo及Ly6chi比例升高(P<0.01,P<0.05),HDL-C降低(P<0.01)。与对照组比较,西药组和中药组小鼠体重增长降低(P<0.05),中药组Ly6chi比例降低(P<0.05)。结论在发育过程中apoE^(-/-)小鼠的血脂水平不仅与周龄相关,由其引发的天然免疫系统的发育改变也与周龄有关。发育早期用黄连解毒汤干预apoE^(-/-)小鼠可纠正这种独特免疫效应。

Objective To observe monocyte（Mo）development in wild type C57BL/6 mice and apoE gene knockout（apoE^-/-）mice,and to evaluate the immuno-regulatory effect of Huanglian Jiedu Decoction（HJD）on peripheral Mo development in apoE^-/- mice.Methods Four,8,12,and16 weeks old female C57BL/6 mice were set up as control groups of different ages,while 4,8,12,and 16 weeks old female apoE^-/- mice were set up as hyperlipidemia groups of different ages.Four-week old female C57BL/6 mice were recruited as a blank group.Four-week old female apoE^-/- mice were randomly divided into the control group,the Western medicine group,and the Chinese medicine group by paired comparison,5in each group.Equivalent clinical dose was administered to mice according to bodyweight.Mice in the Western medicine group were administered with Atrovastatin at the daily dose of 10 mg/kg by gastrogavage,while those in the Chinese medicine group were administered with HJD at the daily dose of 5 g/kg by gastrogavage.Body weight was detected each week.After 4 weeks blood lipids levels（such as TG,TC,LDL-C,and HDLC）,and the proportions of Mo and Ly6c^hi were detected.Results Compared with4-week-old homogenic mice,the proportion of Mo decreased in16-week-old C57BL/6 mice（P〈0.05）.Levels of TC and TG,and the proportion of Ly6 c^hi subtype increased,but the proportion of Mo decreased in8-week-old apoE^-/- mice（P〈0.05）.Levels of TC,TG,and LDL-C increased in 12-week-old apoE^-/- mice（P〈0.05）.Levels of TC,TG,LDL-C,and HDL-C increased in 16-week-old apoE^-/- mice（P〈0.05,P〈0.01）.Compared with 8-week-old homogenic mice,the proportion of Mo decreased in 16-week-old C57BL/6 mice（P〈0.05）;levels of TC and LDL-C increased in12-week-old apoE^（-/-）mice（P〈0.05）;levels of TC and HDL-C increased in 16-week-old apoE^-/- mice（P〈0.05,P〈0.01）.Compared with C57BL/6 mice of the same age,TC and TG increased,HDL-C decreased（P〈0.01）in4-and8-week-old apoE^-/- mice（P〈0.01）;levels of TC,TG,LDL-C increased,and HDL-C level decreased in 12-and 16-week-old apoE^-/- mice（P〈0.05,P〈0.01）;the proportion of Mo increased in 4-week-old apoE^-/-mice（P〈0.05）;proportions of Mo and Ly6c^hi increased in 8-week-old apoE^-/- mice（P〈0.05）.Compared with the blank control group,levels of TC,TG,and LDL-C,proportions of Mo and Ly6c^hi increased（P〈0.01,P〈0.05）,but HDL-C level decreased（P〈0.01）in the control group after intervention.Compared with the control group,body weight gained less in the Western medicine group and the Chinese medicine group（P〈0.05）;the proportion of Ly6 chi subtype decreased in the Chinese medicine group（P〈0.05）.Conclusions In development process blood lipids levels in apoE^-/- mice are not only associated with age.Blood lipids levels induced growth changes in natural immune system are also correlated with age.In early stage of lipids development HJD intervention could correct this special immune disorder in apoE^-/- mice.