雷公藤内酯醇对ApoE-/-小鼠动脉粥样硬化的作用研究

Effect of triptolide on atherosclerosis of ApoE knock-out mice

目的研究不同剂量雷公藤内酯醇对ApoE-/-小鼠动脉粥样硬化(Atherosclerosis,AS)的治疗效果。方法将4只10周龄雄性ApoE-/-小鼠作为模型组,4只10周龄雄性C57BL/6J小鼠作为对照组,适应性喂养1周后处死,取主动脉标本做HE染色。另将10周龄雄性ApoE-/-小鼠32只作为实验组,按每天每千克体重给药量随机分为4组:50μg/(kg·d)、75μg/(kg·d)组、100μg/(kg·d)、空白对照组,每组8只。10周龄雄性C57BL/6J小鼠8只作为阴性对照组。给药3周后处死小鼠,取主动脉标本做HE染色;收集血清,测定IL-10、IL-12含量。结果 (1)在11周龄时两组主动脉HE染色Roberts&Thompson方法评分分别为:模型组[(5.250±0.500)分]>对照组[(0.500±0.577)分],差异有统计学意义。(2)经不同浓度雷公藤内酯醇处理3周后,各组主动脉HE染色Roberts&Thompson方法评分分别为:空白对照组[(6.500±0.189)分]>100μg/(kg·d)组[(4.625±0.183)分]>50μg/(kg·d)组[(3.375±0.183)分]>75μg/(kg·d)组[(1.375±0.183)分]>阴性对照组[(0.000±0.000)分],差异有统计学意义。(3)75μg/(kg·d)组血清中IL-10浓度升高最明显,与各组比较差异均有统计学意义(P<0.001);各组血清中IL-12浓度均较空白对照组降低,其中75μg/(kg·d)组降低最明显,两者之间的差异有统计学意义(P<0.01)。结论 (1)在11周龄时,ApoE-/-小鼠能形成AS模型。(2)不同浓度雷公藤内酯醇处理3周后,ApoE-/-小鼠AS病变有不同程度缓解,其中75μg/(kg·d)浓度下效果最佳。(3)雷公藤内酯醇能够上调IL-10、下调IL-12的表达水平,抑制炎症反应,减轻粥样斑块的形成,此可能为免疫抑制剂抗动脉粥样硬化的机制之一。

Objective To explore the effect of different dose of triptolide on atherosclerosis of ApoE knock-out mice. Method Four male ApoE-/- mice （10 weeks old） and four male C57BL/6J mice （10 weeks old） were selected as model group and control group respectively. They were sacrificed after feeding with normal food for one week and aortic specimens were obtained for HE staining. Besides, thirty-two male ApoE-/-mice （10 weeks old） fed with normal food were selected as experimental group, which were randomly divided into 4 groups according to drug dosages （kilogram of body weight per day）, 50μg/（kg·d） group, 75μg/（kg·d） group, 100μg/（kg·d） group and blank control group with eight mice in each. Eight male C57BL/6J mice （10 weeks old） were selected as negative con-trol group. 3 weeks after medication, mice were sacrificed and aortic specimens were obtained for HE staining. More-over, The concentration of IL-10, IL-12 in serum were detected. Result 1. There were statistically significant differ-ence between the model group [（5.250±0.500） points] and the control group [（0.500±0.577） points] in aortic HE stain-ing and Roberts ＆amp;Thompson Scores in eleven weeks old mice. 2. After treated with different concentrations of trip-tolide for three weeks, the aortic HE staining and Roberts＆amp;Thompson scores of mice of those groups were in a de-scending order, which were （6.500 ± 0.189） points （blank control group）, （4.625 ± 0.183） points [100μg/（kg·d） group], （3.375±0.183） points [50μg/（kg·d） group] （1.375±0.183） points [75μg/（kg·d） group] and （0.000±0.000） points （nega-tive control group） with statistically significant differences among those groups. 3. The increase of serum IL-10 in the 75 μg/（kg·d） group was the most obvious one than those in other groups （P〈0.001）. Compared with blank control group, the concentration of serum IL-12 in other groups were lower and that in the 75μg/（kg·d） group was the low-est. The differences were statistically significant （P〈0.001）. Conclusion 1. AS model could be generated with eleven weeks old ApoE-/-mice. 2. ApoE-/-mice AS lesions had varying degrees of relief when treated with different concen-trations of triptolide for three weeks and the effect in the 75 μg/（kg·d） group was the best. 3. Triptolide could increase the expression level of IL-10, decrease the expression level of IL-12, inhibit the inflammatory response and reduce the formation of plaque, which might be one of the mechanisms of anti-atherosclerosis with immunosuppressant.