三七总皂苷对载脂蛋白E基因敲除小鼠脾脏单个核细胞白细胞介素17A、白细胞介素10和肿瘤坏死因子α表达的影响

Effect of Total Panax Notoginsenosides on Expression of Interleukin-17A,Interleukin-10 and Tumor Necrosis Factor-α in Spleen Mononuclearcells of Apolipoprotein EKnockout Mice

目的研究三七总皂苷(TPNS)对动脉粥样硬化(As)小鼠脾脏单个核细胞白细胞介素17A(IL-17A)、肿瘤坏死因子α(TNF-α)和白细胞介素10(IL-10)表达的影响。方法载脂蛋白E基因敲除(Apo E-/-)小鼠高脂饮食喂养,小鼠20周龄时分为模型组及TPNS大、小剂量组。用药组分别给予TPNS 40 mg/kg、120 mg/kg灌胃8周。采用全自动生化分析仪检测各组小鼠血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDLC)浓度;HE染色观察斑块形态并计算斑块面积/管腔面积;ELISA法检测小鼠脾脏单个核细胞IL-17A、IL-10和TNF-α表达。结果 TPNS可以降低Apo E-/-小鼠的血清TC、TG、LDLC浓度,大剂量组TC、TG、LDLC浓度分别为16.4±2.2 mmol/L、1.36±0.23 mmol/L和5.9±1.2 mmol/L,模型组TC、TG、LDLC浓度分别为28.2±4.0 mmol/L、2.08±0.17 mmol/L和10.0±1.9 mmol/L,差异有显著性(P均<0.001)。TPNS有延缓As斑块形成的作用(模型组及大、小剂量TPNS组的斑块面积/管腔面积分别为31.3%±5.1%、14.1%±5.0%和24.2%±4.9%,P均<0.05)。大剂量TPNS可以提高小鼠脾脏单个核细胞的IL-10表达,与模型组相比差异有显著性(40.9±2.2比36.3±2.8,P<0.05),小剂量TPNS组略有升高趋势,但差异无统计学意义。TPNS可以降低小鼠脾脏单个核细胞IL-17A和TNF-α的表达,以大剂量组效果显著,差异有统计学意义(IL-17A 18.1±1.5比22.8±3.1,P<0.05;TNF-α18.3±1.2比22.9±0.7,P<0.001)。结论 TPNS可以降低Apo E-/-小鼠血清TC、TG、LDLC的水平,升高脾脏单个核细胞IL-10的表达,同时降低IL-17A和TNF-α的表达,这可能是其抗As的重要机制之一。

Aim To estimate the effect of total panax notoginsenosides（ TPNS） on expression of interleukin-17A（ IL-17A）,interleukin-10（ IL-10） and tumor necrosis factor-α（ TNF-α） in apolipoprotein E-knockout（ Apo E-/-） mice.Methods Apo E-/-mice were fed with a high fat,high cholesterol diet containing 15% fat and 0. 25% cholesterol. At 20 weeks of age,Apo E-/-mice were randomized into three groups,the model group,the TPNS low dose group and the TPNS high dose group. Mice in TPNS low dose group were given oral doses of TPNS at 40 mg/kg,once a day for 8 weeks. Mice in TPNS high dose group were given oral doses of TPNS at 120 mg/kg,once a day for 8 weeks. Mice in model group weregiven an equal volume of distilled water orally. After 8 weeks,blood and aortas were obtained. Serum levels of lipid were analyzed,ratio of plaque area to vessel area and the expression of IL-17 A,IL-10 and TNF-α were examined by histological staining and ELISA respectively. Results It was observed in our study that serum levels of lipid,ratio of plaque area to vessel area,and expressions of IL-17 A and TNF-α were lower in mice which were given TPNS. Serum total cholesterol,triglyceride and low density lipoprotein cholesterol in model group was 28. 2 ± 4. 0 mmol/L,2. 08 ± 0. 17 mmol/L and 10. 0 ± 1.9 mmol/L respectively,which were 16. 4 ± 2. 2 mmol/L,1. 36 ± 0. 23 mmol/L and 5. 9 ± 1. 2 mmol/L in TPNS high dose group（ P〈0. 001） and 20. 4 ± 1. 1 mmol/L（ P〈0. 05）,1. 91 ± 0. 25 mmol/L（ P〉0. 05） and 8. 0 ± 0. 8 mmol/L（ P〈0.05） in TPNS low dose group. The ratio of plaque area to vessel area was 31. 3% ± 5. 1% in model group,14. 1% ± 5. 0%in TPNS high dose group and 24. 2% ± 4. 9% in TPNS low dose group（ P〈0. 05）. High dose of TPNS could down-regulate the expressions of IL-17 A and TNF-α in spleen mononuclearcell（ IL-17 A 18. 1 ± 1. 5 vs 22. 8 ± 3. 1,P〈0. 05; TNF-α 18. 3 ±1. 2 vs 22. 9 ± 0. 7,P〈0. 001）. On the other hand,high dose of TPNS could up-regulate the expression of IL-10 in spleen mononuclearcell（ 40. 9 ± 2. 2 vs 36. 3 ± 2. 8,P〈0. 05）. Conclusion These results suggest that TPNS could prevent atherosclerosis by lowering serum lipid levels and regulating spleen mononuclearcell IL-17 A,IL-10 and TNF-α expression.TPNS may have implications for clinical treatment of atherosclerosis vascular disease.