桃仁红花煎通过抑制淋巴管增生改善ApoE^-/-小鼠动脉粥样硬化的炎症反应

TaorenHonghua formula inhibits lymphangiogenesis to improve inflammatory response of atherosclerosis in ApoE^-/-Mice

目的:观察桃仁红花煎对ApoE^-/-基因敲除鼠动脉粥样硬化模型的心脏淋巴管、纵膈淋巴结T细胞含量以及相关炎症因子指标的影响.方法:将ApoE^-/-基因敲除鼠随机分为空白组、模型组、桃仁红花煎高剂量组、中剂量组和低剂量组,模型组和给药组采用高脂饮食造模8周,桃仁红花煎通过灌胃给药,空白组采用质量分数为0.9%氯化钠溶液灌胃作为对照.给药周期为12周,频率为每天2次,每次灌胃0.3 mL.采用冰冻切片免疫荧光法检测淋巴管及淋巴结中T细胞含量,油红O染色法评估主动脉窦的斑块面积,提取主动脉总RNA并反转录后,采用实时荧光定量PCR法分析相关基因:白细胞介素-6(IL-6)、白细胞介-10(IL-10)和肿瘤坏死因子-α(TNF-α)的mRNA相对表达水平,炎症因子采用ELISA方法进行检测.结果:与模型组比较,桃仁红花煎可显著降低小鼠体质量、甘油三酯和总胆固醇浓度(P<0.05,P<0.01),减小主动脉窦斑块面积,抑制心脏淋巴管增生,减少淋巴结中T细胞含量,降低IL-6和TNF-α的质量浓度以及mRNA相对表达量(P<0.01).结论:桃仁红花煎可有效抗动脉粥样硬化,其机制可能与减少心脏淋巴管数目、淋巴结中T细胞含量以及炎症因子表达相关.

Objective:To investigate the efficacy of TaorenHonghua Jian on the content of heart lymphatic vessels,T lymphocytes in the mediastinal lymph nodes and related inflammatory factors in ApoE^-/-atherosclerosis model.Methods:ApoE^-/-mice were randomly divided into blank group,model group,high dose group,medium dose group and low dose group.Model group and drug groups were induced through high fat diet for 8 weeks.TaorenHonghua Jian was gastrointestinal administrated and the blank group was administrated by the same amount with 0.9%sodium chloride solution as control group.The intervention period was 12 weeks,the frequency was twice a day,and the amount of gastric perfusion was 0.3 mL.Heart lymphatic vessels and T cell contents in lymph nodes were detected by frozen section immunofluorescence method.The plaque areas of aortic sinus were evaluated by oil red O staining method.Total RNA of aorta was extracted and reverse transcribed.Real-time quantitative PCR was used to analyze the related genes(IL-6,IL-10 and TNF-α).The relative expression levels of inflammatory cytokines were detected by ELISA.Results:Compared with the model group,TaorenHonghua Jian could significantly reduce the body weight,triglycerides and total cholesterol(P<0.05,P<0.01),small the areas of aortic sinus plaque,inhibit heart lymphangiogenesis,reduce T cell contents in lymph nodes,reduce the production and mRNA relative expression of IL-6 and TNF-α(P<0.01).Conclusion:TaorenHonghua Jian effectively against atherosclerosis.Its mechanism might be related to reducing the numbers of heart lymphatic vessels and T cells of lymph nodes,and expression of inflammatory factors.