基于“肠道菌群-炎症”通路探讨丹蒌片防治ApoE^-/-小鼠动脉粥样硬化的作用及机制

Based on“intestinal flora-inflammation”pathway to investigate effect and mechanism of Danlou Tablet on atherosclerosis of ApoE^-/-mice

目的观察丹蒌片对高脂喂养动脉粥样硬化模型ApoE^-/-小鼠的影响,从菌群失调诱发炎症角度探讨丹蒌片抗动脉粥样硬化的作用机制。方法采用C57BL/6J野生型小鼠作为对照,高脂喂养ApoE^-/-小鼠24周后随机分为模型组、丹蒌片组,连续ig给药8周后取血测定血脂水平;ELISA法检测血清中脂多糖(LPS)水平;动脉大体油红O及主动脉窦HE染色观察斑块形成;粪便16 SrRNA扩增子测序检测肠道菌群;GC-MS法检测短链脂肪酸;实时荧光定量PCR法检测炎症因子肿瘤坏死因子-α(TNF-α)、细胞间黏附分子-1(ICAM-1)和白细胞介素-1β(IL-1β)mRNA的表达。结果与对照组比较,模型组小鼠血脂水平升高,肠道菌群失调,有害菌增加,有益菌减少,血清LPS水平升高,主动脉周围炎症水平升高;与模型组比较,丹蒌片干预可明显降低小鼠总胆固醇(TC)、三酰甘油(TG)及低密度脂蛋白胆固醇(LDL-C)水平(P<0.01),有效减少斑块面积;调节肠道菌群,进而降低血清LPS及主动脉周围炎症因子TNF-α、ICAM-1和IL-1β水平(P<0.01)。结论丹蒌片通过改善菌群结构,降低菌群失调诱发的炎症反应发挥抗动脉粥样硬化作用。

Objective To investigate the efficacy and mechanism of Danlou Tablet against atherosclerosis model of ApoE^-/-mice fed with high fat diet.Methods C57 BL/6 J mice were used as controls and ApoE^-/-mice were randomly divided into two groups after 24 weeks of high fat feeding,including the model group received saline and the treatment group received Danlou Tablet.Animals were executed after 8 weeks of treatment and serum was collected to measure blood lipids;Plaque formation in the aorta was observed by red O and HE staining;16 S rRNA sequencing was used to analyze changes in intestinal flora,and GC-MS test for detection fecal SCFAs content,ELISA for the determination of serum LPS,and real time PCR for detection of mRNA expression.Results Compared with the control group,the blood lipid levels were increased;intestinal flora was imbalance with increased harmful bacteria and reduced beneficial bacteria;The level of serum LPS and inflammation around the aorta were increased in the model group.Compared with the model group,the contents of TG,TC,LDL-C and the plaque area of Danlou Tablet group were decreased(P<0.01);Danlou Tablet group can regulate intestinal flora,thus effectively reducing serum LPS and inflammatory factors TNF-α,ICAM-1 and IL-1βlevels around the aorta(P<0.01).Conclusion Danlou Tablet exerts an anti-atherosclerosis action with favorable efficacy through restructing the intestinal flora stucture,inhibiting endotoxin releasing and constraining the inflammatory response induced by dysbacteriosis.