摘要
目的 探讨绞股蓝总皂苷对动脉粥样硬化大鼠主动脉炎症反应的影响及预测部分绞股蓝有效成分.方法 将60只SD大鼠分为空白对照组(Nor组,)、模型组(Mod组)、辛伐他汀组(Sim组,5 mg/kg灌胃,13周)、绞股蓝总皂苷组(GPs组,160 mg/kg灌胃,13周)、GW9662组(PPAR-γ 抑制剂,10 mg/kg灌胃,13周)及GPs+GW9662组[GPs 160 mg/kg及GW966210 mg/(kg·d),灌胃,13周],其中Nor组普通饮食,其他组予以高脂饲料联合腹腔注射维生素D3以构建动脉粥样硬化大鼠模型;建模13周后取大鼠血液、胸主动脉和腹主动脉,HE切片染色观察胸主动脉组织学变化,ELISA测定血清中IL-6含量,Western blot检测腹主动脉过氧化酶体增殖物激活受体-γ(PPAR-γ),Wnt3a及β-catenin蛋白表达、qRT-PCR检测腹主动脉PPAR-γ及Wnt3a及β-catenin mRNA表达;利用分子对接初步筛选绞股蓝总皂苷与PPAR-γ 亲和力较高的有效成分.结果 与Nor组比较,Mod组及其他干预组大鼠主动脉切片可见AS特征性病变,其中GW9662组病变明显,辛伐他汀和绞股蓝总皂苷各干预组病变较轻;与Mor组相比,GPs组大鼠血清白细胞介素-6(IL-6)含量显著减少;与Mor组大鼠腹主动脉比较,GPs组PPAR-γ蛋白和mRNA的表达显著减少(P<0.05)、其他非阳性对照干预组均不同程度增高,而Mor组Wnt3a、β-catenin和mRNA表达与Nor组相比明显升高(P<0.05)、其他非阳性对照干预组均不同程度减少,其中GPs组显著降低;分子对接结果显示野黄芩苷、胆固醇、菜油甾醇、谷甾醇、3'-甲基鼠李素对PPAR-γ具有较强的亲和力.结论 绞股蓝总皂苷可能是通过调节PPAR-γ 与Wnt经典途径改善大鼠动脉粥样硬化,野黄芩苷、胆固醇、菜油甾醇、谷甾醇、3'-甲基鼠李素成分是绞股蓝部分有效成分.
Abstract
Objective To investigate the effect of gypenosides on the inflammatory response of aorta in rats with atherosclerosis ( AS ) and to predict the active components of gypenosides ( GPs ) . Methods Sixty SD rats were divided into the blank control group (Nor group),model group (Mod group),simvastatin group ( Sim group,with 5 mg/kg gavage for 13 weeks),gypenosides group ( GPs group,with 160 mg/kg gavage for 13 weeks),GW9662 group (with PPAR-γ inhibitor,10 mg/kg gavage for 13 weeks),and GPs+GW9662 group[GPs 160 mg/kg and GW966210 mg/( kg·d) by gavage for 13 weeks],in which the Nor group was fed normal diet,and the other groups were fed high-fat diet combined with intraperitoneal injection of vitamin D3 to construct an AS rat model;blood,thoracic aorta,and abdominal aorta of the rats were collected 13 weeks after modeling,and the thoracic aorta was stained with HE section to observe the histological changes. Thirteen weeks after modelling,the blood and abdominal aorta were stained to observe the histological changes in the thoracic aorta,the serum IL-6 content was determined by ELISA,the protein expressions of PPAR-γ,Wnt3a,and β-catenin were detected by Western blot,and the expressions of PPAR-γ,Wnt3a,and β-catenin mRNA in the abdominal aorta were detected by qRT-PCR;and the molecular docking was used to filter out active ingredients with higher affinity for GPs and PPAR-γ. Results Compared with the Nor group,the aortic sections of rats in the Mod group and other intervention groups showed characteristic lesions of AS,among which the lesions in the GW9662 group were obvious and those in the simvastatin and GPs intervention groups were milder;compared with the Mor group,the serum IL-6 content of the rats in the GPs group was significantly reduced;compared with the abdominal aorta of the rats in Mor group,the expression of PPAR-γ protein and mRNA was significantly reduced ( P<0.01) in the GPs group.The expression of PPAR-γ protein and mRNA was significantly reduced in the GPs group ( P<0.05 ),and increased to different degrees in the other non-positive control intervention groups,whereas the expression of Wnt3a,β-catenin,and mRNA was significantly elevated in Mor group compared with that of Nor group ( P<0 .05 ),and reduced to different degrees in the other non-positive control intervention groups,which was significantly reduced in GPs group. The molecular docking results showed that baicalin,cholesterol,canola sterol,sitosterol and 3'-methyl rhamnetin had a strong affinity for PPAR-γ. Conclusion Gypenosides may improve AS in rats by regulating PPAR-γand Wnt classical pathway,and baicalin,cholesterol,canola sterol,sitosterol and 3'-methyl rhamnetin are some of the active ingredients of GPs.
基金项目
国家自然科学基金资助项目(82060776)
贵州省科技计划项目(黔科合基础[2020]1Y388)
国家本科创新创业人才培养计划项目(202010660028)
国家本科创新创业人才培养计划项目(202010660005)
国家本科创新创业人才培养计划项目(201910660037)
维普
万方数据