冠心康对载脂蛋白E基因敲除动脉粥样硬化小鼠血脂及炎性标志物的影响

目的：观察中药复方冠心康对载脂蛋白E（apolipoprotein E,ApoE）基因敲除（ApoE-/-）动脉粥样硬化小鼠血脂及炎性标志物C-反应蛋白（C-reactive protein,CRP）、血清淀粉样蛋白A（serum amyloid Aprotein,SAA）、纤维蛋白原（fibrinogen,Fbg）的影响,探讨中药干预动脉粥样硬化形成的可能机制。方法：70只ApoE-/-小鼠予高脂饲料喂养建立实验性动脉粥样硬化模型。14只C57BL/6J小鼠为正常对照组,给予普通饲料。70只ApoE-/-小鼠给予高脂饲料造模,于第12周龄时随机抽取正常组小鼠和模型组小鼠各2只,取主动脉苏木精-伊红染色检测易损斑块是否形成。68只ApoE-/-小鼠随机分为模型组、高剂量冠心康组（生药浓度为3.456g/mL）、中剂量冠心康组（1.728g/mL）、低剂量冠心康组（0.864g/mL）、阳性药物辛伐他汀组3mg/（kg.d）],每天1次灌胃治疗。连续灌胃8周后,经眼眶采血,分离血清。酶偶联比色法测定血清总胆固醇（total cholesterol,TC）及三酰甘油（triacylglycerol,TAG）含量,选择性沉淀法测定低密度脂蛋白胆固醇（low-density lipoproteincholesterol,LDL-C）及高密度脂蛋白胆固醇（high-densitylipoproteincholesterol,HDL-C）含量,比浊法测定载脂蛋白AⅠ（apolipoprotein AⅠ,ApoAⅠ）及载脂蛋白B（apolipoprotein B,ApoB）含量,双抗体夹心酶联免疫吸附测定（ABC）法检测ApoE、CRP、SAA和Fbg含量。结果：与模型组比较,各给药组血清TC、TAG、LDL-C、ApoB、CRP、SAA和Fbg含量下降（P〈0.05）,ApoAⅠ和HDL-C含量升高（P〈0.05）。高剂量冠心康组与低剂量冠心康组比较,血清HDL-C和ApoAⅠ含量升高（P〈0.05）,TAG、TC、ApoB、CRP、SAA和Fbg含量下降（P〈0.05）。与辛伐他汀组比较,髙剂量冠心康组血清HDL-C和ApoAⅠ含量升高（P〈0.05）,TAG、TC、ApoB、SAA和Fbg含量下降（P〈0.05）,中剂量冠心康组血清HDL-C含量上升（P〈0.05）,TC、ApoB和Fbg含量下降（P〈0.05）,低剂量冠心康组HDL-C和CRP含量升高（P〈0.05）,ApoB含量下降（P〈0.05）。结论：复方冠心康可改善ApoE基因敲除小鼠的脂质代谢紊乱,降低小鼠血清炎症标志物CRP、SAA和Fbg水平,抑制炎症反应进展,且以高剂量作用为突出,对小鼠动脉粥样硬化有一定的防治作用。

Effects of Chinese herbal medicine Guanxinkang on lipid metabolism and serum C-reactive protein, amyloid A protein and fibrinogen in apolipoprotein E-knockout mice with atherosclerosis

Objective：To observe the effects of Guanxinkang（GXK）decoction,a compound traditional Chinese herbal medicine,on serum lipids and apolipoprotein AⅠ（ApoAⅠ）,apolipoprotein B（ApoB）,apolipoprotein E（ApoE）,C-reactive protein（CRP）,serum amyloid A protein（SAA）and fibrinogen（Fbg）concentrations of ApoE-knockout mice with atherosclerosis,and to explore the mechanism of GXK decoction in anti-atherosclerosis.Methods：Seventy 6-week-old ApoE-knockout mice receiving a high-cholesterol diet were used to induce atherosclerosis and were randomly divided into 5 groups：untreated group,simvastatin group and low-（drug concentration is 0.864 g/mL）,medium-（1.728 g/mL）,and high-dose（3.456 g/mL）GXK groups.Another fourteen 6-week-old C57BL/6J mice were used as the normal control.Two 12-week-old mice were randomly selected from the normal control and the ApoE-knockout mice respectively to observe vulnerable plaque in the mouse＇s aortic by hematoxylin-eosin staining.Blood was collected from venous plexus of eye socket after gavage of corresponding drugs once daily for 8 weeks continuously,and then the serum was separated.Triglyceride（TAG）and total cholesterol（TC）were measured by enzyme-coupled assay;low-density lipoprotein cholesterol（LDL-C）and high-density lipoprotein cholesterol（HDL-C）were measured by selective precipitation method.Serum levels of ApoAⅠ and ApoB were determined by turbidimetry.Double-antibody sandwich enzyme-linked immunosorbent assay was used to detect ApoE,CRP,SAA and Fbg concentrations in serum.Results：Compared with the normal control group,the levels of serum TC,TAG,LDL-C,ApoB,CRP,SAA and Fbg in the untreated group were increased（P0.05）,and the serum concentrations of HDL-C,ApoA Ⅰ and ApoE in the untreated group were decreased（P0.05）.After treatment,GXK decoction and simvastatin improved the dyslipidemia by increasing the concentrations of ApoAⅠ and HDL-C and decreasing the concentrations of TC,TAG,LDL-C,ApoB,CRP,SAA and Fbg（P0.05）.The high-dose GXK decoction had the most marked effects on SAA and Fbg and the serum lipids compared with the low-dose and medium-dose GXK and simvastatin.Conclusion：GXK decoction may not only provide an active effect on hyperlipidemia,but also down-regulate the levels of serum CRP,SAA and Fbg.GXK decoction exerts an anti-atherosclerosis effect in ApoE-knockout mice.