复方七芍降压片通过调控MMP-2/TIMP-2对冠心病大鼠血管重塑及动态血压的影响

Effect of Compound Qishao Jiangya Tablet on Vascular Remodeling and Ambulatory Blood Pressure in Rats with Coronary Heart Disease by Regulating MMP-2/TIMP-2

目的探讨复方七芍降压片通过调控基质金属蛋白酶-2(MMP-2)/组织金属蛋白酶抑制物-2(TIMP-2)对冠心病大鼠血管重塑的作用及对动态血压的影响。方法60只大鼠,随机取10只作为空白对照组(A组),另外50只采用冠状动脉结扎法制备冠心病大鼠模型,造模成功44只,随机分为模型组(B组)、卡托普利组(C组)、复方七芍降压片低剂量组(D组)、复方七芍降压片高剂量组(E组),各11只。采用无创血压仪检测各组大鼠收缩压、舒张压;酶联免疫吸附法(ELISA)检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、丙二醛(MDA)、活性氧(ROS);苏木精-伊红(HE)染色法观察胸主动脉形态学改变,检测血管重塑参数中膜厚度(MT)、管腔内径(LD),计算MT/LD;蛋白质印迹法(Western Blot)检测MMP-2、TIMP-2蛋白相对表达量。结果治疗1周、2周、3周、4周时,与A组比较,B组收缩压及舒张压升高(P<0.05);与B组比较,C组、D组、E组收缩压及舒张压降低(P<0.05);与D组比较,C组、E组收缩压及舒张压降低(P<0.05)。与A组比较,B组IL-1β、IL-6、TNF-α、ROS、MDA升高(P<0.05);与B组比较,C组、D组、E组IL-1β、IL-6、TNF-α、ROS、MDA降低(P<0.05);与D组比较,C组、E组IL-1β、IL-6、TNF-α、ROS、MDA降低(P<0.05)。HE染色显示,D组内皮增生程度减轻,部分平滑肌细胞向内膜下增生迁移,弹性纤维稍增多,C组、E组管壁无明显增厚,可见轻度内皮增生增厚,中膜平滑肌层、弹性纤维无明显病理改变。与A组比较,B组MT、MT/LD、MMP-2蛋白相对表达量及MMP-2/TIMP-2比值升高(P<0.05);与B组比较,C组、D组、E组MT、MT/LD、MMP-2蛋白相对表达量及MMP-2/TIMP-2比值降低(P<0.05);与D组比较,C组、E组MT、MT/LD、MMP-2蛋白相对表达量及MMP-2/TIMP-2降低(P<0.05);与A组比较,B组LD、TIMP-2蛋白相对表达量降低(P<0.05);与B组比较,C组、D组、E组LD、TIMP-2蛋白相对表达量升高(P<0.05);与D组比较,C组、E组LD、TIMP-2蛋白相对表达量升高(P<0.05)。结论复方七芍降压片可改善冠心病大鼠血管重塑,稳定动态血压,减轻氧化应激、炎症反应,作用机制可能与抑制MMP-2/TIMP-2通路有关。

Objective To investigate the effect of Compound Qishao Jiangya Tablet on vascular remodeling and ambulatory blood pressure in rats with coronary heart disease by regulating matrix metalloproteinase-2(MMP-2)/tissue inhibitor of metalloproteinase-2(TIMP-2).Methods Sixty rats were selected,and 10 rats were randomly selected as blank control group(group A).The remaining 50 rats were prepared as rat models with CHD by coronary artery ligation,and 44 rats were successfully modeled.The CHD modle rats were randomly divided into model group(group B),captopril group(group C),low-dose Compound Qishao Jiangya Tablet group(group D),and high-dose Compound Qishao Jiangya Tablet group(group E),with 11 rats in each group.The systolic blood pressure and diastolic blood pressure were measured by noninvasive blood pressure meter.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),and reactive oxygen species(ROS)were detected by enzyme-linked immunosorbent assay(ELISA).The morphological changes of thoracic aorta were observed by hematoxylin eosin(HE)staining.The medial thickness(MT)and luminal diameter(LD)of vascular remodeling parameters were detected.The expressions of MMP-2 and TIMP-2 were detected by Western Blot.Results At 1,2,3,and 4 weeks of treatment,compared with group A,systolic blood pressure and diastolic blood pressure in group B increased(P<0.05).Compared with group B,systolic blood pressure and diastolic blood pressure of group C,group D,and group E decreased(P<0.05).Compared with group D,systolic blood pressure and diastolic blood pressure in group C and group E decreased(P<0.05).Compared with group A,IL-1β,IL-6,TNF-α,ROS,and MDA in group B increased(P<0.05).Compared with group B,IL-1β,IL-6,TNF-α,ROS,and MDA in group C,group D,and group E decreased(P<0.05).Compared with group D,IL-1β,IL-6,TNF-α,ROS and MDA in group C and group E decreased(P<0.05).HE staining showed that,the degree of endothelial proliferation was reduced,some smooth muscle cells migrated to subintimal hyperplasia,and elastic fibers increased slightly in group D In group C and group E,there was no obvious thickening of the vessel wall,but mild thickening of the endothelium was observed.No obvious pathological changes were found in the smooth muscle layer and elastic fibers of the media.Compared with group A,the relative expression of MT,MT/LD,MMP-2 protein,and MMP-2/TIMP-2 in group B increased(P<0.05)Compared with group B,the relative expression of MT,MT/LD,MMP-2 protein and MMP-2/TIMP-2 in group C,group D,and group E decreased(P<0.05).Compared with group D,the relative expression of MT,MT/LD,MMP-2 protein,and MMP-2/TIMP-2 in group C and group E decreased(P<0.05).Compared with group A,the relative expression of LD and TIMP-2 protein in group B decreased(P<0.05).Compared with group B,the relative expression of LD and TIMP-2 protein in group C,group D,and group E increased(P<0.05).Compared with group D,the relative expression of LD and TIMP-2 protein in group C and group E increased(P<0.05).Conclusion Compound Qishao Jiangya Tablet can improve vascular remodeling,stabilize ambulatory blood pressure,reduce oxidative stress and inflammatory reaction in rats with CHD.The mechanism may be related to the inhibition of MMP-2/TIMP-2 pathway.