基质金属蛋白酶9通过上调血管紧张素Ⅱ加剧其致心室重构作用  

Matrix metalloproteinase-9 aggravates ventricular remodeling by up-regulating angiotensinⅡ

作　　者：骆露媛 古思奇 周万兴[1] 陈立芝 游昕超 李瑞莹[4] 张卫[1] 宋玉兰 LUO Lu-yuan;GU Si-qi;ZHOU Wan-xing;CHEN Li-zhi;YOU Xin-chao;LI Rui-ying;ZHANG Wei;SONG Yu-lan(Department of Internal Cardiology,The First Affiliated Hospital of GuangDong Pharmaceutical University,Guangzhou 510080,China)

机构地区：[1]广东药科大学附属第一医院心血管内科,广东广州510080 [2]惠州市第三人民医院心血管内科 [3]广东药科大学附属第一医院动物中心,广东广州510080 [4]广东药科大学附属第一医院检验科,广东广州510080 [5]广东药科大学附属第一医院病理科,广东广州510080

出　　处：《中华高血压杂志》2019年第9期859-866,共8页Chinese Journal of Hypertension

基　　金：广东省科技计划项目(2017ZC0204)

摘　　要：目的探讨血管紧张素Ⅱ(AngⅡ)是否介导基质金属蛋白酶9(MMP-9)的致心室重构作用及其机制,为进一步阐明心室重构发病机制和寻找防治心室重构新靶点提供证据。方法 36只8周龄雄性Wistar大鼠随机分为对照组(n=12)、MMP-9组(n=12)、MMP-9+奥美沙坦组(n=12),分别给予生理盐水、重组MMP-9生理盐水溶液(2.1ng/g)、重组MMP-9生理盐水溶液(2.1ng/g)各2mL腹腔注射,每周2次;同时,MMP-9+奥美沙坦组加奥美沙坦(3mg/kg)1mL,其余两组各加1mL生理盐水灌胃,1次/d。第28天,每组随机抽取6只大鼠作为4周阶段实验组,各组剩余大鼠作为8周阶段实验组。实验结束时超声心动图检查,测量左心室质量指数(LVMI),心肌组织切片苏木精-伊红染色法(HE)和Van Gieson(VG)染色观察心肌细胞和胶原分布情况。酶联免疫吸附试验(ELISA)法测定血清血管紧张素Ⅰ(AngⅠ)、AngⅡ、血管紧张素转换酶(ACE)、MMP-9水平。逆转录聚合酶链反应(RT-PCR)法测血和心肌组织MMP-9mRNA表达。免疫印迹法(Western blot)测心肌组织MMP-9蛋白表达。结果在实验第4周和第8周,与对照组比,MMP-9组心肌组织MMP-9mRNA和蛋白表达水平升高,血清MMP-9[4周:(80.66±4.73)比(48.54±6.94);8周:(143.30±37.44)比(48.10±7.96)μg/L]、AngⅠ、AngⅡ[4周:(101.82±13.57)比(55.07±7.06);8周:(100.32±11.65)比(66.09±8.50)μg/L]、ACE、LVMI均增高,胶原容积分数(CVF)降低(P<0.05)。经奥美沙坦干预后,第4和第8周心肌MMP-9mRNA和蛋白表达水平均较MMP-9组降低,血清MMP-9[4周:(56.76±5.96)比(80.66±4.73),8周:(83.25±16.98)比(143.30±37.44)μg/L]、AngⅡ[(69.31±6.58)比(101.82±13.57),8周:(74.77±18.08)比(100.32±11.65)μg/L],LVMI均比MMP-9组降低;CVF回升(均P<0.05),心肌形态学明显改善,8周组比4周组更加明显;而AngⅠ、ACE变化无统计学意义(P>0.05)。心脏超声仅在8周的MMP-9组发现左心室舒张末期内径、左心室舒张末期容积较对照组增高(P<0.05),经奥美沙坦干�Objective To explore whether angiotensinⅡ(AngⅡ)mediates the effect of matrix metalloproteinase-9(MMP-9)on ventricular remodeling and its mechanism,so as to provide evidence for further elucidation of the pathogenesis of ventricular remodeling and search for new targets for prevention and treatment of ventricular remodeling.Methods Thirty-six eight-week male Wistar rats were randomly divided into control group(n=12),MMP-9 group(n=12)and MMP-9+olmesartan group(n=12),which were given normal saline,recombinant MMP-9 normal saline solution(2.1 ng/g),recombinant MMP-9 normal saline solution(2.1 ng/g)by each 2 mL intraperitoneal injection,twice a week respectively.In addition,the olmesartan group was given 1 mL olmesartan solution(3 mg/kg),and the others were given normal saline 1 mL by gavage once a day.On the 28 th day,6 rats were randomly selected from each group as the 4-week phase experimental group,and the remaining rats in each group as the 8-week experimental group.At the end of the experiment,echocardiography and myocardial section microscopy examination(HE and VG stain)were performed.The mRNA and protein levels of MMP-9 in myocardial tissue were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot.The levels of angiotensinⅠ(AngⅠ),angiotensinⅡ(AngⅡ),angiotensin converting enzyme(ACE)and MMP-9 in serum were detected by enzymelinked immunosorbent assay(ELISA).Left ventricular mass index(LVMI)was measured and calculated,and the collagen volume fraction(CVF)was obtain by image analysis.Results At the fourth and eighth week of the experiment,compared with the control group,the mRNA and protein levels of MMP-9 in myocardial tissue,and the level of MMP-9[4 week:(80.66±4.73)vs(48.54±6.94),8 week:(143.30±37.44)vs(48.10±7.96)μg/L],AngⅠ,AngⅡ[4 week:(101.82±13.57)vs(55.07±7.06),8 week:(100.32±11.65)vs(66.09±8.50)μg/L]and ACE in serum and LVMI were significantly higher;while CVF was lower significantly in the MMP-9 group(all P<0.05).After the intervention of olmesarta

关 键 词：基质金属蛋白酶9 血管紧张素Ⅱ 大鼠 心室重构 奥美沙坦 

分 类 号：R54[医药卫生—心血管疾病]