温阳法、滋阴法、活血法及复合法对肾阳虚型心力衰竭大鼠心室重构及心功能的影响

Effects of Different Chinese Medicine Therapeutic Methods on the Ventricular Remodeling and the Heart Function in Chronic Heart Failure Rats of Shen-yang Deficiency Syndrome

目的比较温阳法、滋阴法、活血法及复合法对肾阳虚型心力衰竭大鼠心室重构(ventricular re-modeling,VR)及心功能的影响。方法采用切除双侧甲状腺组织后持续静脉滴注阿霉素的方法制备心肾阳虚型心力衰竭大鼠模型。随机分为模型组生理盐水6mL/(kg·d)灌胃、温阳组温阳健心灵浸膏6mL/(kg·d)灌胃、活血组川芎丹参浸膏6mL/(kg·d)灌胃、滋阴组麦冬知母浸膏6mL/(kg·d)灌胃及复合组阴阳双补健心浸膏6mL/(kg·d)灌胃,每组10只。另选取10只大鼠作为正常对照组(正常组),自由饮食饮水。各组连续干预4周。观察大鼠左心室收缩压(left ventricular systolic pressure,LVSP)、左心室舒张末压(left ventricular end diastolic pressure,LVEDP)、左心室内压最大上升和下降速率maximal rate of left ventricular pressure of development(+dp/dtmax) and maximal rate of left ventricular pressure of decline(-dp/dtmax),测量动脉收缩压(systolic blood pressure,SBP)及舒张压(diastolic blood pressure,DBP),计算平均动脉压(mean arterial pressure,MAP),记录心率(heart rate,HR)。称取心脏质量(heart mass,HM)、左心室(包含室间隔)质量(left ventricular mass,LVM)及右心室质量(right ventricular mass,RVM),计算心脏质量指数(heart mass index,HMI)和左心室质量指数(left ventricular mass index,LVMI)。采用RT-PCR法测定大鼠心肌基质金属蛋白酶9(Matrix metalloproteinase-9,MMP-9)、组织金属蛋白酶抑制剂-1(tissue inhibitors of matrixmetalloproteinase-1,TIMP-1)mRNA表达。采用双抗体夹心ELISA法测定血清MMP-9、TIMP-1、N末端脑钠肽(N-terminal pro-brain natriuretic peptide,NT-proBNP)的含量。结果与正常组比较,模型组HR、LVEDP、HM、LVM、HMI、RVM、LVMI、NT-proBNP、MMP-9含量及MMP-9mRNA升高,SBP、DBP、MAP、LVSP、±dp/dtmax、TIMP-1含量及TIMP-1mRNA降低,差异均有统计学意义(P<0.05)。与模型组比较,温阳组SBP、DBP及±dp/dtmax升高,LVEDP、NT-proBNP、MMP-9降低;活血组HR、LVEDP、HM、LVM、HMI、RVM、LVMI、MMP-9mRNA、NT-proBNP、MMP-9含量均降低,±dp/dtmax、TIMP-1mRNA升高;复合组HR、LVEDP、±dp/dtmax、LVMI、NT-proBNP、MMP-9降低,TIMP-1升高;差异均有统计学意义(P<0.05)。与温阳组比较,滋阴组SBP及+dp/dtmax降低,活血组HR、MMP-9mRNA降低,复合组HR降低,差异均有统计学意义(P<0.05)。其余各指标组间比较,差异无统计学意义(P>0.05)。结论活血法及复合法调节肾阳虚型心力衰竭大鼠MMP-9、TIMP-1表达,有效减轻VR,改善心功能。

Objective To observe the effects of warming yang method, nourishing yin method, activating blood method, and the combined treatment method on the ventricular remodeling （~ VR） and the heart function of heart failure （HF） rats of Shen-yang deficiency syndrome （SYDS）. Methods The Sprague-Dawley （SD） HF rat model of SYDS was established by continuously intravenous dripping adriamycin after ectomizing bilateral thy- roid tissues. Rats were then randomly divided into the model group （administered with normal saline at the daily dose of 6 mL/kg by gastrogavage）, the warming yang group （administered with Wenyang Jianxinling Extractum at the daily dose of 6 mL/kg by gastrogavage）, the activating blood group （administered with Ligusticum Wallichfi and Salvia Miltiorrhiza Extractum at the daily dose of 6 mL/kg by gastrogavage）, the nourishing yin group （ ad- ministered with Radix Ophiopogonis and Rhizoma Anemarrhenae Extractum at the daily dose of 6 mL/kg by gas- trogavage）, and the combined treatment group （ administered with Yin-Yang Supplementing Extractum at the dai- ly dose of 6 mL/kg by gastrogavage）, 10 in each group. Another 10 SD rats were taken as the normal control group. They ate food and drank water freely. All rats were intervened for four successive weeks. The left ventric- ular systolic pressure （LVSP）, left ventricular end diastolic pressure （LVEDP）, maximal rate of left ventricular pressure of development （ ＋dp/dtmax）, and maximal rate of left ventricular pressure of decline （-dp/dtmax） were observed. The systolic blood pressure （SBP） and the diastolic blood pressure （DBP） were determined. The mean arterial pressure （MAP） was calculated. The heart rate （HR） was recorded. Then all rats were killed and their hearts were taken out to weigh the heart mass （ HM）, the left ventricular mass （ LVM）, the right ventric- ular mass （RVM）. The heart mass index （HMI） and the left ventricular mass index （LVMI） were calculated. The mRNA expression of matrix metalloproteinase-9 （MMP-9） and tissue inhibitor of metalloproteinase-1 （TIMP1） in the myocardium were detected using RT-PCR. The serum levels of N-terminal pro-brain natriuretic peptide （NT-proBNP）, MMP-9, and TIMP-1 were assayed by double antibody sandwich ELISA. Results Com- pared with the normal control group, HR, LVEDP, HM, LVM, HMI, RVM, LVMI, NT-proBNP, MMP-9, and MMP-9 mRNA significantly increased, but SBP, DBP, MAP, LVSP, _dp/dtmax, TIMP-1, and TIMP-1 mRNA significantly decreased in the model group with statistical difference （P 〈 0.05）. Compared with the model group, SBP and DBP, _ dp/dtmax increased,while LVEDP, NT-proBNP, and MMP-9 decreased in the warming yang group. HR, LVEDP, HM, LVM, HMI, RVM, LVMI, MMP-9 mRNA, NT-proBNP, and MMP-9 significantly de- creased, while TIMP-1 mRNA increased in the activating blood group. HR, LVEDP, 4-dp/dtmax, LVMI, NT- proBNP, and MMP-9 decreased,while TIMP-1 increased in the combined treatment group, showing statistical difference （P〈0.05）. Compared with the warming yang group, SBP and ＋dp/dtmax decreased in the nouris- hing yin group; HR and MMP-9 mRNA decreased in the activating blood group, HR decreased in the combined treatment group, all showing statistical difference （P 〈0.05）. There was no statistical difference in the rest indi- ces （ P 〉0.05）. Conclusien Activating blood method and combined treatment method could regulate the expres- sions of MMP-9 and TIMP1 mRNA of HF rats of SYDS, effectively ameliorate the VR, and improve the HF.