广东药学院 硕士学位论文 毛茛总苷对大鼠血压、血流动力学影响及机制研究 姓名：杨智承 申请学位级别：硕士 专业：药物化学 指导教师：谭毓治 20100529 广东药学院硕士研究生学位论文 I 摘 要 本论文正文由三部分组成，第一部分主要介绍了毛茛总苷对正常大鼠血压和小鼠 耐缺氧的影响。第二部分介绍了毛茛总苷对麻醉大鼠血流动力学的影响。第三部分介 绍了毛茛总苷对大鼠血管平滑肌细胞内钙离子的影响。 第一部分 毛茛总苷对正常大鼠血压及小鼠耐缺氧的影响 第一部分 毛茛总苷对正常大鼠血压及小鼠耐缺氧的影响 本实验采用腹腔注射给药，股动脉插管、右颈总动脉心室插管和连续记录肢体 导联心电图的方法观察药物对麻醉大鼠心率(HR) 、收缩压(SP) 、舒张压(DP) 、平 均动脉血压(AP)的变化。KM 小鼠随机分为5 组，各组小鼠连续给药3 天，末次给药 后30 min 进行常压耐缺氧实验， 观察小鼠的存活时间。 结果显示： 毛茛总苷 120 mg/kg 和60 mg/kg 均能显著地减慢大鼠心率，显著降低SP、DP、AP (P 0.05或P 0.01)； TGRJ 各剂量组均能通过降低心脏后负荷，减慢心率，减少心脏做功显著性的降低大 鼠的心肌耗氧，对大鼠心电图无明显影响。与生理盐水组比，120 mg/kg 的毛茛总苷 能延长小鼠存活时间(P 0.05)，60 mg/kg 的毛茛总苷虽然能延长小鼠存活时间， 但差异无显著性意义。结果提示：毛茛总苷有减慢大鼠心率，降低动脉血压，降低大 鼠心肌耗氧的作用。120 mg/kg 的毛茛总苷能明显提高正常小鼠常压耐缺氧能力，延 长小鼠存活时间。 第二部分 毛茛总苷对大鼠血流动力学的影响 第二部分 毛茛总苷对大鼠血流动力学的影响 本实验采用股静脉注射方式给药，股动脉插管、右颈总动脉心室插管和连续记录 肢体导联心电图的方法观察药物对麻醉大鼠心率(HR) 、收缩压(SP) 、舒张压 (DP) 、平均动脉血压(AP) 、左心室收缩压(LVSP) 、左心室舒张压(LVDP) 、左心室 终末舒张压(LVEDP) 、左心室内压最大上升速率( + dp/dtmax) 、左心室内压最大下降 速率( - dp/dtmax) 和等容舒张时间室内压下降的时间常数(T) 的变化。结果显示：静 脉注射毛茛总苷 80 mg/kg 和40 mg/kg 后，均能显著地减慢大鼠心率，显著降低SP、 DP、AP、LVSP、- dp/dtmax和T值(P 0.05或P 0.05)。结果提示：毛茛总苷有减慢大鼠心率，降低动脉血压，抑制心肌 收缩和舒张功能的作用。 广东药学院硕士研究生学位论文 II 第三部分 毛茛总苷对大鼠血管平滑肌细胞内钙离子的影响 第三部分 毛茛总苷对大鼠血管平滑肌细胞内钙离子的影响 本实验采用新一代 Ca 2+ 荧光探针 Fluo-3/AM 标记培养的血管平滑肌细胞，荧光 分光光度计法检测细胞胞内钙荧光信号，观察毛茛苷对培养的大鼠血管平滑肌细胞胞 内 Ca 2+ 水平的影响。结果显示，去甲肾上腺素能显著性地促进胞外钙离子内流，而毛 茛总苷各剂量组均能降低由去甲肾上腺素引起的细胞内钙离子浓度升高，并呈一定的 量效关系。这提示毛茛总苷能抑制去甲肾上腺素引起的细胞内钙离子浓度增加。 关键词关键词：毛茛总苷；血压；血流动力学；血管平滑肌细胞；钙离子 广东药学院硕士研究生学位论文 i Efects and Mechanisms of Total Glycosides of Ranunculus japonicus on Blood Pressure，Hymodynamics in Rats Master Candidate: Zhicheng Yang Majior：phamaceutical chemistry Supervisor: Pro. Yuzhi Tan Abstract This thesis is composed of three parts. Partmainly introduced the effects of Total Glycosides of Ranunculus japonicus (TGRJ) on the BP in anesthetic rats and hypoxia tolerance in mice. Partcovered my study on the effects of TGRJ on the hemodynamics in anesthetic rats. Part reported the effect of TGRJ on intracellular calcium concentration in thoracic aortic vascular smooth muscle cells of rats. Part Effects of TGRJ on BP in anesthetic rats and hypoxia tolerance in mice In this thesis, changes of heart rate (HR), systolic arterial pressure (SP), diastolic arterial pressure (DP), average arterial pressure (AP) and consecutive electrocardiographic were recorded by cannulating femoral artery, right common carotid artery. KM mice were randomized into five groups, each group given orally with medicine continuously for three days. 30 minutes after the last administration, the survival time of the hypoxia tolerance mice under normal pressure were measured. Results showed that TGRJ 120 mg/kg and TGRJ 60 mg/kg can significantly decrease HR, SP, DP, AP. Different doses of TGRJ significantly reduce the oxygen consumption of cardiac muscle in rats by reducing after load, heart rate and cardio working. 120 mg/kg TGRJ can obviously prolong the survival time of mice under the normal pressure (P 0.05). And the results concluded that TGRJ reduce HR, arterial blood pressure and oxygen consumption of cardiac muscle in rats. TGRJ 120 mg/kg prolong the survival time of mice; enhance the hypoxia tolerance of mice under normal pressure. 广东药学院硕士研究生学位论文 ii Part Effects of TGRJ on the hemodynamics in rats In this thesis, changes of hemodynamic parameters including heart rate (HR) , systolic arterial pressure (SP) , diastolic arterial pressure (DP) , average arterial pressure (AP ), left ventricular pressure (LVSP) , left ventricular end-diastolic pressure (LVEDP) , left ventricular pressure development during isovolumetric contraction/relaxation (dp/dt max), and T were recorded by cannulating femoral artery, right common carotid artery and consecutive electrocardiographic. Results showed that TGRJ 80 mg/kg and TGRJ 40 mg/kg can significantly decrease HR, SP, DP, AP, LVSP, - dp/dt max and T (P 0.05 or P 0.01) in anesthetized rats, but there is no obvious change in LVEDP and + dp/dt max. And the results concluded that TGRJ reduces HR and BP. It has depressive effects on both myocardial contraction and diastole and negative effects on cardiac function in anesthetized rats. Part Effects of TGRJ on intercellular calcium in rat vascular smooth muscle cells Vascular smooth muscle cells were incubated with the calcium ion-sensitive fluorescent indicator fluo-3/AM, and then a fluorescence spectrophotometer was applied to measure changs of fluorescence intensity to investigate the effects of TGRJ on intracellular Ca 2+ concentration in VSMC of rats. Results showed that norepinephrine could significantly promote the Ca2+ into intracellular, and different dose of TGRJ could lower the increasing intracellular Ca2+ concentration induced by norepinephrine with a maximal respone in some dose-dependent relation, espescially at the concentration of 400g/ml. Keywords：total glycosides of Ranunculus japonicus，blood p