醋柳黄酮及其单体抑制心肌肥厚的机理研究

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1、中图分类号：R89；R36；R39硕士学位论文醋柳黄酮及其单体抑制心肌肥厚的机理研究院（系）、所临床医学院研究生姓名宋鑫学科、专业内科学导师姓名刘应才教授二Ο一二年四月1目录醋柳黄酮及其单体抑制心肌肥厚的机理研究„„„„„„111.1中文摘要„„„„„„„„„„„„„„„„„„„„„31.2英文摘要„„„„„„„„„„„„„„„„„„„„„51.3前言„„„„„„„„„„„„„„„„„„„„„„„91.4材料与方法„„„„„„„„„„„„„„„„„„„„111.5结果„„„„„„„„„„„„„„„„„„

2、„„„„„231.6讨论„„„„„„„„„„„„„„„„„„„„„„„321.7结论„„„„„„„„„„„„„„„„„„„„„„„411.8参考文献„„„„„„„„„„„„„„„„„„„„„411.9英汉缩略词对照表„„„„„„„„„„„„„„„„„462致谢„„„„„„„„„„„„„„„„„„„„„„„47醋柳黄酮及其单体对心肌肥厚抑制作用的研究进展（综述）3„„„„„„„„„„„„„„„„„„„„„„„„„472醋柳黄酮及其单体抑制心肌肥厚的机理研究摘要目的：在培养原代大鼠心肌细胞的基础上，用血管紧

3、张素Ⅱ诱导新生大鼠心肌细胞肥大为模型，探讨醋柳黄酮及其单体（槲皮素与异鼠李素）抑制心肌肥厚的作用机理是否与钙调神经磷酸酶信号通路有关。方法：取新生SD大鼠心肌细胞(1-3d)，运用差速贴壁分离法提纯后进行培养。心肌细胞常规培养72小时，换无血清培养基同步化24小时后，分别加入不同药物进行孵育。实验分组：①对照组（Control）：不加任何处理因素，与加入处理因素的组别共同进行换液处理；②模型组（AngⅡ）：加入Ang-6Ⅱ10mol/L，建立心肌肥厚模型；③醋柳黄酮组（TFHgroup）：预先加入-6醋

4、柳黄酮100mg/L，再加入AngⅡ10mol/L;④槲皮素组(Quergroup)：预-6先加入槲皮素100umol/L，再加入AngⅡ10mol/L;⑤异鼠李素组(Isor-6group)：预先加入异鼠李素100umol/L，再加入AngⅡ10mol/L；⑥缬沙-6坦组(Valsartangroup)：预先加入缬沙坦10mol/L，再加入AngⅡ-610mol/L。采用抗α-actin免疫组化鉴定心肌细胞；Image-ProPlus专业图像处理软件测定细胞表面积；考马斯亮蓝法测定细胞总蛋白含量；采用

5、p-NPP酶法测定心肌细胞肌浆网钙泵（Sarcoplasmicreticulum2+Ca-ATPases,SERCA）活力；定磷法测定钙调神经磷酸酶（Calcineurin,CaN）活性及RT-PCR法半定量测定CaNmRNA表达。结果：⑴心肌细胞鉴定：心肌细胞经抗α-actin免疫细胞化学染色后，细胞质内可见棕黄色丝状物，证实为心肌细胞。⑵心肌细胞表面积：与control组比较，AngII组的细胞表面积显著增32加（761.75±95.09vs498.27±67.31um,P<0.01），其余各组细胞

6、表面积的增加无统计学差异（P>0.05）。与AngII组相比，Valsartan组、Quer组、Isor2组及TFH组的细胞表面积分别减小（552.07±93.48vs761.75±95.09um,2P<0.01）、(584.22±109.42vs761.75±95.09um,P<0.01）、(611.02±149.38vs22761.75±95.09um,P<0.01）、(572.05±104.37vs761.75±95.09um,P<0.01）;但这四组之间差异无统计学意义（P>0.05）。⑶心肌细

7、胞总蛋白含量：与control组比较，AngII组的细胞总蛋白含量明显增加（4.61±1.42vs2.63±0.79ng/well,P<0.01），其余各组细胞总蛋白含量的增加无统计学差异（P>0.05）。与AngII组相比，Valsartan组与TFH组的细胞总蛋白含量分别减小(3.04±0.86vs4.61±1.42ng/well,P<0.01)、(3.10±0.52vs4.61±1.42ng/well,P<0.01);Isor组与Quer组的细胞总蛋白含量分别减小(3.25±0.94vs4.61±

8、1.42ng/well,P<0.05)、(3.41±0.75vs4.61±1.42ng/well,P<0.05);但这四组之间差异无统计学意义（P>0.05）。⑷肌浆网钙泵（SERCA）活力：与control组比较，AngII组的SERCA活力明显下降（2.36±0.99vs4.11±1.19umol/gpro·min,P<0.01），其余各组SERCA活力下降无统计学差异（P>0.05）。与AngII组相比，Valsartan组、Isor