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Original Experimental Research
Journal of Chinese Integrative Medicine: Volume 2   July, 2004   Number 4

DOI: 10.3736/jcim20040414
Effect of soybean isoflavones on heart function of rats with adriamycin-induced heart failure
1. MA Shan-Feng (Department of Physiology, Bengbu Medical College, Bengbu, Anhui Province 233003, China E-mail: msf723@163.com)
2. GUAN Su-Dong (Department of Physiology, Bengbu Medical College, Bengbu, Anhui Province 233003, China )
3. ZHU Yan (Department of Physiology, Bengbu Medical College, Bengbu, Anhui Province 233003, China )

Objective: To observe the protective effect of soybean isoflavones (SI) on the heart function of the rats with adriamycin-induced heart failure.

Methods: Thirty adult male SD rats were divided into 5 groups: normal control (NC) group, adriamycin (ADR) group, L-SI group, M-SI group and H-SI group. SI of 30, 60, 120 mg·kg-1·d-1was orally administered through a stomach tube once a day for 6 days in L-SI group, M-SI group and H-SI group, respectively. The other two groups were given the same amount of normal saline the same way. Then ADR of 10 mg/kg was given intraperitoneally once to copy the model of heart-failure. The MedLab-U/4c biological signal collecting system was used to record and analyze the LVSP of the rats. The pathological changes of the cardiomyocytes were observed.

Results: As compared with NC group, the LVSP, ±dp/dtmax, Vpm of the ADR group were significantly lower (P<0.05 or P<0.01), but those of the H-SI group were markedly higher than those of the ADR group (P<0.01). Electron microscopic morphometry of the heart samples of the rats in ADR group revealed typical alterations, consisting an increase of collagen content, vacuolation, diminishing of the cardiomyocyte diameter, alteration of myofilaments and Z-lines of myofibers, and myofibrillar degeneration. SI of 120 mg·kg-1·d-1treatment could prevent the loss of myofibrillae and the reduction of myocyte diameter, and the degeneration of myofilaments and Z-lines were reversed by SI.

Conclusion: SI of 120 mg·kg-1·d-1 treatment can relieve the toxic effect of ADR on myocardium, and also obviously improve the cardiac contractility of heart-failure rats.

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J Chin Integr Med, 2004, 2(4): 278-280

作者简介: 马善峰(1971-),男,讲师.

Correspondence to: MA Shan-Feng, Lecturer. E-mail: msf723@163.com 

基金项目: 安徽省教育厅自然科学研究计划项目(2003KJ003zc)

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      阿霉素(adriamycin, ADR)是广谱抗肿瘤药物,被广泛用于治疗实质性及血液系统恶性肿瘤,但因明显的心肌毒性而使其临床应用受到限制。目前认为,雌激素对心血管具有保护作用[1]。植物雌激素是具有雌激素效能的天然化合物,而无人工雌激素的副作用,因而其预防和治疗心血管系统疾病的作用越来越受人们的重视[2,3]。大豆异黄酮(soybean isoflavones, SI)是植物雌激素的一种,本文旨在探讨其对阿霉素心衰大鼠心功能的影响。

 
  

1   材料与方法
1.1   材料
   (1)动物:健康雄性SD大鼠30只,体重190 ~210 g,由本院实验动物科提供。(2)大豆异黄酮,大连绿峰天然生物制品有限公司产品(批号:20021010);盐酸阿霉素,深圳万乐药业有限公司生产(批号:021101)。(3)主要仪器:MedLab-U/4c生物信号采集处理系统,南京美易科技有限公司生产。
1.2   造模与给药方法   大鼠随机分为5组,即空白对照组(NC),阿霉素组(ADR),大豆异黄酮低、中、高剂量组(L-SI、M-SI、H-SI),每组6只。具体处理如下:大豆异黄酮低、中、高剂量组分别予0.3%、0.6%、1.2%的大豆异黄酮10 ml ·kg-1·d-1灌胃;其余组每日同时灌胃等量的生理盐水,连续6 d。第7天,除对照组外,其余4组分别一次性腹腔注射盐酸阿霉素10 mg/kg,复制急性心衰模型[4]
1.3   实验方法   造模24 h后,用20%乌拉坦5 ml/kg腹腔注射麻醉。手术剥离气管并插管,同时游离出右侧颈总动脉,经其插入自制的心室插管(直径1 mm,充满1%肝素),描记血压曲线;再继续插入,使其通过左侧动脉瓣进入左心室,描记室内压曲线,自动分析处理左室收缩压(LVSP),心室内压最大上升速率(+dp/dtmax),心室内压最大下降速率(-dp/dtmax)和实测心肌最大收缩速度(Vpm)等数据。
1.4   标本制备   将动物处死,取心肌切成1 mm3小块,2.5%戊二醛固定,电镜观察超微结构变化。
1.5   统计学方法   全部数据均采用`x±s 表示,SPSS 11.5统计软件进行方差分析,最小有效差别(LSD)检验进行每两组之间的比较。

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2   结 果
2.1   大鼠心功能变化
   与NC组相比,ADR组LVSP、±dp/dtmax 、Vpm显著降低(P<0.01),表明阿霉素造模成功,造模组大鼠发生显著心衰。与ADR组相比,大豆异黄酮各组LVSP、±dp/dtmax 、Vpm均有升高,H-SI组LVSP、±dp/dtmax和Vpm显著升高(P<0.05 或P<0.01),表明高剂量大豆异黄酮对阿霉素致心衰有较好的防治作用,可明显改善心衰大鼠的心脏功能。见表1。

表1   各组大鼠左心功能指标
Tab 1   Effect of soybean isoflavones and adriamycin on heart function of rats

                                                                                                                  (`x±s )
Group
n
LVSP
(mm Hg)
+dp/dtmax
(mm Hg/s)
-dp/dtmax
(mm Hg/s)
Vpm
(/s)
NC
6
82.9±15.1
8 054.3±616.5
-4 754.8±752.5
1 676.4±243.2
ADR
6
54.9±12.5* *
3 443.8±654.3**
-2 494.9±674.7**
722.0±287.7**
L-SI
6
56.6±10.7**
3 555.5±731.0**
-2 563.7±359.2**
811.5±163.2**
M-SI
6
69.7±19.3
4 013.1±864.3**
-3 282.2±538.9**
1 027.0±282.3**
H-SI
6
74.7±14.1
4 829.8±1 289.1**
-3 649.9±821.9**△△
1 233.5±311.4**△△
**P< 0.01 vs NC group; P<0.05,△△ P< 0.01 vs ADR group

2.2   各组大鼠心肌超微结构变化   NC组:肌小节Z线排列整齐;线粒体形状规则,嵴丰富,排列规则。ADR组:心肌细胞内出现肌原纤维片状坏死、断裂、溶解,肌浆网扩张,肌小节失去正常结构;线粒体水肿,嵴减少或消失,甚至空泡变;间质中毛细血管内皮细胞突向管腔,胞浆中吞饮小泡增多,并可见较多的脂褐素沉积。L-SI组:心肌肌原纤维走行紊乱,出现不同方向的弯曲,肌小节长短不一,肌浆网囊状扩张;线粒体肿胀变性和局限性增多,有的嵴断裂和溶解,大量糖原颗粒沉积,基质疏松,电子密度降低呈空网状;毛细血管基底膜明显增厚;闰盘间隙明显增宽。M-SI组:肌原纤维灶性溶解,断裂,肌浆网扩张,线粒体数目增多,大小不一,部分线粒体水肿,嵴减少或消失,甚至空泡变。H-SI组:心肌病变明显减轻,仅部分肌原纤维排列紊乱,无溶解、坏死等病变,部分线粒体水肿。见图1。



图1   各组大鼠心肌超微结构(×10 K)
Fig 1   Ultrastructure of cardiomyocytes of the rat in different group (×10 K)

A: NC group; B: ADR group; C: L-SI group; D: M-SI group; E: H-SI group

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3   讨 论
     ADR致心衰主要是由于自由基的生成而引起膜脂质过氧化,心肌组织氧自由基的生成及脂质过氧化反应对心肌细胞膜完整性具有损害作用[5]。ADR可诱导细胞内外多种生化成分过氧化使心肌细胞内丙二醛(malondialdehyde, MDA)生成增多,超氧化物岐化酶(superoxide dismutase, SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-PX)和一氧化氮(nitric oxide, NO)活力下降。MDA可损伤生物膜,使膜酶受损,活性降低,产生心肌细胞变性坏死;SOD能有效清除氧自由基;GSH-PX不仅能清除羟自由基,还能降解脂质过氧化物[4];NO具有减轻心脏胶原蓄积,舒张血管和增加冠脉血流量的作用[6],NO可以作为氧自由基的内源性清除剂,减轻自由基的损伤,降低心肌细胞培养液的MDA含量[7],维持心肌收缩性,改善心肌缺血[8]。研究表明,雌激素使老年高血压动脉硬化患者血浆中的NO含量升高,对血管内皮有保护作用[9]
     本实验结果表明,SI能明显升高阿霉素心衰大鼠的LVSP、±dp/dtmax和Vpm,对阿霉素心衰有较好的治疗作用。电镜观察结果显示,阿霉素组大鼠心肌细胞线粒体水肿、破裂、嵴消失,甚至空泡变,表明有能量代谢障碍。脂褐素的沉积,说明有脂肪代谢异常。肌原纤维变性、溶解、断裂、大片坏死,说明心肌有肌溶性坏死。闰盘间隙增宽,心肌间质纤维化使心肌收缩、舒张功能严重受限。
     植物雌激素SI在体内代谢的主要活性物质为三羟异黄酮,能减弱细胞的过氧化状态,对心肌细胞有保护作用[10]。SI能使ADR引起的小鼠过氧化损伤心肌SOD、GSH-PX活性提高,过氧化脂质水平下降,有较强的抗氧化作用[11]。本实验提示SI能明显提高阿霉素心衰大鼠的LVSP、+dp/dtmax、-dp/dtmax和Vpm。经SI预防性治疗,心肌肌原纤维病变明显减轻,仅有部分肌原纤维排列紊乱,无肌原纤维坏死和溶解;仅部分线粒体水肿。因而SI对阿霉素致心衰有保护作用,在临床应用ADR进行肿瘤化疗时,SI可以作为一种减轻ADR心脏损伤的有效手段。但该作用是否通过 NO途径,以及有无其他途径尚待进一步的深入研究。

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References
1. Kaplan RC, Heckbert SR, Weiss NS, et al. Postmenopausal estrogens and risk of myocardial infarction in diabetic women[J]. Diabetes Care, 1998, 21(7) : 1117-1121.
    
2. Deodato B, Altavilla D, Squadrito G, et al. Cardioprotection by the phytoestrogen genistein in experimental myocardial ischemia-reperfusion injury[J]. Br J Pharmacol, 1999, 128(8) : 1683-1690.
    
3. Du Y, Xu Y. Medical value of isoflavones[J] Zhong Xi Yi Jie He Xue Bao, 2003, 1(4) : 296-300.
  
4. 金凯, 陈中军, 罗华荣. 盐酸阿霉素对大鼠心肌损伤的分子机制研究[J]. 同济大学学报(医学版), 2002, 23(3) : 191-194.
  
5. 钟玲, 叶秋荣. 刺五加对大鼠实验性缺血心肌的保护作用[J]. 中国病理生理杂志, 1995, 11(4) : 366-369.
  
6. Khaidar A, Marx M, Lubec B, et al. L-arginine reduces heart collagen accumulation in the diabetic db/db mouse[J]. Circulation, 1994, 90(1) : 479-483.
  
7. 万梅, 于占久. 内皮衍生舒张因子对缺血(缺氧)、再灌注(复氧)心肌的保护作用[J]. 生理学报, 1995, 47(3) : 231-237.
  
8. 董德武, 李丽颖, 魏宝霞. 葛根素与雌激素替代疗法对老年高血压、动脉粥样硬化患者血管内皮功能影响[J]. 中医药学报, 2002, 30(1) : 42-43.
  
9. 姜桢, 郭克芳, 董苏斐. 左旋精氨酸与氧自由基的相互作用[J]. 临床麻醉杂志, 2000, 16(10) : 503-505.
  
10. 闫祥华, 顾景范, 孙存普. 大豆异黄酮对大鼠血脂和过氧化状态的影响[J]. 营养学报, 2000, 22(1) : 31-35.
  
11. 郑高利, 朱寿民, 刘子贻, 等. 大豆异黄酮的抗氧化作用[J]. 浙江医科大学学报, 1997, 26(5) : 196-199.
  
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