摘要
目的观察环孢霉素A（CsA）对大鼠心肌组织的损伤作用，为CsA在心脏移植患者中长期安全用药提供科学依据。方法Wistar大鼠84只，雌性，体质量（200±25）g。按析因设计将大鼠按体质量随机分为12组，每组7只，腹腔注射CsA。注射剂量分为4个水平，分别为0（对照）、5、10、15mg／kg；注射时间分为3个水平，分别为7、14、21d。对照组注射等量的生理盐水（1ml）。在连续注射7、14、21d时处死大鼠，取心肌组织，光镜、电镜下观察大鼠心肌细胞形态结构变化；测定心肌组织丙二醛（MDA）和超氧化物歧化酶（SOD）活力；末端标记（TUNEL）法检查心肌细胞凋亡，计算细胞凋亡指数（AI）。结果光镜下，与对照组比较，小剂量CsA（5mg／kg组）对心肌组织无明显影响；10mg／kg组和15mg／kg组心肌组织出现不同程度的水肿、变性、坏死，部分心肌细胞空泡变，心肌横纹消失；电镜下10mg／kg组和15mg／kg组心肌细胞可见到线粒体损伤、核浓缩、核边集．部分线粒体出现灶状空化，肌质内可见部分高度扩张的内质网。在腹腔注射CsA7、14、21d，注射时间与注射剂量对大鼠心肌组织MDA的影响有统计学意义（F=6．37、10．15，P均〈0．05）；时间和剂量存在交互作用（F=7A4，P〈0．05）；其中10ms／ks组[（2．29±0．18）、（3．10±0．45）、（2．57±0．37）nmol／mg Pr]和15mg／kg组[（3．09±0．63）、（3．32±0．52）、（3．34±0．29）nmol／mg Pr]均明显高于对照组[（1.98±0．20）、（2．04±0．52）、（1．99±0．26）nmol／mg Pr，P均〈0．05]。注射时间与注射剂量对大鼠心肌组织SOD活力的影响有统计学意义（F=8．43、11．69，P均〈0．05）；时间和剂量存在交互作用（F=9．86，P〈0．05）；其中10mg／kg组[（15．95±1．00）、（12．74±1．31）、（14．01±0．81）U／mg Pr]和15mg／kg组[（13．04±1．01）、（14．68±0．81）、（14．01±0．63）U／mg Pr]均明显高于对照组[（10．38±0．80）、（9．73±0．58）、（10．20±0．26）U／mg Pr，P均〈0．05]。光镜下凋亡细胞核呈棕黄或褐色。注射时间与注射剂量对4组大鼠心肌组织AI的影响有统计学意义（F=10．02、20．46，P均〈0．05）；时间和剂量存在交互作用（F=15．73，P〈0．05）。其中10mg／kg组[（6．91±0．70）％、（11．10±2．05）％、（19．81±5．00）％]和15mg／kg组[（11．02±2．02）％、（15．51±1．31）％、（33．40±6．60）％]均明显高于对照组[（4．40±0．13）％、（4．60±1．20）％、（5．20±1．10）％]和5mg／kg组[（4．60±0．10）％、（5．00±2．11）％、（5．43±1．11）％，P均〈0．05]。结论小剂量CsA对心肌组织无明显影响，但较大剂量可致心肌细胞损伤，引起氧化应激，诱导心肌细胞凋亡：器官移植后．应密切注意和避免长时间、大剂量作为免疫抑制剂应用的CsA所引起的心肌损伤。
Objective To observe the impairment of different doses of cyclosporine A（CsA） to the rat myocardial tissue to offer scientific evidence for the long-term safe application of CsA in heart transplantation. Methods Eighty-four female Wistar rats, each weighing of （200 ± 25）g, were randomly divided into 12 groups. On days 7,14,21 after a constant peritoneal injection of CsA （0,5,10,15 mg/kg） and 1 ml physiological saline in control group, the rats were put to death, the rat myocardial tissue taken, to observe the pathologic and structural changes of the tissue cells under light microscope and electron microscope. The contents of rat myocardium tissue malondialdehyde （MDA） and superoxide dismutase （SOD） were measured; cardiomyocyte apoptosis was detected and accounted, apoptosis index（M） was measured with the method of TUNEL. Results Small dose of CsA（5 mg/kg）had no obvious effects on cardiac tissue, in CsA groups of 10 mg/kg and 15 mg/kg, under the light microscope, there appeared edema, degeneration and necrosis of myocardium, part of cardiac myocyte had different level cavity; under the electron microscope, there appeared mitochondria damage, nucleus shrinkage and chromatic margination, part of cardiac myocyte had focus cavity. There was dilated endoplasic reticulum in the sarcoplasm. The effects of different time and dose on MDA content of rat myocardium tissue had statistical significance （F = 6.37,10.15, both P 〈 0.05）. Interaction between time and dose existed statistical significance （F= 7.14, P〈 0.05）. The MDA contents of CsA group of 10 mg/kg and 15 mg/kg were [（2.29 ± 0.18）, （3.10 ± 0.45）, （2.57 ± 0.37）nmol/L] and [ （3.09 ± 0.63）, （3.32 ± 0.52）, （3.34 ± 0.29）nmo]/L] on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [ （ 1.98 ± 0,20）, （2.04 ± 0.52）, （1.99 ± 0.26） nmol/L, all P 〈 0.051, respectively. The effects of different time and dose on SOD activity of rat myocardium tissue had statistical significance（F = 8.43,11.69, both P 〈 0.05）. Interaction between time and close existed statistical significance（F = 9.86, P 〈 0.05）. The SOD activity of CsA groups of 10 mg/kg and 15 mg/kg were （ 15.95 ± 1.00）, （ 12.74 ± 1.31 ）, （ 14.01 ± 0.81 ）nmot/L and （ 13.04 ± 1.01 ）, （ 14.68 ± 0.81 ）, （ 14.01 ± 0.63）nmol/L on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [ （ 10.38 ± 0.80）, （9.73 ± 0.58 ）, （ 10.20 ± 0.26）nmol/L, all P 〈 0.05 1, respectively. Apoptosis nucleus appeared huffy or brown under the light microscope. The effects of different time and dose on AI of rat myoeardium tissue had statistical significance （F = 10.02,20.46, both P 〈 0.05）. Interaction between time and dose existed statistical significance （F = 15.73,P 〈 0.05）. The AI of CsA groups of 10 mg/kg and 15 mg/kg were （6.91 ± 0.70）%, （11.10± 2.05）%,（19.81 ± 5.00）% and （11.02 ± 2.02）%,（15.51 ± 1.31）%,（33.40 ± 6.60）% on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [（4.40 ± 0.13）%, （4.60 ± 1.20）%, （5.20 ± 1.10）, all P 〈 0.051 and CsA group of 5 mg/kg [（4.60 ± 0.10）%, （5.00 ± 2.11 ）%, （5.43 ± 1.11 ）%, all P 〈 0.05], respectively. Conclusion Small dose of CsA has no obvious effects on cardiac tissue, but large dosage can induce myocyte apoptosis and damage by causing oxidative stress; after implantation, attention should be paid to cardiac impairment due to constant large dosage of CsA.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2009年第4期413-416,共4页
Chinese Jouranl of Endemiology
基金
国家自然科学基金（30872387）
高等学校博士学科点专项科研基金（200802260006）
黑龙江省研究生创新基金（YJSCX2007-0204HLJ）
