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、15 mg/kg;注射时间分为3个水平,分别为7、14、21 d.对照组注射等量的生理盐水(1 ml).在连续注射7、14、21 d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15 mg/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10 mg/kg组和15 mg/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CsA 7、14、21 d,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=7.14,P<0.05);其中10mg/kg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10 mg/kg组[(15.95±1.00)、(12.74±1.31)、(14.01±0.81)U/mg Pr]和15 mg/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10 mg/kg组[(6.91±0.70)%、(11.10±2.05)%、(19.81±5.00)%]和15 mg/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5 mg/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.
目的 觀察環孢黴素A(CsA)對大鼠心肌組織的損傷作用,為CsA在心髒移植患者中長期安全用藥提供科學依據.方法 Wistar大鼠84隻,雌性,體質量(200±25)g.按析因設計將大鼠按體質量隨機分為12組,每組7隻,腹腔註射CsA.註射劑量分為4箇水平,分彆為0(對照)、5、10、15 mg/kg;註射時間分為3箇水平,分彆為7、14、21 d.對照組註射等量的生理鹽水(1 ml).在連續註射7、14、21 d時處死大鼠,取心肌組織,光鏡、電鏡下觀察大鼠心肌細胞形態結構變化:測定心肌組織丙二醛(MDA)和超氧化物歧化酶(SOD)活力;末耑標記(TUNEL)法檢查心肌細胞凋亡,計算細胞凋亡指數(AI).結果 光鏡下,與對照組比較,小劑量CsA(5mg/kg組)對心肌組織無明顯影響;10mg/kg組和15 mg/kg組心肌組織齣現不同程度的水腫、變性、壞死,部分心肌細胞空泡變,心肌橫紋消失:電鏡下10 mg/kg組和15 mg/kg組心肌細胞可見到線粒體損傷、覈濃縮、覈邊集,部分線粒體齣現竈狀空化,肌質內可見部分高度擴張的內質網.在腹腔註射CsA 7、14、21 d,註射時間與註射劑量對大鼠心肌組織MDA的影響有統計學意義(F=6.37、10.15,P均<0.05);時間和劑量存在交互作用(F=7.14,P<0.05);其中10mg/kg組[(2.29±0.18)、(3.10±0.45)、(2.57±0.37)nmol/mg Pr]和15mg/kg組[(3.09±0.63)、(3.32±0.52)、(3.34±0.29)nmol/mg Pr]均明顯高于對照組[(1.98±0.20)、(2.04±0.52)、(1.99±0.26)nmol/mg Pr,P均<0.05].註射時間與註射劑量對大鼠心肌組織SOD活力的影響有統計學意義(F=8.43、11.69,P均<0.05);時間和劑量存在交互作用(F=9.86,P<0.05);其中10 mg/kg組[(15.95±1.00)、(12.74±1.31)、(14.01±0.81)U/mg Pr]和15 mg/kg組[(13.04±1.01)、(14.68±0.81)、(14.01±0.63)U/mg Pr]均明顯高于對照組[(10.38±0.80)、(9.73±0.58)、(10.20±0.26)U/mg Pr,P均<0.05].光鏡下凋亡細胞覈呈棕黃或褐色.註射時間與註射劑量對4組大鼠心肌組織AI的影響有統計學意義(F=10.02、20.46,P均<0.05);時間和劑量存在交互作用(F=15.73,P<0.05).其中10 mg/kg組[(6.91±0.70)%、(11.10±2.05)%、(19.81±5.00)%]和15 mg/kg組[(11.02±2.02)%、(15.51±1.31)%、(33.40±6.60)%]均明顯高于對照組[(4.40±0.13)%、(4.60±1.20)%、(5.20±1.10)%]和5 mg/kg組[(4.60±0.10)%、(5.00±2.11)%、(5.43±1.11)%,P均<0.05].結論 小劑量CsA對心肌組織無明顯影響,但較大劑量可緻心肌細胞損傷,引起氧化應激,誘導心肌細胞凋亡;器官移植後,應密切註意和避免長時間、大劑量作為免疫抑製劑應用的CsA所引起的心肌損傷.
목적 관찰배포매소A(CsA)대대서심기조직적손상작용,위CsA재심장이식환자중장기안전용약제공과학의거.방법 Wistar대서84지,자성,체질량(200±25)g.안석인설계장대서안체질량수궤분위12조,매조7지,복강주사CsA.주사제량분위4개수평,분별위0(대조)、5、10、15 mg/kg;주사시간분위3개수평,분별위7、14、21 d.대조조주사등량적생리염수(1 ml).재련속주사7、14、21 d시처사대서,취심기조직,광경、전경하관찰대서심기세포형태결구변화:측정심기조직병이철(MDA)화초양화물기화매(SOD)활력;말단표기(TUNEL)법검사심기세포조망,계산세포조망지수(AI).결과 광경하,여대조조비교,소제량CsA(5mg/kg조)대심기조직무명현영향;10mg/kg조화15 mg/kg조심기조직출현불동정도적수종、변성、배사,부분심기세포공포변,심기횡문소실:전경하10 mg/kg조화15 mg/kg조심기세포가견도선립체손상、핵농축、핵변집,부분선립체출현조상공화,기질내가견부분고도확장적내질망.재복강주사CsA 7、14、21 d,주사시간여주사제량대대서심기조직MDA적영향유통계학의의(F=6.37、10.15,P균<0.05);시간화제량존재교호작용(F=7.14,P<0.05);기중10mg/kg조[(2.29±0.18)、(3.10±0.45)、(2.57±0.37)nmol/mg Pr]화15mg/kg조[(3.09±0.63)、(3.32±0.52)、(3.34±0.29)nmol/mg Pr]균명현고우대조조[(1.98±0.20)、(2.04±0.52)、(1.99±0.26)nmol/mg Pr,P균<0.05].주사시간여주사제량대대서심기조직SOD활력적영향유통계학의의(F=8.43、11.69,P균<0.05);시간화제량존재교호작용(F=9.86,P<0.05);기중10 mg/kg조[(15.95±1.00)、(12.74±1.31)、(14.01±0.81)U/mg Pr]화15 mg/kg조[(13.04±1.01)、(14.68±0.81)、(14.01±0.63)U/mg Pr]균명현고우대조조[(10.38±0.80)、(9.73±0.58)、(10.20±0.26)U/mg Pr,P균<0.05].광경하조망세포핵정종황혹갈색.주사시간여주사제량대4조대서심기조직AI적영향유통계학의의(F=10.02、20.46,P균<0.05);시간화제량존재교호작용(F=15.73,P<0.05).기중10 mg/kg조[(6.91±0.70)%、(11.10±2.05)%、(19.81±5.00)%]화15 mg/kg조[(11.02±2.02)%、(15.51±1.31)%、(33.40±6.60)%]균명현고우대조조[(4.40±0.13)%、(4.60±1.20)%、(5.20±1.10)%]화5 mg/kg조[(4.60±0.10)%、(5.00±2.11)%、(5.43±1.11)%,P균<0.05].결론 소제량CsA대심기조직무명현영향,단교대제량가치심기세포손상,인기양화응격,유도심기세포조망;기관이식후,응밀절주의화피면장시간、대제량작위면역억제제응용적CsA소인기적심기손상.
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(AI) 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)nmol/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 < 0.05], 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 dose 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)nmol/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], respectively. Apoptosis nucleus appeared huffy or brown under the light microscope. The effects of different time and dose on AI of rat myocardium 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 < 0.05] 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.
