聚合人脐带血红蛋白对大鼠心肌组织影响的初步探讨
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摘要
目的探讨聚合人脐带血红蛋白(PolyCHb)对动物心肌组织的影响。方法取18只健康雄性SD大鼠随机均分为假手术(Sham)组:大鼠麻醉后仅行股动、静脉插管,颈静脉及右侧颈总动脉插管;NaCl组及PolyCHb组:除同Sham组一样麻醉、插管后,分别输注生理盐水或PolyCHb(800 mg/kg);连续监测基础值、输注后0、10、20、30、60 min 3组大鼠的平均动脉压(MAP)、心率(HR)、左心室收缩末期压力(pes)、左心室舒张末期压力(ped)、室内压最大上升速率(dp/dt max)、室内压最大下降速率(dp/dt min)、等容舒张时间常数(Tau)。另取18只大鼠同样分作3组,麻醉、插管后按分组做相应处理,取输后1、3、6 h血样于EDTA抗凝管中,离心取血浆,测定肌酸激酶同工酶(CK-MB)及肌钙蛋白Ⅰ(cTnⅠ)含量,6 h时处死大鼠,取心脏做病理学检测。结果 PolyCHb组、 NaCl组与sham组比较:1)术(输)后0—60 min MAP(mmHg)及pes(mmHg)持续升高,至输注后60 min时,分别为128.50±7.26 vs 115.67±4.72 vs 106.5±14.84(P<0.05)与148.30±18.16 vs 122.83±13.57 vs 122.07±19.00(P<0.05);术(输)后HR(次/min):0 min时分别为302.83±20.76vs 371.00±43.36 vs 387.50±47.27 (P<0.05), 60 min时分别为391.50±34.26 vs 416.83±33.47 vs 431.50±33.76 (P>0.05); ped(mmHg):60 min时分别为11.34±11.58 vs 6.83±4.79 vs 8.14±4.25(P>0.05);2)dp/dt max(mmHg/s)与dp/dt min(mmHg/s)术(输)注0 min分别为6 530.50±418.63 vs 8 341.50±1 095.00 vs 8 242.17±1 579.38与5 352.83±449.02 vs 7 687.00±1 111.87 vs 7 799.50±2 246.39(P<0.05),至术(输)后60 min分别为8 139.00±758.26 vs 10 083.33±1 256.59 vs 10 032.17±1 779.30与7 457.33±915.62 vs 9 192.00±892.90 vs 9 069.50±2 104.49(P<0.05);3)Tau术(输)后0 min分别为17.01±3.72 vs 12.69±3.32 vs 12.57±2.07(P<0.05);4)CK-MB(pg/mL)术(输)后1、3 h术(输)后1、3 h时分别为63.41±13.15 vs 49.67±20.10 vs 38.20±6.02与101.88±28.08 vs 37.80±18.21 vs 37.94±10.71(P<0.05);5)cTnI(pg/mL)术(输)后3 h分别为14.61±7.03 vs 5.32±2.96 vs 5.72±2.90(P<0.05)。PolyCHb组大鼠术(输)后6 h病理检查未见心肌损伤。结论输注PolyCHb可影响心肌功能,酶学水平升高,但不会造成病理损伤。
Objective To investigate the effects of polymerized human cord hemoglobin(PolyCHb) on myocardial tissue in animals.Methods Eighteen healthy male Sprague-Dawley rats were randomly divided into sham operation group,NaCl group and PolyCHb group. Rats in sham operation group were anesthetized with femoral artery, venous cannula, jugular vein and right common carotid artery cannula; rats in NaCl group and PolyCHb group were infused with saline or PolyCHb(800 mg/kg) separately besides anesthesia and intubation conducted in the sham group; the mean arterial pressure was continuously monitored at the baseline, 0, 10, 20, 30, and 60 min after infusion. Mean arterial pressure(MAP), heart rate(HR), left ventricular end-systolic pressure(pes), left ventricular end-diastolic pressure(ped), maximum rate of increase in indoor pressure(dp/dt max), maximum rate of decline in indoor pressure(dp/dt min), isovolumic relaxation time constant(Tau) of the rats in three groups were tested. Another 18 rats were also divided into 3 groups. After anesthesia and intubation, the rats were treated according to the group. The blood samples were taken in the EDTA anticoagulation tube at 1, 3, and 6 h after infusion, and the plasma was centrifugated to determine the creatine kinase isoenzyme(CK-MB) and troponin I(cTnI) levels.The rats were sacrificed at 6 h, and the heart was taken for pathological examination. Results The related parameters in PolyCHb group, the NaCl group and the sham group were compared as follows: 1) MAP(mmHg) and pes(mmHg) continued to increase during 0—60 min after surgery(or infusion), and reached to 128.50±7.26 vs. 115.67±4.72 vs. 106.5±14.84(P<0.05) and 148.30±18.16 vs. 122.83±13.57 vs.122.07±19.00(P<0.05), respectively, at 60 min; HR(number of times/min) at 0 and 60 min after surgery(or infusion) was 302.83±20.76 vs. 371.00 ±43.36 vs. 387.50±47.27(P<0.05) and 391.50±34.26 vs. 416.83±33.47 vs. 431.50±33.76(P>0.05); ped(mmHg) at 60 min was 11.34±11.58 vs. 6.83±.4.79 vs. 8.14±4.25(P>0.05); 2) dp/dt max(mmHg/s) and dp/dt min(mmHg/s) at 0 min were 6350.50 ±418.63 vs. 8341.50 ±1095.00 vs. 8242.17 ±1579.38 and 5352.83 ±449.02 vs. 7687.00 ±1111.87 vs. 7799.50 ±2246.39(P<0.05); at 60 min after surgery(or infusion) were 8 139.00 ±758.26 vs. 10 083.33 ±1256.59 vs. 10032.17 ±1779.30 and 7 457.33 ±915.62 vs 9192.00±892.90 vs. 9069.50±2104.49(P < 0.05); 3) Tau at 0 min was 17.01±3.72 vs. 12.69±3.32 vs. 12.57±2.07(P <0.05); 4) CK-MB(pg/mL) at 1 and 3 h after surgery(or infusion) was 63.41±13.15 vs. 49.67±20.10 vs. 38.20±6.02 and 101.88±28.08 vs. 37.80±18.21 vs. 37.94±10.71,respectively(P<0.05); 5) cTnI(pg/mL) at 3 h after surgery(or infusion) was 14.61 ±7.03 vs. 5.32 ±2.96 vs. 5.72 ±2.90(P<0.05). No myocardial injury at 6 h after surgery(or infusion) was observed in the PolyCHb group.Conclusion Infusion of PolyCHb can affect myocardial function and increase the enzymology level, but it will not cause pathological damage.
Objective To investigate the effects of polymerized human cord hemoglobin(PolyCHb) on myocardial tissue in animals.Methods Eighteen healthy male Sprague-Dawley rats were randomly divided into sham operation group,NaCl group and PolyCHb group. Rats in sham operation group were anesthetized with femoral artery, venous cannula, jugular vein and right common carotid artery cannula; rats in NaCl group and PolyCHb group were infused with saline or PolyCHb(800 mg/kg) separately besides anesthesia and intubation conducted in the sham group; the mean arterial pressure was continuously monitored at the baseline, 0, 10, 20, 30, and 60 min after infusion. Mean arterial pressure(MAP), heart rate(HR), left ventricular end-systolic pressure(pes), left ventricular end-diastolic pressure(ped), maximum rate of increase in indoor pressure(dp/dt max), maximum rate of decline in indoor pressure(dp/dt min), isovolumic relaxation time constant(Tau) of the rats in three groups were tested. Another 18 rats were also divided into 3 groups. After anesthesia and intubation, the rats were treated according to the group. The blood samples were taken in the EDTA anticoagulation tube at 1, 3, and 6 h after infusion, and the plasma was centrifugated to determine the creatine kinase isoenzyme(CK-MB) and troponin I(cTnI) levels.The rats were sacrificed at 6 h, and the heart was taken for pathological examination. Results The related parameters in PolyCHb group, the NaCl group and the sham group were compared as follows: 1) MAP(mmHg) and pes(mmHg) continued to increase during 0—60 min after surgery(or infusion), and reached to 128.50±7.26 vs. 115.67±4.72 vs. 106.5±14.84(P<0.05) and 148.30±18.16 vs. 122.83±13.57 vs.122.07±19.00(P<0.05), respectively, at 60 min; HR(number of times/min) at 0 and 60 min after surgery(or infusion) was 302.83±20.76 vs. 371.00 ±43.36 vs. 387.50±47.27(P<0.05) and 391.50±34.26 vs. 416.83±33.47 vs. 431.50±33.76(P>0.05); ped(mmHg) at 60 min was 11.34±11.58 vs. 6.83±.4.79 vs. 8.14±4.25(P>0.05); 2) dp/dt max(mmHg/s) and dp/dt min(mmHg/s) at 0 min were 6350.50 ±418.63 vs. 8341.50 ±1095.00 vs. 8242.17 ±1579.38 and 5352.83 ±449.02 vs. 7687.00 ±1111.87 vs. 7799.50 ±2246.39(P<0.05); at 60 min after surgery(or infusion) were 8 139.00 ±758.26 vs. 10 083.33 ±1256.59 vs. 10032.17 ±1779.30 and 7 457.33 ±915.62 vs 9192.00±892.90 vs. 9069.50±2104.49(P < 0.05); 3) Tau at 0 min was 17.01±3.72 vs. 12.69±3.32 vs. 12.57±2.07(P <0.05); 4) CK-MB(pg/mL) at 1 and 3 h after surgery(or infusion) was 63.41±13.15 vs. 49.67±20.10 vs. 38.20±6.02 and 101.88±28.08 vs. 37.80±18.21 vs. 37.94±10.71,respectively(P<0.05); 5) cTnI(pg/mL) at 3 h after surgery(or infusion) was 14.61 ±7.03 vs. 5.32 ±2.96 vs. 5.72 ±2.90(P<0.05). No myocardial injury at 6 h after surgery(or infusion) was observed in the PolyCHb group.Conclusion Infusion of PolyCHb can affect myocardial function and increase the enzymology level, but it will not cause pathological damage.
引文
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[9] 闫东,李遥金,郝莎莎,等.应用大鼠血液置换模型对新型携氧代血浆功效的初步实验观察.中国输血杂志,2015,28(3):260-263.
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[11] 桑培培,李遥金,段炼,等.羟乙基淀粉加多聚人胎盘血红蛋白对失血性休克大鼠肠缺血再灌注损伤的影响,中国输血杂志,2017,30(4):358-362.
[12] Zhou WT,Li S,Liu JX,et al.An optimal polymerization process for low mean molecular weight HBOC with lower dimer.Artif Cells Blood Substit Immobil Biotechnol,2015,43(3):148-151.
[13] Pacher P,Nagayama T,Mukhopadhyay P,et al.Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.Nature Protocols,2008,3(9):1422-1434.
[14] Sagawa K.The ventricular pressure-volume diagram revisited.Circ Res,1978,43(5):677-687.
[15] Aoieong ESY,Williams A,Jani V,Cabrales P,et al.Cardiac function during resuscitation from hemorrhagic shock with polymerized bovine hemoglobin-based oxygen therapeutic.Artificial Cells,Nanomed Biotechnol,2017,45(4):686-693.
[16] Yu B,Bloch KD,Zapol WM.Hemoglobin-based red blood cell substitutes and nitric oxide.Trends Cardiovas Med,2009,19(3):103-107.
[17] 李涛,杨成民,刘进.血红蛋白类携氧载体心脏作用的两面性及其机理.中国输血杂志.2012,25(1):81-84.
[18] 文亚莉.心肌酶谱与血清肌钙蛋白Ⅰ联合检测在诊断急性心肌梗死中的作用.山东医药,2011,51(49):111-112.
[19] 马自梅.心肌肌钙蛋白Ⅰ在急性心肌损伤诊断中的临床应用价值.齐齐哈尔医学院学报,2012,33(6):759-760.
[20] Kenneth B,Donovan G,Timothy E.Review of hemoglobin-induced myocardial lesions.Artif cells Blood Substit Biotechnol,2004,32(3):353-374.
[21] Chi MH.Systems Biology of HBOC-induced vasoconstriction.Curr Drug Discov Technol,2012,9(3):204-211.
[2] Alayash AI.Blood substitutes:why haven′t we been more successful?Trends Biotechnol,2014,32(4):177-185.
[3] Vandegriff KD,Malavalli A,Minn C,et al.Oxidation and haem loss kinetics of poly(ethylene glycol)-conjugated haemoglobin (MP4):dissociation between in vitro and in vivo oxidation rates.Biochem J,2006,399(3):463-471.
[4] Wang Q,Sun L,Ji S,et al.Reversible protection of Cys-93(β) by PEG alters the structural and functional properties of the PEGylated hemoglobin.Biochim Biophys Acta (BBA)—Proteins and Proteomics,2014,1844(7):1201-1207.
[5] Yu B,Raher MJ,Volpato GP,et al.Inhaled nitric oxide enables artificial blood transfusion without hypertension .Circulation,2008,117(15):1982-1990
[6] Katz LM,Manning JE,McCurdy S,et al.Nitroglycerin attenuates vasoconstriction of HBOC-201 during hemorrhagic shock resuscitation.Resuscitation,2010.81(4):481-487.
[7] Li T,Yang G,Zhu Y,et al.Beneficial effects of novel cross-linked hemoglobin YQ23 on hemorrhagic shock in rats and pigs J Surg Res,2017;210:213-22.
[8] Li YJ,Yan D,Hao SS,et al.Polymerized human placenta hemoglobin improves resuscitative efficacy of hydroxyethyl starch in a rat model of hemorrhagic shock.Artif Cells Nanomed Biotechnol,2015,43(3):174-179.
[9] 闫东,李遥金,郝莎莎,等.应用大鼠血液置换模型对新型携氧代血浆功效的初步实验观察.中国输血杂志,2015,28(3):260-263.
[10] 蒋冰玉,李遥金,郝莎莎,等.携氧代血浆扩容及携氧功能初步动物实验研究.中国输血杂志,2016,29(4):357-360.
[11] 桑培培,李遥金,段炼,等.羟乙基淀粉加多聚人胎盘血红蛋白对失血性休克大鼠肠缺血再灌注损伤的影响,中国输血杂志,2017,30(4):358-362.
[12] Zhou WT,Li S,Liu JX,et al.An optimal polymerization process for low mean molecular weight HBOC with lower dimer.Artif Cells Blood Substit Immobil Biotechnol,2015,43(3):148-151.
[13] Pacher P,Nagayama T,Mukhopadhyay P,et al.Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.Nature Protocols,2008,3(9):1422-1434.
[14] Sagawa K.The ventricular pressure-volume diagram revisited.Circ Res,1978,43(5):677-687.
[15] Aoieong ESY,Williams A,Jani V,Cabrales P,et al.Cardiac function during resuscitation from hemorrhagic shock with polymerized bovine hemoglobin-based oxygen therapeutic.Artificial Cells,Nanomed Biotechnol,2017,45(4):686-693.
[16] Yu B,Bloch KD,Zapol WM.Hemoglobin-based red blood cell substitutes and nitric oxide.Trends Cardiovas Med,2009,19(3):103-107.
[17] 李涛,杨成民,刘进.血红蛋白类携氧载体心脏作用的两面性及其机理.中国输血杂志.2012,25(1):81-84.
[18] 文亚莉.心肌酶谱与血清肌钙蛋白Ⅰ联合检测在诊断急性心肌梗死中的作用.山东医药,2011,51(49):111-112.
[19] 马自梅.心肌肌钙蛋白Ⅰ在急性心肌损伤诊断中的临床应用价值.齐齐哈尔医学院学报,2012,33(6):759-760.
[20] Kenneth B,Donovan G,Timothy E.Review of hemoglobin-induced myocardial lesions.Artif cells Blood Substit Biotechnol,2004,32(3):353-374.
[21] Chi MH.Systems Biology of HBOC-induced vasoconstriction.Curr Drug Discov Technol,2012,9(3):204-211.