人脐血单个核细胞静脉移植联合复方丹参滴丸对急性心肌梗死局灶炎症反应的影响

Effect of Compound Danshen Dripping Pill Combined with Intravenous Transplantation of Human Umbilical Cord Blood Mononuclear Cells on Local Inflammatory Response in the Myocardium of Rabbits with Acute Myocardial Infarction

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DOI：10.7661/CJIM.2013.11.1532

中文关键词: 复方丹参滴丸人脐血单个核细胞心肌梗死炎症反应肿瘤坏死因子-α

英文关键词:Compound Danshen Dripping Pill human umbilical cord blood mononuclear cell myocardial infarction inflammation tumor necrosis factor α

基金项目:2011年湖南省中医药管理局科研资助课题项目（No2100045）

作者	单位	E-mail
邓柳霞,艾旗,袁春菊
余国龙	中南大学湘雅医院心内科（长沙410008）	yuguolong123@yahoo.com.cn

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全文下载次数: 2021

中文摘要:

目的观察复方丹参滴丸（Compound Danshen Dripping Pill，CDDP）对急性心肌梗死（acute myocardial infarction，AMI）心肌组织炎症反应的影响，观察人脐血单个核细胞（human umbilical cord blood mononuclear cells，HUCBMCs）静脉移植联合CDDP对家兔AMI心肌组织炎症反应、炎性因子肿瘤坏死因子-α（TNF-α）表达及心功能的影响，并探讨其联合治疗对AMI可能的保护机制。方法对40只健康家兔结扎冠状动脉左前降支，制备AMI模型。40只建模成功家兔随机分为4组，即对照组、CDDP组、单纯移植组及联合治疗组，每组10只。对照组：术后24 h生理盐水0.5 mL耳缘静脉注射，生理盐水5 mL/天灌胃；CDDP组：术后24 h经耳缘静脉注入生理盐水0.5 mL，CDDP 270 mg/d溶入生理盐水5 mL灌胃；单纯移植组：术后24 h经耳缘静脉注入含3×107个绿色荧光蛋白（green fluorescent protein，GFP）标记HUCBMCs生理盐水0.5 mL，生理盐水5 mL/天灌胃；联合治疗组：术后24 h经耳缘静脉注入含3×107 个GFP标记HUCBMCs的生理盐水0.5 mL，CDDP 270 mg/d溶入生理盐水5 mL灌胃。每组随机选取5只家兔分别于移植术后1、4周超声心动图检测家兔心功能指标，包括左室射血分数（left ventricular ejection fraction， LVEF）及左室短轴缩短率（left ventricular fractional shorting， LVFS）；HE染色光镜下观察心肌病理变化和白细胞计数；免疫组化检测心肌组织TNF-α蛋白表达量；荧光显微镜观测GFP阳性细胞。结果（1）与对照组比较，CDDP组、单纯移植组、联合治疗组术后1、4周心功能指标LVFS及LVEF改善（P<0.05），其中联合治疗组心功能指标改善优于单纯移植组、CDDP组（P<0.05），而后两组组间比较差异无统计学意义（P>0.05）；（2）与对照组比较，CDDP组、单纯移植组、联合治疗组术后1、4周心肌组织白细胞计数及TNF-α蛋白表达均显著减低（P<0.05），且联合治疗组心肌组织白细胞计数及TNF-α蛋白表达均显著低于单纯移植组、CDDP组（P<0.05），而后两组组间比较差异无统计学意义（P>0.05）；（3）联合治疗组、单纯移植组术后1、4周均可见GFP阳性细胞散在分布于梗死周边区域，且联合治疗组细胞计数多于单纯移植组（P<0.05）。结论本研究提示HUCBMCs静脉移植联合CDDP治疗AMI，可提高移植细胞在心肌组织内的存活率，并进一步改善心功能。其机制可能与联合治疗减轻心肌局部炎症反应及抑制促炎因子TNF-α蛋白的表达作用增强有关。

英文摘要:

ObjectiveTo investigate effect of Compound Danshen Dripping Pill （CDDP） on the inflammatory response of the myocardium of acute myocardial infarction （AMI） rabbits， to observe the therapeutic effect of CDDP combined intravenous transplantation of human umbilical cord blood mononuclear cells （HUCBMCs） on inflammatory response， pro-inflammatory cytokine tumor necrosis factor α （TNF-α）， and heart function in the myocardium of AMI rabbits， and to explore the possible protective mechanisms of the combined therapy. MethodsThe AMI model was successfully established by ligation of the left anterior coronary artery （LAD） in 40 healthy rabbits. Then they were randomly divided into four groups， i.e.， the control group， the CDDP group， the transplantation group， and the combined group， 10 in each group. Rabbits in the control group received intravenous injection of 0.5 mL normal saline via ear vein within 24 h after AMI and then intragastric infusion of normal saline at 5 mL per day. Rabbits in the CDDP group received intravenous injection of 0.5 mL normal saline via ear vein within 24 h after AMI and then intragastric infusion of solution obtained by solving 270 mg CDDP in 5 mL normal saline per day. Rabbits in the transplantation group received intravenous injection of 0.5 mL normal saline labeled with green fluorescent protein （GFP） containing 3 × 107 of HUCBMCs via ear vein within 24 h after AMI and then intragastric infusion of normal saline at 5 mL per day. Rabbits in the combined group received intravenous injection of 0.5 mL normal saline labeled with GFP containing 3 × 107 of HUCBMCs via ear vein within 24 h after AMI and then intragastric infusion of solution obtained by solving 270 mg CDDP in 5 mL normal saline per day. At week 1 and 4 after treatment， cardiac function indices such as left ventricular fractional shorting （LVFS） and left ventricular ejection fraction （LVEF） were performed by echocardiography； the number of transplanted cells in the myocardium was found by GFP positive cells counted with fluorescence microscopy. The white blood cells in the myocardium stained with HE were determined by light microscope. The expressions of TNF-α protein in the myocardium were detected by immunohistochemical assay. Results（1） Compared with the control group at week 1 and 4 after treatment， the LVEF and LVFS were significantly improved in the CDDP， transplantation， and combined groups （P<0.05）. The cardiac function was significantly improved in the combined group than in the CDDP group and the transplantation group （P<0.05）. But there was no statistical difference in the latter two groups. （2） Compared with the control group， the number of white blood cells and the expression of TNF-α protein decreased significantly in the CDDP， transplantation， and combined groups at week 1 and 4 respectively after treatment. The number of white blood cells and expressions of TNF-α protein were significantly lower in the combined group than in the CDDP group and the transplantation group （P<0.05）. But there was no statistical difference in the latter two groups. （3） GFP-positive cells were found to be distributed in the peri-myocardial infarction area in the transplantation group and the combined group at week 1 and 4 after transplantation. Besides， the number of the GFP positive cells was much more in the combined group than in the transplantation group （P<0.05）. ConclusionsThe findings indicated that the combination of CDDP with intravenous transplantation of HUCBMCs in the treatment of AMI rabbits could elevate the survival rate of transplanted cells， and further improve the heart function. The possible mechanisms might be related to attenuating local inflammation of myocardium， and inhibiting enhanced expressions of pro-inflammatory cytokine TNF-α protein.

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