粒细胞集落刺激因子与干细胞因子合用对急性心肌梗死模型大鼠心功能的影响

背景:骨髓干细胞动员以其无创伤、无免疫排斥反应,不需要干细胞提取、培养、扩增等工作倍受青睐。目的:观察联合应用粒细胞集落刺激因子和干细胞因子对大鼠急性心肌梗死的治疗作用。设计:随机对照观察。单位:锦州医学院药理教研室实验室。材料:选用32只成年雄性SD大鼠,体质量(200±20)g,购自锦州医学院实验动物中心。重组人粒细胞集落刺激因子(北京双鹭制药),重组人干细胞因子(成都地奥有限公司)。方法:实验于2005-07/11在锦州医学院药理教研室实验室进行。①5mg/kg盐酸异丙基肾上腺素左下腹腔注射复制急性心肌梗死模型,3h后将大鼠随机分为4组,每组8只。综合治疗组:用生理盐水稀释浓度至10mg/L的重组人粒细胞集落刺激因子1mL/kg和重组人干细胞因子1mL/kg皮下注射,连续5d。粒细胞集落刺激因子组:单用生理盐水稀释浓度至10mg/L的rhG-CSF1mL/kg皮下注射,每只大鼠和综合治疗组大鼠注射药品差量用生理盐水(1mL/kg)补足,连续5d。干细胞因子组:单用生理盐水稀释浓度至10mg/L的人干细胞因子1mL/kg皮下注射,每只大鼠和综合治疗组大鼠注射药品差量用生理盐水(1mL/kg)补足,连续5d。对照组:单用生理盐水(2mL/kg)皮下注射。②分别于造模后第14d和28d,摸球法从每组大鼠中抽取4只麻醉,自左心室心尖部插入内充肝素钠的塑料管至左心腔,用Medlab-u18c生物信号采集处理系统描记左心室峰压,左心室舒张末压,左室内压最大上升/下降速度,并同步记录心率。拔出气管插管,立即剪掉心脏,剪掉心房及右心室,微量天平称重,计算左心室质量/体质量,评估心肌重塑情况,将组织块切片,光镜下观测大鼠心肌病理形态及梗死面积。采用细胞图像分析仪病理图文分析每组大鼠心肌梗死面积。主要观察指标:各组大鼠左心功能指标、心肌梗死面积、心率和心室重塑情况及大鼠心肌病理形态。结果:纳入大鼠32只全部进入结果分析,无脱落。①大鼠左心功能指标比较结果:综合治疗组大鼠造模14,28d后左心室收缩压、左心室舒张末压、左心室内压最大上升/下降速度高于其他3组,造模28d时上述各指标均高于14d,差异有统计学意义(P<0.05),粒细胞集落刺激因子组两时间点上述指标均大于干细胞因子组及对照组(P<0.05)。②大鼠两时间点心肌梗死面积、心率和心室重塑情况:综合治疗组大鼠造模14,28d后心肌梗死面积及左心室重量小于其他3组,造模后28d心肌梗死面积小于14d(P<0.05),粒细胞集落刺激因子组两时间点上述指标均小于干细胞因子组与对照组。③大鼠心肌病理形态改变:造模后14及28d,综合治疗组和粒细胞集落刺激因子组均未见明显心肌瘢痕组织,有新生毛细血管生长,粒细胞集落刺激因子组毛细血管密度小于综合治疗组,均有成纤维细胞浸润。干细胞因子组和对照组可见瘢痕组织呈散在的灶性分布,有成堆或散在分布的淋巴细胞,其核小、深染、胞浆少,细胞间胞浆融合,新生肉芽组织增生不明显。结论:粒细胞集落刺激因子和干细胞因子合用对急性心肌梗死大鼠缺血损伤心肌的保护和再生作用优于单用,可改善急性心肌梗死大鼠的心室功能。

Effects of granulocyte colony-stimulating factor plus stem cell factor on the cardiac function of rats with acute myocardial infarction

BACKGROUND: Mobilizing bone stem cell is popular because of no hurling, no immunological rejection and no extraction,cultivation and amplification of stem cells.OBJECTIVE: To investigate the effects of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) on treating acute myocardial infarction (AMI) of rats.DESIGN: Randomized controlled study.SETTING: Laboratory of Pharmacology, Jinzhou Medical College.MATERIALS: A total of 32 healthy SD rats weighing (200+20) g were provided by the Animal Center of Jinzhou Medical College. Recombination human G-CSF (rhG-CSF) was provided by Shuanglu medicine factory, Beijing, and recombination human SCF (rhSCF) was provided by Diao Company, Chengdu.METHODS: The experiment was carried out in Jinzhou Medical Pharmacological Laboratory from July to November 2005.① Totally, 32 male SD rats were injected with 5 mg/kg ISO in left abdomen. After 3 hours, the rats were randomly divided into 4 groups with 8 in each group. Treatment group: The rats were injected with 1 mg/kg rhG-CSF and 1 mg/kg rhSCF under epidermis for 5 days. The concentration was 10 mg/kg. G-CSF group: The rats were injected with 1 mg/kg rhG-CSF and 1 mg/kg saline for 5 days. The concentration was 10 mg/kg. SCF group: The rats were injected with 1 mg/kg rhSCF and 1 mg/kg saline for 5 days. The concentration was 10 mg/kg. Control group: 2 mg/kg saline was injected under epidermis. ② After 14 days and 28 days from injecting ISO, one subgroup was weighted from every group.The rats were narcotized with 200 g/L urethane. After the heart's exposed, a tube filled with heparin sodium was inserted to left ventricle. Then tracked record the left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximal rising and descent velocity of left ventricular pressure (±dp/dr) using Medlab-u/Sc and kept the heart rate (HR) records. Tracheal cannula was pushed out; heart was cut off immediately; auricula and right ventricle were cut off; tissue was weighed with microbalance; left ventricle mass/body mass was calculated; myocardial remodeling was evaluated. Tissues were cut into sections, and pathological form and infarction area were observed under optic microscope. Infarction area in each group was analyzed with cellular image analyzer.MAIN OUTCOME MEASURES: Left ventricular function, myocardial infarction area, heart rate, ventricular remodeling and pathology of the cardiac muscle.RESULTS: All 32 rats were involved in the final analysis without any loss. ① Comparison of the left ventricular function:At 14 days after modeling, LVSP, LVEDP and ±dp/dr were higher in treatment group than those in other 3 groups (P ＜0.05); 28 days later, those indexes in the 4 groups were higher than those in the 4 groups at 14 days after modeling,and there was significant difference (P ＜ 0.05). Those indexes in G-CSF group were higher than those in SCF group and control group at the two time points (P ＜ 0.05). ② Comparison of infarction area, HR and the ventricular remodeling: At 14 and 28 days after modeling, infarction area and left ventricle weight were smaller and lighter in treatment group than those in other 3 groups (P ＜ 0.05); 28 days later, infarction area in the 4 groups was smaller than those in the 4 groups at 14 days after modeling (P ＜ 0.05). Those indexes in G-CSF group were less than those in SCF group and control group at the two time points. ③ Pathology results: At 14 and 28 days after modeling, there was no significant infarction car in treatment group and G-CSF group, and new capillary vessel and infiltration of fibroblast were found in the two groups. The density of capillary vessel in G-CSF group was less than that in treatment group. There were infarction scars and much lymphocyte in SCF group and control group. The nucleus was small, the color was deep, and plasma was less. New granulation tissue was not significant.CONCLUSION: Combining G-CSF with SCF can protect the ischemic cardial muscle and improve the heart function in rats. The function of combining G-CSF and SCF is better than singlly using G-CSF or SCF.