心肌样细胞移植对大鼠心肌梗死的疗效观察

doi:10.3877/cma.j.issn.2095-1221.2017.04.003

中华细胞与干细胞杂志(电子版) ›› 2017, Vol. 07 ›› Issue (04) : 202 -207. doi: 10.3877/cma.j.issn.2095-1221.2017.04.003

所属专题：文献；

论著

心肌样细胞移植对大鼠心肌梗死的疗效观察

王秀力¹, 姜世强², 刘颖¹, 王洁¹, 吕茜¹, 王雷¹, 李春梅¹, 宫海滨¹^,^†(

)

^1. 221009　徐州市心血管病研究所　徐州市中心医院中心实验室　徐州市医学重点实验室
^2. 221009　徐州仁慈医院麻醉科

收稿日期:2016-12-19 出版日期:2017-08-01
通信作者: 宫海滨
基金资助:
江苏省生命健康科技专项资金资助项目(BL2012019); 徐州市科技发展项目(XM13B052)

Curative effect of transplantion of cardiomyocyte-like cells on myocardial infarction rats

Xiuli Wang¹, Shiqiang Jiang², Ying Liu¹, Jie Wang¹, Qian Lyu¹, Lie Wang¹, Chunmei Li¹, Haibin Gong¹^,^†()

^1. Department of Xuzhou Cardiovascular Disease Institute, Xuzhou Central Hospital Central Laboratory, Xuzhou Key Lab of Medica, Xuzhou 221009, China
^2. Department of Anesthesiology, Xuzhou Mercy Hospital, Xuzhou 221009, China

Received:2016-12-19 Published:2017-08-01
Corresponding author: Haibin Gong
About author:
Corresponding author: Gong Haibin, Email: ghbxzh@yahoo.com.cn

全文 ( ) 下载PDF ( ) 导出引用

引用本文：

王秀力, 姜世强, 刘颖, 王洁, 吕茜, 王雷, 李春梅, 宫海滨. 心肌样细胞移植对大鼠心肌梗死的疗效观察[J/OL]. 中华细胞与干细胞杂志(电子版), 2017, 07(04): 202-207.

Xiuli Wang, Shiqiang Jiang, Ying Liu, Jie Wang, Qian Lyu, Lie Wang, Chunmei Li, Haibin Gong. Curative effect of transplantion of cardiomyocyte-like cells on myocardial infarction rats[J/OL]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2017, 07(04): 202-207.

目的

观察心肌样细胞（CLC）移植对心肌梗死大鼠的疗效。

方法

制备大鼠心肌梗死动物模型，4周后超声检测大鼠血流动力学变化；体外分离、纯化大鼠骨髓间充质干细胞（BMSC），15 ng/ml的胰岛素样生长因子1（IGF-1）诱导BMSC分化为CLC 3，7，14和21 d，采用Western blot技术检测心肌标志性蛋白α-SMA，Ttroponin-T和Troponin-I的表达。CLC移植治疗心肌梗死大鼠，实验共分4组：心肌梗死组，生理盐水对照组，BMSC组及CLC组。移植治疗4周，超声检测血流动力学变化后取材大鼠心肌，冰冻切片，HE染色后检测大鼠心肌梗死的面积。统计分析采用单因素方差分析，实验组之间比较采用t检验。

结果

心肌梗死模型制备4周，模型组大鼠的LVSP数值为（118.74±11.56）mmHg，低于假手术组（130.42±12.58）mmHg，模型组大鼠的LVEDP数值为（14.29?±?7.63）mmHg，高于假手术组（2.44±1.11）mmHg，+dp/dt数值为（2645.17±468.16）mmHg/s低于假手术组（4087.29?±525.17）mmHg/s，-dp/dt数值为（2487.80±348.21）mmHg/s低于假手术组（3865.81±614.37）mmHg/s，差异均具有统计学意义（P < 0.01）。IGF-1诱导BMSC 3，7，14和21d后均检测到α-SMA、Troponin-T和Troponin-I的表达，与第3天比较P < 0.01。移植治疗4周后发现生理盐水对照组大鼠血流动力学没有任何改变，BMSC组及CLC组大鼠的LVEDP数值分别为（9.15±4.54）mmHg和（4.67±3.46）mmHg，低于心肌梗死组（14.29±7.63）mmHg，+dp/dt数值分别为（3245.45±368.76）mmHg/s和（3612.78±468.54）mmHg/s，高于心肌梗死组（2645.17±468.16）mmHg/s，-dp/dt数值分别为（3014.11±412.12）mmHg/s和（3347.21±398.56）mmHg/s，高于假手术组（2487.80±348.21）mmHg/s,同时CLC组大鼠的LVEDP（4.67±3.46）mmHg低于BMSC组（9.15±4.54）mmHg，±dp/dt数值分别是（3612.78±468.54）mmHg/s和（3347.21±398.56）mmHg/s高于BMSC组（3245.45±368.76）mmHg/s和（3014.11±412.12）mmHg/s，差异均具有统计学意义（P < 0.01）。BMSC组和CLC组的心梗面积与心肌梗死组比较明显减小，CLC组的梗死面积与BMSC组比较也明显减小，差异均具有统计学意义（P < 0.01）。

结论

IGF-1诱导BMSC分化为CLC后移植治疗心肌梗死大鼠比单独移植BMSC疗效更好。

关键词: 骨髓, 间质干细胞, 胰岛素样生长因子Ⅰ, 肌细胞，心脏, 心肌梗死, 细胞移植

Objective

To observe the effect of transplanting cardiomyocyte-like cells (CLC) to treat myocardial infarction in rats.

Methods

Animal models of myocardial infarction were prepared, and hemodynamics changes of rats models were tested by ultrasonic method 4 weeks later. Bone mesenchymal stem cells (BMSC) were isolated and purified in vitro, which were induced to CLCs by 15 ng/ml insulin-like growth factor 1 (IGF-1) , and then the expression of myocardial iconic protein α-SMA, Troponin-T and Troponin-Ⅰwere detected with Western blotting method in 3 d, 7 d, 14 d and 21 d. The experimental rats were randomly assigned into 4 groups (a myocardial infarction group, a normal saline group, a BMSC group and a CLC group) . The area of myocardial infarction was detected on rats 4 weeks after transplantation therapy by HE coloring. Data among multiple groups were compared by one-way ANOVA and data between two independent groups were compared by t-test.

Results

After myocardial infarction models were prepared for 4 weeks, the LVSP (118.74 ± 11.56) mmHg of the model group were significantly lower than those of the sham operation group (130.42 ± 12.58) mmHg, the LVEDP (14.29 ± 7.63) mmHg of the model group were significantly higher than those of the sham operation group (2.44 ±1.11) mmHg, +dp/dt (2645.17 ± 468.16) mmHg/s of the model group were significantly lower than those of the sham group (4087.29 ± 525.17) mmHg/s, and -dp/dt (2487.80 ± 348.21) mmHg/s of the model group were also significantly lower than those of the sham group (3865.81 ± 614.37) mmHg/s (P < 0.01) . The expressions of α-SMA, Troponin-T and Troponin-Ⅰwere detected when BMSCs were induced by IGF-1 on 3 d, 7 d, 14 d and 21 d (P < 0.01) . There were not any changes in haemodynamics of the normal saline group 4 weeks after transplantation therapy, the LVEDP of the BMSC group (9.15?±?4.54) mmHg and CLC group (4.67 ± 3.46) mmHg were significantly lower than those of the myocardial infarction group (14.29 ± 7.63) mmHg, the +dp/dt of BMSC group (3245.45?±?368.76) mmHg/s and CLC group (3612.78?±?468.54) mmHg/s were significantly higher than those of the myocardial infarction group (2645.17?±?468.16) mmHg/s, the -dp/dt of BMSC group (3014.11?±?412.12) mmHg/s and CLC group (3347.21?±?398.56) mmHg/s were also significantly higher than the sham group (2487.80?±?348.21) mmHg/s. In the meantime, the LVEDP of the CLC group (4.67?±?3.46) mmHg was significantly lower than that of the BMSC group (9.15?±?4.54) mmHg P < 0.01, the +dp/dt of the CLC group (3612.78?±?468.54) mmHg/s were significantly higher than those of the BMSC group (3245.368?±?468.16) , the -dp/dt of the CLC group (3347.21?±?398.56) mmHg/s were also significantly higher than those of the BMSC group (3014.11?±?412.12) mmHg/s. The myocardial infarction area of the BMSC group and CLC group was reduced obviously compared with the myocardial infarction group, and the myocardial infarction area of CLC group was less than that of the BMSC group (P < 0.01) .

Conclusion

It may be of a better effect that transplanting BMSC induced by IGF-1 into CLC than only transplanting BMSC on myocardial infarction rats.

Key words: Bone marrow, Mesenchymal stem cells, Insulin-like growth factor I, Myocytes, cardiac, Myocardial infarction, Cell transplantation

表1 模型组和假手术组大鼠心功能血流动力学指标变化比较（±s）

分组	动物数	LVSP（mmHg）	LVEDP（mmHg）	+dp/dt（mmHg/s）	-dp/dt（mmHg/s）
假手术组	30	130.42±12.58	2.44±1.11	4087.29±525.17	3865.81±614.37
模型组	40	118.74±11.56	14.29±7.63	2645.17±468.16	2487.80±348.21
t值	?	3.979	-9.605	11.905	11.028
P值	?	< 0.01	< 0.01	< 0.01	< 0.01

表1 模型组和假手术组大鼠心功能血流动力学指标变化比较（±s）

分组	动物数	LVSP（mmHg）	LVEDP（mmHg）	+dp/dt（mmHg/s）	-dp/dt（mmHg/s）
假手术组	30	130.42±12.58	2.44±1.11	4087.29±525.17	3865.81±614.37
模型组	40	118.74±11.56	14.29±7.63	2645.17±468.16	2487.80±348.21
t值	?	3.979	-9.605	11.905	11.028
P值	?	< 0.01	< 0.01	< 0.01	< 0.01

图1 倒置显微镜下观察分离培养的BMSCs（×400）

图2 倒置显微镜下观察BMSC的鉴定（免疫细胞化学染色方法，×400）

表2 IGF-1诱导BMSC不同时间分化为心肌样细胞后α-SMA、Troponin-T和Troponin-I的表达结果（±s）

分组	第3天	第7天	第14天	第21天	F值	P值
α-SMA	1.00±0.00	1.14±0.08	1.58±0.09	1.60±0.05	250.69	< 0.01
Troponin-T	1.00±0.00	1.35±0.06	1.92±0.10	1.85±0.07	445.55	< 0.01
Troponin-I	1.00±0.00	1.17±0.26	1.89±0.08	1.89±0.10	106.36	< 0.01

表2 IGF-1诱导BMSC不同时间分化为心肌样细胞后α-SMA、Troponin-T和Troponin-I的表达结果（±s）

分组	第3天	第7天	第14天	第21天	F值	P值
α-SMA	1.00±0.00	1.14±0.08	1.58±0.09	1.60±0.05	250.69	< 0.01
Troponin-T	1.00±0.00	1.35±0.06	1.92±0.10	1.85±0.07	445.55	< 0.01
Troponin-I	1.00±0.00	1.17±0.26	1.89±0.08	1.89±0.10	106.36	< 0.01

表3 移植BMSC组与CLC组大鼠和心肌梗死组大鼠心功能血流动力学指标变化比较（±s）

分组	动物数	LVSP（mmHg）	LVEDP（mmHg）	+dp/dt（mmHg/s）	-dp/dt（mmHg/s）
心肌梗死组	10	118.74±11.56	14.29±7.63	2645.17±468.16	2487.80±348.21
生理盐水对照组	10	119.67±10.41	14.12±3.89	2754.24±478.25	2498.14±354.34
BMSC组	10	121.25± 9.45	9.15±4.54^a	3245.45±368.76^a	3014.11±412.12^a
CLC组	10	125.14± 6.78	4.67±3.46^ab	3612.78±468.54^ab	3347.21±398.56^ab
F值	?	1.023	42.441	14.367	13.758
P值	?	0.393	< 0.01	< 0.01	< 0.01

表3 移植BMSC组与CLC组大鼠和心肌梗死组大鼠心功能血流动力学指标变化比较（±s）

分组	动物数	LVSP（mmHg）	LVEDP（mmHg）	+dp/dt（mmHg/s）	-dp/dt（mmHg/s）
心肌梗死组	10	118.74±11.56	14.29±7.63	2645.17±468.16	2487.80±348.21
生理盐水对照组	10	119.67±10.41	14.12±3.89	2754.24±478.25	2498.14±354.34
BMSC组	10	121.25± 9.45	9.15±4.54^a	3245.45±368.76^a	3014.11±412.12^a
CLC组	10	125.14± 6.78	4.67±3.46^ab	3612.78±468.54^ab	3347.21±398.56^ab
F值	?	1.023	42.441	14.367	13.758
P值	?	0.393	< 0.01	< 0.01	< 0.01

表4 心肌梗死大鼠注射BMSC与CLC后梗死面积的比较（±s）

分组	动物数	LVSP（mmHg）	LVEDP（mmHg）	+dp/dt（mmHg/s）	-dp/dt（mmHg/s）
心肌梗死组	10	118.74±11.56	14.29±7.63	2645.17±468.16	2487.80±348.21
生理盐水对照组	10	119.67±10.41	14.12±3.89	2754.24±478.25	2498.14±354.34
BMSC组	10	121.25± 9.45	9.15±4.54^a	3245.45±368.76^a	3014.11±412.12^a
CLC组	10	125.14± 6.78	4.67±3.46^ab	3612.78±468.54^ab	3347.21±398.56^ab
F值	?	1.023	42.441	14.367	13.758
P值	?	0.393	< 0.01	< 0.01	< 0.01

表4 心肌梗死大鼠注射BMSC与CLC后梗死面积的比较（±s）

分组	动物数	LVSP（mmHg）	LVEDP（mmHg）	+dp/dt（mmHg/s）	-dp/dt（mmHg/s）
心肌梗死组	10	118.74±11.56	14.29±7.63	2645.17±468.16	2487.80±348.21
生理盐水对照组	10	119.67±10.41	14.12±3.89	2754.24±478.25	2498.14±354.34
BMSC组	10	121.25± 9.45	9.15±4.54^a	3245.45±368.76^a	3014.11±412.12^a
CLC组	10	125.14± 6.78	4.67±3.46^ab	3612.78±468.54^ab	3347.21±398.56^ab
F值	?	1.023	42.441	14.367	13.758
P值	?	0.393	< 0.01	< 0.01	< 0.01

[1]	Hynes B, Kumar AH, O'Sullivan J, et al. Potent endothelial progenitor cell-conditioned media-related anti-apoptotic, cardiotrophic, and pro-angiogenic effects post-myocardial infarction are mediated by insulin-like growth factor-1[J]. Eur Heart J, 2013, 34 (10):782-789.
[2]	van den Akker F, Deddens JC, Doevendans PA, et al. Cardiac stem cell therapy to modulate inflammation upon myocardial infarction[J]. Biochim Biophys Acta, 2013, 1830(2):2449-2458.
[3]	Wang WE, Yang D, Li L, et al. Prolyl hydroxylase domain protein 2 silencing enhances the survival and paracrine function of transplanted adipose-derived stem cells in infarcted myocardium[J]. Circ Res, 2013, 113(3):288-300.
[4]	Ye L, Zhang P, Duval S, et al. Thymosin β4 increases the potency of transplanted mesenchymal stem cells for myocardial repair[J]. Circulation, 2013, 128(11 Suppl 1):S32-41.
[5]	Wang XL, Li CM, Lv Q, et al. Isulin-like growth factor 1 induces bone mesenchymal stem cells differentiation into cardiomyocyte-like cells[J]. Chin J Cardiol, 2013, 41(2):150-155.
[6]	王秀力，刘颖，吕茜, 等. 心力衰竭大鼠骨髓间充质干细胞形态及功能的改变[J]. 中华细胞与干细胞杂志(电子版), 2015, 5(3):185-190.
[7]	Gursu HA, Varan B, Erdogan I. Use of oral budesonide in the management of protein-losing enteropathy due to restrictive cardiomyopathy[J]. Cardiol Young, 2014, 24(4):764-766.
[8]	Yanagawa B, Collazo L, Burton N, et al. Combined heart transplantation and thoracic endovascular aortic repair for heart failure secondary to tricuspid atresia palliated with Potts shunt[J]. Innovations (Phila), 2013, 8(3): 242-244.
[9]	Pelekanos RA, Li J, Gongora M, et al. Comprehensive transcriptome and immunophenotype analysis of renal and cardiac MSC-like populations supports strong congruence with bone marrow MSC despite maintenance of distinct identities[J]. Stem Cell Res, 2012, 8(1): 58-73.
[10]	Zhao Y, Sun X, Cao W, et al. Exosomes derived from human umbilical cord mesenchymal stem cells relieve acute myocardial ischemic injury[J]. Stem Cells Int, 2015, 2015: 761643.
[11]	Yu J, Wu YK, Gu Y, et al. Immuno-modification of enhancing stem cells targeting for myocardial repair[J]. J Cell Mol Med, 2015, 19(7): 1483-1491.
[12]	Jeong H, Yim HW, Cho Y, et al. The effect of rigorous study design in the research of autologous bone marrow-derived mononuclear cell transfer in patients with acute myocardial infarction[J]. Stem Cell Res Ther, 2013, 4(4):82.
[13]	Fisher SA, Dorée C, Brunskill SJ, et al. Bone marrow stem cell treatment for ischemic heart disease in patients with no option of revascularization: a systematic review and meta-analysis[J]. PLoS One, 2013, 8(6):e64669.
[14]	Williams AR, Suncion VY, McCall F, et al. Durable scar size reduction due to allogeneic mesenchymal stem cell therapy regulates whole-chamber remodeling[J]. J Am Heart Assoc, 2013, 2(3):e000140.
[15]	Liu C, Fan Y, Zhou L, et al. Pretreatment of mesenchymal stem cells with angiotensin II enhances paracrine effects, angiogenesis, gap junction formation and therapeutic efficacy for myocardial infarction[J]. Int J Cardiol, 2015, 188:22-32.
[16]	Rong XS, Li MQ, Xu Y, et al. Injection of cardiac stem cells prolongs the survival of cardiac allograft rats[J]. Am J Med Sci, 2015, 349(1): 67-71.

[1]	陈慧, 姚静, 张宁, 刘磊, 马秀玲, 王小贤, 方爱娟, 管静静. 超声心动图在多发性骨髓瘤心脏淀粉样变中的诊断价值[J/OL]. 中华医学超声杂志（电子版）, 2024, 21(10): 943-949.
[2]	何甘霖, 陈香侬, 李萍, 甄佳怡, 李京霞, 邹外一, 许多荣. 白血病异基因造血干细胞移植术后股骨坏死的影响因素[J/OL]. 中华关节外科杂志(电子版), 2024, 18(04): 450-456.
[3]	王友芳, 李兴超, 刘清敏, 刘德彬, 刘松伍, 郭冬冬, 车峰远. 应激性高血糖指数对经皮冠状动脉介入术后急性心肌梗死患者发生主要不良心脑血管事件的预测价值[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(02): 124-129.
[4]	徐志刚, 曹涛, 何亭, 李博奥, 魏婧韬, 张栋梁, 官浩, 杨薛康. 采用抗生素骨水泥治疗糖尿病患者心脏术后胸骨骨髓炎的临床效果观察[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(06): 498-502.
[5]	罗文斌, 韩玮. 胰腺癌患者首次化疗后中重度骨髓抑制的相关危险因素分析及预测模型构建[J/OL]. 中华普通外科学文献(电子版), 2024, 18(05): 357-362.
[6]	刘云, 时月, 郭冬梅, 邱志远, 王丽娟, 冉学红, 李乾鹏. 造血干细胞移植治疗伴有胚系突变的髓系肿瘤患者三例并文献复习[J/OL]. 中华移植杂志(电子版), 2024, 18(04): 230-234.
[7]	王向丽, 吴涛, 毛东锋, 刘恒, 刘文慧, 周芮, 田红娟. 异基因造血干细胞移植治疗ANKRD26相关性血小板减少症1例并文献复习[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 236-238.
[8]	杨阳, 王琤, 周文土, 周冰. Caveolae/Caveolin-1与膜胆固醇共同调控小鼠BMSCs成骨分化[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 137-142.
[9]	汪鹏飞, 程莹莹, 赵海康. 骨髓间充质干细胞改善神经病理性疼痛的机制探讨[J/OL]. 中华脑科疾病与康复杂志(电子版), 2024, 14(04): 230-234.
[10]	李忠清, 罗军. 尿蛋白电泳在浆细胞疾病鉴别诊断中的价值[J/OL]. 中华诊断学电子杂志, 2024, 12(04): 217-221.
[11]	刘聪辉, 何浩然, 黄一诺, 张凤, 王凡月, 郝翰. 膳食铜补充对大鼠心肌梗死后心肌基质金属蛋白酶2表达水平及血流动力学的影响[J/OL]. 中华诊断学电子杂志, 2024, 12(03): 166-172.
[12]	张滕, 陶艳玲. Shwachman-Diamond综合征继发骨髓增生异常综合征一例及文献复习[J/OL]. 中华诊断学电子杂志, 2024, 12(03): 178-182.
[13]	徐来英, 程效, 戴亨纷, 侯俊凉, 苏怡林, 张彦. 药物联合个体化精准恒定功率运动疗法治疗心肌梗死术后频发室性早搏一例[J/OL]. 中华心脏与心律电子杂志, 2024, 12(03): 176-179.
[14]	蔡昊旻, 郭亮, 章靖, 周逸鸣, 罗恒. 胸壁曲霉菌感染3 例报告[J/OL]. 中华胸部外科电子杂志, 2024, 11(04): 242-245.
[15]	郑屹, 刘莹, 张煜坤, 李广平, 陈康寅, 刘彤. 既往及新发心房颤动对急性心肌梗死患者远期卒中风险的影响[J/OL]. 中华脑血管病杂志(电子版), 2024, 18(05): 406-417.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Curative effect of transplantion of cardiomyocyte-like cells on myocardial infarction rats

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Curative effect of transplantion of cardiomyocyte-like cells on myocardial infarction rats