主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2020, Vol. 43 ›› Issue (2): 115-124.doi: 10.3969/j.issn.1006-2157.2020.02.005

• 中药药理 • 上一篇    下一篇

补阳还五汤联合BMSCs对心肌缺血再灌注损伤模型大鼠心肌的防护作用*

宋忠阳1, 王功臣2, 张志明2#, 雍文兴3, 刘永琦4, 张利英4, 李娟3   

  1. 1 甘肃中医药大学中医临床学院 甘肃 730000;
    2 甘肃中医药大学附属医院肿瘤科;
    3 甘肃中医药大学附属医院急诊科;
    4 甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室
  • 收稿日期:2019-09-06 发布日期:2020-03-24
  • 通讯作者: 张志明,男,主任医师,教授,博士生导师,研究方向:急危重症的中医药防治,E-mail:zhangzhimingys@163.com
  • 作者简介:宋忠阳,男,在读博士生
  • 基金资助:
    *国家自然科学基金资助项目(No.81660730),甘肃省重点研发计划(No.18YF1FA044)

Buyang Huanwu Decoction’s protective effect on myocardium and influence on BMSCs migration in myocardial ischemia/reperfusion rats*

Song Zhongyang1, Wang Gongchen2, Zhang Zhiming2*, Yong Wenxing3, Liu Yongqi4,Zhang Liying4, Li Juan3   

  1. 1 Clinical College of Chinese Medicine, Gansu University of Chinese Medicine;
    2 Department of Oncology, Affiliated Hospital of Gansu University of Chinese Medicine;
    3 Emergency Department, Affiliated Hospital of Gansu University of Chinese Medicine;
    4 Provincial-Level Key Laboratory for Molecular Medicine of Major Diseases and The Prevention and Treatment with Traditional Chinese Medicine Research in Gansu Colleges and Universities
  • Received:2019-09-06 Published:2020-03-24
  • Contact: Prof. Zhang Zhiming, Chief Physician. Affiliated Hospital of Gansu University of traditional Chinese Medicine, Gansu 730000, E-mail: zhangzhimingys@163.com
  • Supported by:
    National Natural Science Foundation of China (No. 81660730); Key Research and Development Plan of Gansu Province (No. 18YF1FA044)

摘要: 目的 研究补阳还五汤联合骨髓间充质干细胞(BMSCs)对心肌缺血再灌注损伤(MIRI)大鼠心肌的防护作用及BMSCs迁移的的影响。方法 60只Wistar大鼠随机分为假手术组、模型组、干细胞组、补阳还五汤+干细胞组,结扎冠状动脉左前降支(LAD)方法复制MIRI大鼠模型,分别采用生理盐水和补阳还五汤灌胃14 d。生化法检测血清肌钙蛋白I(cTnI)、肌酸激酶同工酶(CK-MB)及乳酸脱氢酶(LDH)水平,血清抗氧化因子丙二醛(MDA)浓度,血清超氧化物歧化酶(SOD)活性情况及一氧化氮(NO)含量。超声心动图检测大鼠心功能,HE染色检测心肌组织病理形态,小动物活体成像系统观察荧光GFP蛋白标记的BMSCs在体内移行分布情况,免疫荧光检测心肌组织BMSCs含量。结果 与干细胞组比较,补阳还五汤+干细胞组大鼠左室舒张末经(LVIDd)和左室收缩末经(LVIDs) 显著减小、而射血分数(EF)增加(P<0.05);血清cTnI、LDH、CK-MB水平均明显降低(P<0.05);血清SOD活性、NO含量均升高,MDA含量均降低(P<0.05)。HE染色结果显示,补阳还五汤+干细胞组大鼠心肌细胞结构恢复正常,肌纤维排列整齐,细胞间质有少量水肿,有少量的炎症细胞浸润,但细胞界限清楚。小动物活体成像系统显示,假手术组和模型组无明显 GFP蛋白BMSCs移行分布,补阳还五汤+干细胞组GFP表达强度明显的强于干细胞组,且心脏部位BMSCs的浓度显著高于干细胞组(P<0.05)。免疫荧光检测结果显示, 补阳还五汤+干细胞组大鼠心肌组织BMSCs含量明显高于干细胞组(P<0.05)。结论 补阳还五汤能减轻移植BMSCs大鼠心肌缺血再灌注后损伤,对心肌具有保护作用,其机制与减轻细胞膜脂质过氧化反应,防护再灌注期心肌细胞的氧化应激损伤有关,同时补阳还五汤能够促进BMSCs向MIRI模型大鼠心肌组织迁移,提高BMSCs的归巢效率。

关键词: 补阳还五汤, 骨髓间充质干细胞, 心肌缺血再灌注损伤, 防护作用, 迁移, 大鼠

Abstract: Objective To study the protective effect of Buyang Huanwu Decoction (BHD) combined with bone mesenchymal stem cells (BMSCs) on myocardial ischemia reperfusion injury (MIRI) in rats and the effect of BMSCs migration. Methods 60 Wistar rats were randomly divided into sham operated group, model group, BMSCs group and BHD plus BMSCs group The left anterior descending coronary artery (LAD) was ligated to duplicate Miri rat model in all groups except for the sham operated group. The rats were gavaged with BHD and normal saline for 14 days respectively. The levels of cTnI, CK-MB, LDH,and MDA, the content of nitric oxide (NO) and the activity of SOD in serum were measured by biochemical method. The cardiac function was detected by echocardiography, the pathological morphology of myocardial tissue was detected by HE staining, the migration and distribution of GFP-labeled BMSCs were observed in vivo by Caliper IVIS Lumina II, and the content of myocardial BMSC was detected by immunofluorescence. Results Compared with the BMSCs group, in BHD plus BMSCs group, the LVIDd and LVIDs decreased significantly, while EF was increased (P<0.05); the levels of cTnI, LDH and CK-MB were significantly decresed (P<0.05); the activity of SOD and the content of NO were increased, while the content of MDA was decreased (P<0.05). According to HE staining, the muscle fibers were arranged in order; there was slight edema in the intercellular matrix and a little infiltration of inflammatory cells, but the cell boundary was clear. The Caliper IVIS Lumina II in vivo showed that there were no obvious migration and distribution of GFP-labeled BMSCs in the sham operation group and model group, but the expression intensity of GFP in the BHD plus BMSCs group was significantly higher than that in the BMSCs group, and the concentration of BMSCs in the heart was significantly higher than that in the BMSCs group (P<0.05). The results of immunofluorescence showed that the content of BMSCs in the BHD plus BMSCs group was significantly higher than that in the BMSCs group (P<0.05). Conclusions BHD could possibly reduce transplanted BMSCs after myocardial ischemia reperfusion injury in rats, reduce myocardial tissue pathological morphological changes, improve cardiac function. Through improving SOD activity in serum, increasing the NO content, and reducing the CK-MB, LDH, cTnI levels and MDA concentration in serum, it seems to reduce MIRI with the myocardial protective effect. Its mechanism might be related to the reduction of the membrane lipid peroxidation, and protection from the oxidative stress injury of myocardial cells in reperfusion period. It also indicates that BHD could possibly promote the migration of BMSCs into the myocardial tissue of MIRI model rats, increase the number of BMSCs migration, and improve the homing efficiency of BMSCs.

Key words: Buyang Huanwu Decoction (BHD), bone marrow mesenchymal stem cells, myocardial ischemia-reperfusion injury, protective effect, migration, rats

中图分类号: 

  • R285.5