| 引用本文: | 沈云方,费莹,王建琴,秦宇芬.丹参多酚酸盐协同间充质干细胞自体移植治疗大鼠急性心肌损伤[J].中国现代应用药学,2021,38(8):944-952. |
| SHEN Yunfang,FEI Ying,WANG Jianqin,QIN Yufen.Study on Salvianolate Combined with Mesenchymal Stem Cells Autotransplantation in the Treatment of Acute Myocardial Injury[J].Chin J Mod Appl Pharm(中国现代应用药学),2021,38(8):944-952. |
|
| 摘要: |
| 目的 研究丹参多酚酸盐协同间充质干细胞（mesenchymal stem cells，MSCs）自体移植治疗急性心肌损伤（acutemyocardial injury，AMI）的效果。方法 分离SD大鼠骨髓MSCs，体外培养，并加丹参多酚酸盐进行诱导干预（100 mg·L-1）。采用免疫荧光检测MSCs中cTnI、NKx2.5、GATA-4的荧光表达，Western blotting检测心肌细胞标志物cTnI、CX43、NKx2.5、GATA-4、β-MHC的蛋白表达情况。制备AMI动物模型，SD大鼠随机分为假手术组（PBS+生理盐水）、模型组（PBS+生理盐水）、MSCs组（60 μL MSCs+生理盐水）、丹参多酚酸盐组（PBS+30 mg·kg-1丹参多酚酸盐）以及丹参多酚酸盐+MSCs组（60 μL MSCs+30 mg·kg-1丹参多酚酸盐）。AMI模型制备后连续给药7 d，每天1次。7 d后记录各组大鼠心功能参数，随后颈动脉采血，并处死大鼠采集心脏组织样本。采用TTC染色测定心肌梗死面积，ELISA检测肌酸激酶MB同工酶（CK-MB）、乳酸脱氢酶（LDH）活性，HE染色观察心肌组织病理损伤情况，TUNEL染色检测心肌组织细胞凋亡情况。结果 成功分离培养骨髓MSCs，经丹参多酚酸盐诱导干预后，免疫荧光检测显示cTnI、NKx2.5、GATA-4荧光表达明显增强，Western blotting检测显示cTnI、CX43、NKx2.5、GATA-4、β-MHC蛋白表达显著提高。成功制备AMI大鼠模型并给予药物处理后，结果显示与模型组相比，丹参多酚酸盐+MSCs组的心肌功能参数FS、+dp/dtmax、-dp/dtmax水平显著上升，LVDEP水平降低，CK-MB、LDH活性水平显著降低（P<0.01或P<0.05）。TTC染色结果显示各给药组大鼠心肌梗死面积均极显著降低，以丹参多酚酸盐+MSCs组降低最为明显（P<0.01）。HE染色及评分结果显示丹参多酚酸盐+MSCs联合处理能有效改善AMI模型大鼠的心肌损伤。TUNEL染色结果显示丹参多酚酸盐+MSCs组阳性细胞数减少且着色较浅，细胞凋亡程度改善最明显。结论 丹参多酚酸盐协同MSCs自体移植可有效治疗AMI，其作用机制可能与丹参多酚酸盐定向诱导MSCs分化为心肌样细胞，改善血流动力学，降低心肌酶活性，抑制细胞凋亡等有关。 |
| 关键词: 急性心肌损伤 丹参多酚酸盐 MSCs 细胞凋亡 |
| DOI：10.13748/j.cnki.issn1007-7693.2021.08.008 |
| 分类号:R965.2 |
| 基金项目:浙江省湖州市科技技术局项目(2017GY44) |
|
| Study on Salvianolate Combined with Mesenchymal Stem Cells Autotransplantation in the Treatment of Acute Myocardial Injury |
|
SHEN Yunfang, FEI Ying, WANG Jianqin, QIN Yufen
|
|
Department of Pharmacy, Huzhou Hospital of Traditional Chinese Medicine Affiliated to Zhejiang University of Traditional Chinese Medicine, Huzhou 313000, China
|
| Abstract: |
| OBJECTIVE To investigate the effect of salvianolate combined with mesenchymal stem cells(MSCs) autotransplantation in the treatment of acute myocardial injury(AMI). METHODS MSCs were isolated from bone marrow of SD rats, cultured in the medium and incubated by salvianolate(100 mg·L-1). Immunofluorescence was used to detect the fluorescence expression of cTnI, NKx2.5 and GATA-4 in MSCs, Western blotting was used to detect the expression of cTnI, Cx43, Nkx2.5, GATA-4 and β-MHC. The animal model of AMI was prepared, and SD rats were randomly divided into five groups:sham operation group(PBS + saline), model group(PBS + saline), MSCs group(60 μL MSCs + saline), salvianolate group(PBS + 30 mg·kg-1 salvianolate) and salvianolate + MSCs group (60 μL MSCs + 30 mg·kg-1 salvianolate). After AMI model was established, SD rats in each group were administrated consecutively for seven days, once per day. And then the heart function parameters of each group were recorded, and the blood samples and heart tissue samples were collected after seven days. The myocardial infarction area was measured by TTC staining, the activity of CK-MB and LDH were detected by ELISA, HE staining was used to assess the pathological injury of myocardial tissue and TUNEL staining was used to detect the apoptosis of myocardial cells. RESULTS MSCs were successfully isolated and cultured, and with salvianolate induced, immunofluorescence results showed that the expression of cTnI, Nkx2.5 and GATA-4 increased significantly, Western blotting showed that the expression of cTnI, CX43, Nkx2.5, GATA-4 and β-MHC was significantly increased. After the AMI rat model was successfully prepared and treated, the results showed that compared with the model group, the levels of myocardial function parameters including FS, +dp/dtmax and -dp/dtmax in salvianolate + MSCs group increased significantly, while the levels of LVDEP and the CK-MB and LDH activity decreased significantly(P<0.01 or P<0.05). The results of TTC staining showed that compared with the model group, the myocardial infarct area of rats in each administration group decreased significantly, especially in the group of salvianolate + MSCs (P<0.01). HE staining results showed that the combination of salvianolate and MSCs could effectively alleviate the myocardial injury in AMI rats. TUNEL staining results showed that the number of positive cells in salvianolate + MSCs group was decreased and the degree of apoptosis decreased most obviously. CONCLUSION Salvianolate and MSCs autotransplantation can effectively alleviated AMI, this might be related to the differentiation of MSCs into cardiomyocytes, the improvement of hemodynamics, the reduction of myocardial enzyme activity, and the inhibition of apoptosis. |
| Key words: acute myocardial injury salvianolate MSCs apoptosis |
