丹参素冰片酯对同型半胱氨酸诱导大鼠骨髓间充质干细胞损伤的保护作用及机制  

Protective effect and related mechanism of tanshinol borneol ester on homocysteine induced rat bone marrow mesenchymal stem cells damage

作　　者：王雪[1] 杨婧[3] 袁红[2] 刘晓丽[2] 李艳[2] 贾璞[4] 郑晓晖[4] 郑小璞[2] 

机构地区：[1]西安交通大学第一附属医院心血管外科,710061 [2]西安交通大学第一附属医院心血管内科,710061 [3]西安市第四医院重症医学科 [4]西北大学中草药现代化研究及工程中心西北大学生命科学学院

出　　处：《中华心血管病杂志》2017年第2期130-136,共7页Chinese Journal of Cardiology

摘　　要：目的探讨丹参素冰片酯（TBE）对同型半胱氨酸（Hcy）诱导大鼠骨髓间充质干细胞（BMSC）损伤的作用及机制。方法以密度梯度离心法结合贴壁培养法体外分离和培养BMSC。将BMSC分为对照组（未干预）、TBE-1组（加入含10 μmol/L TBE-1溶液100 μl）、TBE-2组（加入含10 μmol/L TBE-2溶液100 μl）、Hcy组（加入含0.5 mmol/L Hcy溶液100 μl）、Hcy＋TBE-1组（加入含10 μmol/L TBE-1溶液100 μl和含0.5 mmol/L Hcy溶液100 μl）、Hcy＋TBE-2组（加入含10 μmol/L TBE-2溶液100 μl和含0.5 mmol/L Hcy溶液100 μl）、Hcy＋TBE-1＋阻断剂组[加入含10 μmol/L TBE-1溶液100 μl、0.5 mmol/L Hcy溶液100 μl和25 μmol/L LY294002（磷脂酰肌醇3激酶的特异性阻断剂）溶液100 μl]和Hcy＋TBE-2＋阻断剂组（加入含10 μmol/L TBE-2溶液100 μl、0.5 mmol/L Hcy溶液100 μl和25 μmol/L LY294002溶液100 μl）。采用四甲基偶氮唑盐（MTT）法检测细胞增殖活性；黄嘌呤氧化酶法检查细胞总超氧化物歧化酶（T-SOD）活性，TBA法检测细胞丙二醛水平，评价细胞氧化损伤程度；透射电子显微镜观察各组细胞超微结构；免疫细胞化学法检测细胞蛋白激酶B（PKB）及核转录因子-κB（NF-κB）的表达水平。结果（1）干预1、12、24和48 h后，TBE-1组和TBE-2组细胞的增殖活性均高于正常组（P均〈0.01），TBE-1组与TBE-2组的细胞增殖活性差异均无统计学意义（P均〉0.05）。（2）TBE-1组和TBE-2组的T-SOD活性均高于对照组（P均〈0.01）；Hcy组、Hcy＋TBE-1组和Hcy＋TBE-2组的T-SOD活性均低于对照组（P均〈0.01）；对照组、Hcy＋TBE-1＋阻断剂组和Hcy＋TBE-2＋阻断剂组之间的T-SOD活性差异均无统计学意义（P均〉0.05）；Hcy＋TBE-1组和Hcy＋TBE-2组的T-SOD活性均高于Hcy组（P均〈0.01）；Hcy＋TBE-1＋阻断剂组的T-SOD活性高于Hcy＋TBE-1组（P〈0.05）；Hcy＋TBE-2＋阻断剂组的T-SOD活性高于Hcy＋TBE-2组（P〈0.05）�ObjectiveTo investigate the protective effect and potential mechanism of tanshinol borneol ester （TBE） on homocysteine（Hcy） induced rat bone marrow mesenchymal stem cells （BMSCs） damage. MethodsBMSCs were isolated and cultured in vitro by density gradient centrifugation and adherent culture method. BMSCs were divided into the control （normal isolation and culture）, TBE-1（10 μmol/L TBE-1 solution with 100 μl）, TBE-2 （10 μmol/L TBE-2 solution with 100 μl）, Hcy （0.5 mmol/L Hcy solution with 100 μl）, Hcy ＋ TBE-1（0.5 mmol/L Hcy solution with 100 μl, and 10 μmol/L TBE-1 solution with 100 μl）, Hcy ＋ TBE-2 （0.5 mmol/L Hcy solution with 100 μl, and 10 μmol/L TBE-2 solution with 100 μl）, Hcy＋ TBE-1＋ inhibitor group（0.5 mmol/L Hcy solution with 100 μl, 10 μmol/L TBE-1 solution with 100 μl, and 25 μmol/L LY294002（specific blocker of phosphatidylinositol 3 kinase） solution with 100 μl）, Hcy＋ TBE-2＋ inhibitor group（0.5 mmol/L Hcy solution with 100 μl, 10 μmol/L TBE-2 solution with 100 μl, and 25 μmol/L LY294002 solution with 100 μl）. Cell proliferation activity was detected by MTT assay. The T-SOD activity and malonaldehyde level of cells were measured by anthineoxidase method and TBA method, respectively, to evaluate cell oxidative and antioxidative activities. The ultrastructure of cells was observed under transmission electron microscope. The expression level of PKB and NF-κB of cells in various groups were detected with the immunocytochemical method. Results（1）Cell proliferation activity in TBE-1 group and TBE-2 group was significantly increased compared with the control group （both P〈0.01）, and was similar between TBE-1 group and TBE-2 groups after 1, 12, 24 and 48 hours treatment.（2）The T-SOD activity in TBE-1 group and TBE-2 group was significantly higher than in control group （both P〈0.01）, while it was significantly lower in Hcy group, Hcy＋ TBE-1 group, and Hcy＋ TBE-2 group than in control group（all P〈0.01�

关 键 词：间质干细胞 高半胱氨酸 丹参素冰片酯 

分 类 号：R965[医药卫生—药理学]