蒺藜皂苷对阿霉素损伤心肌细胞的保护作用

Effect of gross saponins of Tribulus terrestris on cardiocytes impaired by adriamycin

观察蒺藜皂苷(gross saponins of Tribulus terrestris,GSTT)对阿霉素(adriamycin,ADR)诱导大鼠乳鼠心肌细胞损伤的保护作用,并初步探讨其作用机制。分离出生1～3d大鼠心肌细胞,培养72h后随机分为正常对照组、ADR(终浓度为2.0mg·L-1)模型组和GSTT(100、30及10mg·L-1)组,继续培养24h后,应用MTT比色法检测细胞存活率,测定培养基中肌酸激酶(CK)、乳酸脱氢酶(LDH)、天门冬氨酸氨基转移酶(AST)释放量,超氧化物歧化酶(SOD)活性,丙二醛(MDA)、一氧化氮(NO)含量,并采用流式细胞仪检测细胞凋亡及应用Western blotting检测GSTT对凋亡相关蛋白caspase-3的作用。结果表明,与正常对照组比较,ADR模型组心肌细胞存活数明显减少,心肌细胞培养液中CK、LDH、AST释放量增加(P<0.01,P<0.001),同时SOD活力下降(P<0.01)而MDA、NO含量升高(P<0.001);与ADR模型组比较,GSTT(100和30mg·L-1)组存活心肌细胞数增多(P<0.05,P<0.001),心肌细胞培养液中CK、LDH、AST含量明显降低,SOD活力增加、MDA和NO含量降低(P<0.05,P<0.01,P<0.001)。流式细胞仪检测GSTT(100和30mg·L-1)组心肌细胞凋亡数明显减少(P<0.05,P<0.01),caspase-3含量下降(P<0.05,P<0.001)。GSTT对ADR损伤的心肌细胞具有保护作用,可减轻心肌细胞损伤,抑制心肌细胞凋亡,其机制与抗自由基作用有关。

This study is to observe the protection of gross saponins of Tribulus terrestris （GSTT） on cardiocytes impaired by adriamycin （ADR） and approach its mechanism of action. Cardiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, model （ADR 2 mg·L^-1） group, and GSTT （100, 30, and 10 mg·L^-1） groups. MTT colorimetric method was deployed to detect cardiocyte survival rate, activities of CK, LDH, AST, SOD, MDA and NO were detected, and apoptosis was detected with flow cytometry. Effect of GSTT on caspase-3 was detected with Western blotting. Compared with control group, contents of CK, LDH, AST, MDA and NO were increased, and activity of SOD was reduced （P〈0.05, P〈0.01, P〈0.001） by ADR. Numbers of survival cells were increased （P〈0.05, P〈0.001）, contents of CK, LDH, AST, MDA and NO were decreased, and activity of SOD was increased （P〈0.05, P〈0.01, P〈0.001） by GSTT （100 and 30 mg·L^-1）. Apoptosis of cardiocytes and concentration of caspase-3 can be reduced by GSTT （100 and 30 mg·L^-1）. GSTT can protect cardiocytes impaired by ADR, which are possible involved with its effect of resisting oxygen free radical.