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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。
To observe the protective effect of gross saponins of Tribulus terrestris (GSTT) on cardiomyocyte injury induced by adriamycin (ADR) in rats, and to explore its possible mechanism. The cardiomyocytes of 1 ~ 3d rats were isolated and cultured for 72h. They were randomly divided into normal control group, model group with ADR (2.0mg · L-1) and GSTT group (100,30 and 10mg · L-1) After cultured for 24h, the cell viability was detected by MTT colorimetric assay. The release of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), superoxide dismutase (SOD) activity, malondialdehyde (MDA) and nitric oxide (NO) contents were detected by flow cytometry. Apoptosis was detected by flow cytometry. The effect of GSTT on apoptosis-related protein caspase-3 was detected by Western blotting. The results showed that compared with the normal control group, the survival rate of cardiomyocytes in ADR model group decreased significantly, and the release of CK, LDH and AST in cardiomyocyte culture medium increased (P <0.01, P <0.001) (P <0.001). Compared with ADR model group, the number of viable cardiomyocytes in GSTT group (100 and 30 mg · L -1) increased (P <0.05, P <0.001) The levels of CK, LDH and AST were significantly decreased, the activity of SOD increased, the content of MDA and NO decreased (P <0.05, P <0.01, P <0.001). The number of apoptotic cardiomyocytes in GSTT (100 and 30 mg · L-1) groups was significantly decreased by flow cytometry (P <0.05, P <0.01), while the caspase-3 content was decreased (P <0.05, P <0.001). GSTT has protective effect on ADR-injured cardiomyocytes, which can reduce cardiomyocyte injury and inhibit cardiomyocyte apoptosis, and its mechanism is related to anti-free radical effect.