蒺藜皂苷单体B对心肌细胞缺氧/复氧损伤的保护作用  

Protective effect of Tribulus terrestris saponin monomer B on cadiocytes impaired by hypoxia-reoxygenation

作　　者：张爽[1] 李红[1] 杨世杰[1] 

机构地区：[1]吉林大学白求恩医学院药理学系,吉林长春130021

出　　处：《中国药理学通报》2010年第2期208-212,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30472020;30672654);高校博士点基金资助项目(No2005183129)

摘　　要：目的观察蒺藜皂苷单体B(TTSMB)对大乳鼠心肌细胞缺氧/复氧损伤的保护作用。方法分离出生1～3d大鼠心肌细胞,培养72h后随机分为:正常对照组(Control)、缺氧/复氧模型组(H/R)、蒺藜皂苷组(GSTT)、蒺藜皂苷单体B(TTSMB)10、1、0.1nmol·L-1组。观察细胞形态学变化,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)含量,并采用流式细胞仪检测心肌细胞凋亡率及应用Western blot检测TTSMB对凋亡相关蛋白caspase-3的作用。结果与对照组比较,H/R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),SOD活力下降,MDA含量升高(P<0.01);与模型组比较,TTSMB10、1、0.1nmol·L-1组存活心肌细胞数明显增多(P<0.05,P<0.01),心肌细胞培养液中CK、LDH、AST含量降低,SOD活力增加、MDA含量降低(P<0.05,P<0.01),心肌细胞凋亡率明显下降,caspase-3表达亦下降。结论TTSMB可减轻心肌细胞缺氧/复氧损伤,抑制心肌细胞凋亡,具有心肌细胞保护作用,其机制与抗自由基作用有关。Aim To observe the protection effect of Tribulus terrestris saponin monomer B （TTSMB） on cadiocytes impaired by hypoxia-reoxygenation （H/R）. Methods Cadiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, H/ R group, GSTT 100 mg · L^-1 group and TTSMB 10,1, 0. 1 nmol · L^-1 group. Morphocytology change of cadiocytes was observed after the treatment. Cadiocyte survival rate was detected with MTT colorimetric method. Levels of creatine kinase （CK）, lactate dehydrogenase （ LDH）, aspartate aminotransferase （AST） , malondialdehyde （MDA）, and activity of superoxide dismutase （SOD） were determined. Apoptosis rate was detected with flow cytometry. Expression of caspase-3 was examined with western blot. Results Compared with the control group, the survival cell number in H/R group decreased obviously, content of CK, LDH, AST, MAD increased, and activity of SOD decreased （P 〈 0. 01 ）. Compared with the model group, the survival cell population increased in TFSMB 10, 1,0. 1 nmol · L^-1 group （P 〈0. 05 and P 〈0. 01 ）. Contents of CK, LDH, AST, MAD decreased, whereas the activity of SOD increased （P 〈0.05 and P 〈0. 01 ）. Apoptosis rate and expression of caspase-3 also reduced in TTSMB 10, 1,0. 1 nmol · L^-1 group. Conclusion TTSMB has significant cadiocytes protective effect against H/R and inhibits cadiocyte apoptosis, and the mechanism depends on the effect against oxygen free radical.

关 键 词：蒺藜皂苷单体B 心肌细胞 缺氧/复氧损伤(H/R) 凋亡 caspase-3 氧自由基 

分 类 号：R-332[医药卫生] R284.1