丹参酚酸B抑制TLR4-NFκB-TNFα炎症损伤通路防治心肌细胞的缺氧损伤  

Salvianolic Acid Binhibits TLR4-NFκB-TNFα Pathway and Attenuates Neonatal rat Cardiomyocytes Damage Induced by Hypoxia

作　　者：张云[1] 王阶[1,2] 吴广均[1] 刘瑞华[1] 

机构地区：[1]中国中医科学院广安门医院,北京100053 [2]湖北中医药大学,湖北武汉430065

出　　处：《辽宁中医杂志》2012年第1期20-22,共3页Liaoning Journal of Traditional Chinese Medicine

基　　金：广安门医院科研基金(81359)

摘　　要：目的:探讨TLR4-NFκB-TNFα损伤通路和HSP70在体外缺氧培养心肌细胞中的变化规律以及丹参酚酸B的调节作用。方法:分离纯化出生1～3天Wistar乳鼠心肌细胞并复制体外缺氧培养心肌细胞模型。丹参酚酸B(SalB)大、中、小剂量浓度分别为10-5mol.L-1、10-6mol.L-1和10-7mol.L-1并在缺氧培养前6h进行干预。细胞爬片免疫组化法检测tool样受体4(TLR4)和核因子κB(NFκB)蛋白的表达,ELISA法检测细胞上清液中肿瘤坏死因子α(TNFα)和热休克蛋白70(HSP70)的浓度,分光光度计法检测细胞上清液中丙二醛(MDA)和乳酸脱氢酶(LDH)的含量。结果:与正常组比较,缺氧培养6h后心肌细胞上清液中MDA、LDH、TNFα含量均显著升高(3.71±0.84)VS(1.78±0.56)(,561.41±129.31)VS(36.80±10.22)(,89.39±21.65)VS(49.42±4.16),P<0.055或P<0.051);心肌细胞TLR4和NFκB蛋白的表达量也显著升高(P<0.055或P<0.051)。与单纯缺氧培养组比较,丹参酚酸B大剂量组MDA含量显著下降(1.94±0.49)VS(3.71±0.84),P<0.051;大、中、小剂量组LDH含量均显著下降(P<0.051);大、中剂量组TNFα含量显著下降(P<0.055);丹参酚酸B大、中、小剂量组NFκB及TLR4蛋白的表达面积或强度均有明显的下降(P<0.055或P<0.051)。结论:TLR4-NFκB-TNFα炎症通路在缺氧培养6h后即明显激活,该通路的激活在缺氧损伤初期(6h内)是不依赖HSP70存在的。丹参酚酸B预防给药对缺氧培养的体外心肌细胞炎症损伤有明显的保护作用。该作用可能与抑制TLR4-NFκB-TNFα炎症通路有关。Objective：To investigate the change of TLR4-NFκB-TNFα pathway and the possible protect effects and mechanism of Salvianolic acid B on hypoxia model of neonatal rat cardiomyocytes in vitro. Methods：l -3d neonatal Wistar rat was used to isolate cardiomyocytes. The hypoxia model of neonatal rat cardiomyocytes was established by hypoxia chamber. Salvianolic acid B high,mid and low dose（ 10-5mol · L-1 , 10 -6mol · L-1 , 10-7mol· L-1 ） was pre - treated 6h before hypoxia - inducing. The protein expression of tool like receptor 4 （TLR4） and nucleuar factor kappa B （NFKB） was detected by immunohistochemistry . The concentration of tumor necros factorα（TNFα）and heat shock protein 70 （HSP70） in supernatant was detected by ELISA essay and the concentration of malondialdehyde（MDA） and lactate dehydrogenase （LDH） in supernatant was detected by spectro- photometer. Results：Compared with normal group, the concentration of MDA, LDH and TNFet in supernatant increased significantly in hypoxia group （3.71 ± 0. 84 ） VS （ 1.78 ± 0. 56 ）, （ 561.41 ± 129.31 ） VS （ 36. 80 ± 10. 22 ）, （ 89.39 ± 21.65 ） VS （ 49.42 ± 4. 16）, P 〈0. 055 or P 〈0. 051 ）. The expression intensity of NFKB and TLR4 protein in cardiomyocytes increased significantly in hypoxia group （P 〈0. 055 or P 〈0. 051 ）. Compared with hypoxia group,the concentration of MDA decreased significantly in Salvianolic acid B high dose pre - treat group （ 1.94± 0.49 ） VS （ 3.71 ± 0.84, P 〈 0.051 ）. The concentration of LDH decreased significantly in Salvianolic acid B high, mid and low dose group（ P 〈 0. 051 ）. The concentration of TNFa decreased significantly in Salvianolic acid B high and mid dose groUp（ P 〈 0. 055 ）. The expression area or intensity of NFKB and TLR4 protein decreased significantly in Salvianolic acid B high, mid and low dose group （ P 〈 0. 055 or P 〈 0. 051 ）. Conclusion ： TLR4-NFκB-TNFα pathway was activated quickly on hypoxia cardiomyocytes and

关 键 词：丹参酚酸B 缺氧 心肌细胞 TLR4-NFκB-TNFα通路 

分 类 号：R285.5[医药卫生—中药学]