粉防己碱对缺氧/复氧乳鼠心肌细胞前炎症因子的影响

Effect of tetrandrine on anoxiareoxygenation-induced release of proinflammatory factors in cultured cardiocyte of neonate rats

目的:研究粉防己碱对离体培养的大鼠心肌细胞在缺氧/复氧条件下对前炎症因子:肿瘤坏死因子(TNF-α)、白细胞介素-1(IL-1β)、白细胞介素-6(IL-6)的影响。方法:乳鼠心肌细胞体外培养心肌细胞成功后,随机分为三组:对照组、缺氧/复氧(A/R)组、粉防己碱(Tet)组。对照组:不进行A/R处理,正常情况下持续培养26h;A/R组:在高纯度氩气,无糖无血清培养基条件下培养2h,复氧开始时加入0.9%盐水,再复氧培养24h;Tet组:预先给予终浓度为30μmol/L的Tet孵育60 min,再在无血清低糖DMEM培养基缺氧孵育2h,然后继续复氧培养24h;检测各组在复氧24h后的TNF-α、IL-1β、IL-6的水平。结果:试验24h后A/R组、Tet组前炎症因子TNF-α[(682.65±32.59)、(388.94±17.22)],IL-1β[189.11±8.29),(109.46±7.62)],IL-6[(78.45±2,74)、(40.91±1.53)]水平(pg/ml)均较对照组的[(211.44±14.20),(59.64±2.43),(23.99±3.24)]显著升高(P<0.01),Tet组前炎症因子TNF-α、IL-1β、IL-6水平均分别显著低于A/R组的(P<0.01)。结论:Tet可以抑制IκB-α磷酸化,显著减少前炎症因子TNF-α、IL-6的产生,减轻心肌缺血/再灌注损伤。

Objective： To investigate the effect of tetrandrine on anoxia/reoxygenation （A/R） induced proinflammatory factors： TNF-α, IL--1β, IL-6 in cultured cardiocytes of neonate rates. Methods： After cardiocytes were success- fully cultured in vitro, it were randomly divided into three groups： control group, A/R group, tetrandrine （Tet） group. Control group not treated with A/R, continuous incubated 26h under normal circumstance. In A/R group, cells were incubated in non-saccharide, non-serum culture medium 2h, which saturate by 95% argon gases; then reoxygenation incubate followed, cells were incubated in normal circumstance 24h. In Tet group the procedure of anoxia/reoxygenation was same to A/R group, but added Tet （30μmol/L） into culture fluid and incubated 60 min before anoxia beginning. TNF-α, IL-1β, IL-6 were detected after reoxygenation 24h. Results： After test 24h the proin- flammatoryfactors TNF-α [ （682.65±32.59）, （388.94±17.22）], IL-1β [189.11±8. 29）, （109.46±7.62）], and IL-6 [ （78.45±2, 74）, （40. 91±1.53）] levels （pg/ml） increased significantly in A/R group, Tet group, respectively compared with those of control group [ （211.44±14.20）, （59. 64±2.43）, （23. 99±3.24） pg/ml], P〈0.01 all. The TNF-α, IL-1β, and IL-6 levels of Tet group were significant lower than those of A/R group （P〈 0.01 all）. Conclusion： Tet can attenuate myocardial ischemia/reperfusion injury. Its machanism may be through inhibition the IκB-α phosphorylation and reduce the harmful cytokine TNF-α and IL-6.