金纳米星/胶原复合基质材料对新生大鼠心室肌细胞氧化应激损伤的影响

Effect of Au NanoStars/collagen matrix material on oxidative stress damage in ventricular myocytes of newborn rats

目的通过体外研究验证金纳米星/胶原(Au nanostars/collagen,Au NSs/Col)复合材料具有抗新生大鼠心室肌细胞氧化应激损伤的作用。方法 (1)构建不同浓度Au NSs/Col复合基质材料,利用Live/dead细胞染色、细胞增殖实验(CCK-8)筛选材料的最佳浓度,和Col共用于后续实验;(2)新生大鼠心室肌细胞(NRVM)随机分为Col组、Au NSs/Col组、过氧化氢(H2O2)诱导Col组、H2O2诱导Au NSs/Col组。6 h后,采用AnnexinⅤ-FITC/碘化丙啶(PI)/4',6-二脒基-2-苯基吲哚(DAPI)染色观察细胞的凋亡情况,蛋白质印迹法检测凋亡相关蛋白B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关x蛋白(Bax)的表达。结果 (1)Live/dead和CCK-8实验结果均提示,0.1 mg/ml Au NSs/Col复合材料对NRVM无毒性,且能进一步促进细胞增殖;(2)与未诱导组相比,H2O2诱导后的Col组和Au NSs/Col组细胞的早期凋亡率均明显增加,Bcl-2/Bax比值降低,氧化应激损伤模型成功建立;H2O2诱导后,与Col组相比,Au NSs/Col组细胞早期凋亡率降低,Bcl-2/Bax比值升高。结论 Au NSs/Col复合材料对体外培养的心肌细胞氧化损伤具有保护作用。

Objective To verify antioxidation of Au Nano Stars/collagen（ Au NSs/Col） for ventricular myocytes of newborn rats（ NRVMs） by in vitro studies. Methods（ 1） Different concentrations of Au NSs/Col composite materials were created. The optimum concentration of the material was selected by Live/dead staining and Cell Counting Kit-8（ CCK-8）and Col was used for subsequent experiments.（ 2） NRVMs were randomly divided into Col group,Au NSs/Col group,H_2O_2-induced Col group,and H_2O_2-induced Au NSs/Col group. After 6 h treatment,apoptotic cell morphology and early cell apoptosis rate were observed with Annexin Ⅴ-FITC/propidium iodide（ PI）/4＇,6-diamidino-2-phenylindole（ DAPI） and the expressions of apoptosis related proteins-B-cell lymphoma-2（ Bcl-2） and Bcl-2 associated x protein（ Bax） were detected by Western blotting. Results（ 1） Both Live/dead and CCK-8 experiments indicated that the Au NSs/Col composite material with 0. 1 mg/ml was nontoxicity to NRVMs and could further promote their proliferation.（ 2） Compared with the uninduced group,the early apoptosis rate of the Col group and the Au NSs/Col group after H_2O_2 induction was significantly increased,while the Bcl-2/Bax ratio was decreased,indicating that the oxidative stress damage model was established. After H_2O_2 induction,compared with the Col group,the early apoptosis rate of the Au NSs/Col group was decreased,but the Bcl-2/Bax ratio was increased. Conclusion Au NSs/Col composite material has protective effect on the oxidative damage of cardiomyocytes cultured in vitro.