目的 筛选细胞穿膜肽（transcription activator，TAT）和聚乙二醇（polyethylene glycols，PEG）双修饰田蓟苷复合磷脂脂质体（TAT & PEG tilianin CPL，T&PTCPL）的最佳制备工艺，并考察其对心肌细胞H9C2的保护作用。方法 采用薄膜分散-超声法制备T&PTCPL，利用3因素3水平的Box-Behnken设计，分别以总磷脂与田蓟苷质量比（X₁）、DSPE- PEG₂₀₀₀-TAT的浓度（X₂）和水化体积（X₃）为考察对象，包封率（Y₁）、粒径（Y₂）和多分散系数（PDI，Y₃）为主要评价指标筛选T&PTCPL最佳配方，并以Na₂S₂O₄建立H9C2细胞缺氧/复氧损伤模型，以超氧化物歧化酶（SOD）、肌酸激酶同工酶（CK-MB）、丙二醛（MDA）和乳酸脱氢酶（LDH）为指标，考察T&PTCPL对细胞缺氧/复氧损伤的影响，同时，考察其体外释放率（动态透析法）及人结肠癌Caco-2细胞对田蓟苷原料药和T&PTCPL的吸收情况。结果 T&PTCPL的最佳组合为X₁=20，X₂=1.7%，X₃=3.2 mL；包封率为（86.62±2.51）%，粒径为（149.7±8.2）nm，PDI为0.15±0.05。在体外细胞实验中，与模型组比较，原料药组及T&PTCPL组能显著提高SOD活性，减少MDA的含量及抑制LDH和CK-MB的释放（P<0.05），并且T&PTCPL的效果优于原料药。同时，T&PTCPL在48 h基本释放完全，累积释放率88.65%，Caco-2细胞对T&PTCPL的吸收效果较佳。结论 Box-Behnken实验设计法用于T&PTCPL的优化筛选是可行的，数学模型的预测值与实验观察值相符，并且T&PTCPL在体外释放中表现出较好的缓释效果，能促进田蓟苷在Caco-2细胞中的吸收，同时，提示T&PTCPL具有保护心肌缺血再灌注损伤作用。

Objective To optimize the preparation technology of transcription activator (TAT) and polyethylene glycols (PEG) co-modified tilianin-loaded composite phospholipid liposome (TAT & PEG tilianin CPL, T&PTCPL) and investigate its protective effect on cardiomyocytes. Methods The composite phospholipid liposome was prepared by thin film-ultrasonic method. A three- factor, three-level Box-Behnken experimental design was employed. The weight ratio of total phospholipid to tilianin (X₁), the concentration of DSPE-PEG₂₀₀₀-TAT (X₂), and hydration volume (X₃) were observed. The encapsulation efficiency (Y₁), particle size (Y₂), and polydispersion coefficient (Y₃) were evaluated to optimize optimal formula. In addition, hypoxia/reoxygenation model was established with Na₂S₂O₄ in H9C2 cells. Superoxide dismutase (SOD) activity, malonaldehyde (MDA) level and release of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) were assessed to evaluate the effect of T&PTCPL, meanwhile, the in vitro release rate (dynamic dialysis method) and absorption rate of tilianin and T&PTCPL in Caco-2 cell were examined. Results The optimal formula was as following:X₁=20, X₂=1.7%, and X₃=3.2 mL; The encapsulation efficiency was (86.62 ±2.51)%, particle size was (149.7 ±8.2) nm and PDI was 0.15 ±0.05. Compared with model group, T&PTCPL and tilianin groups increased SOD activity, inhibited level of MDA, LDH and CK-MB leakage (P < 0.05), and the effect of T&PTCPL group was better than tilianin group, meanwhile, T&PTCPL was completely released at 48 h, with a cumulative release of 88.65%, and Caco-2 cells had better absorption of T&PTCPL. Conclusion The Box-Behnken design is suitable for optimizing the formulation of T&PTCPL, and the observed responses are in close agreement with the predicted values of the mathematic models; Moreover, T&PTCPL shows a better sustained release effect in vitro release, which promots the absorption of tilianin in Caco-2 cells and suggests that T&PTCPL may have protective effect on myocardial ischemia reperfusion injury.

国家自然科学基金资助项目（H0203）；新疆维吾尔自治区自然科学基金青年基金项目（2015211B029）