Preparation of Tilianin-loaded Nano-micelles and Evaluation of Prevention of Apoptosis in H9c2 Cells / 中国药学杂志

ABSTRACT

OBJECTIVE: To prepare nano-micelles with amphiphilic self-assembly poly (ethylene glycol)-co-poly (propylene sulfide) (PEG-PPS) copolymer as carrier to study the release characteristics of tilianin and investigate its activity to against H9c2 cell apoptosis in vitro. METHODS: An amphiphilic diblock PEG-PPS polymer was used as a carrier material to prepare the tilianin-containing nano-micelles by solvent evaporation. The morphology, particle size and distribution, drug loading and encapsulation rate and in vitro drug release behavior were characterized, H9c2 rat myocardial cell injury model was established by hypoxia/reoxygenation process. Using propranolol (Pro) as a positive control, the morphology of injured cardiomyocytes was observed by microscope. Cell proliferation and cell apoptosis was detected to evaluate the protective effect of blank micelles, tilianin and tilianin loaded nano-micelles on H9c2 cells induced by hypoxia/reoxygenation. RESULTS: Tilianin-loaded nano-micelles was spherical with uniform particle size distribution. The drug loading was 3.82%. The average particle diameter of tilianin-loaded nano-micelles was 137 nm, polydispersity coefficient was 0.162 and the encapsulation efficiency was 91.45%. In vitro drug release studies showed that there was no drug-induced burst release of tilianin-containing nano-micelles and sustained release characteristics, and the presence of hydrogen peroxide significantly promoted the release of tilianin from the nano-micelles. In vitro cytotoxicity experiments showed that when the concentration of tilianin 5 μg•mL-1, the cell viability of tilianin-loaded nano-micelles was significantly higher than the corresponding concentration of tilianin and PEG-PPS polymer nano-micelles. In vitro anti-apoptotic activity experiments show that tilianin-loaded nano-micelles on H9c2 cell apoptosis induced by hypoxia-reoxygenation have a significant inhibitory effect and was provided inhibition of apoptosis with propranolol. CONCLUSION: Tilianin-loaded nano-micelles have uniform particle size and distribution, sustained release and oxidation characteristics, has a significant protective and apoptosis-inhibiting effect on H9c2 cell injury induced by hypoxia-reoxygenation, which can be used as a promising drug delivery system for the treatment of myocardial ischemia-reperfusion injury.