Abstract
This study was to explore the therapeutic effect and mechanism of puerarin (PUE) combined with PEGylated nanoparticles on a rat cerebral infarction cell model. In this context, PEG-PLGA/PUE nanoparticles were prepared by the thin-film hydration method, and the toxicity of PEG-PLGA/PUE nanoparticles to brain capillary endothelial cell (BCEC) was detected by MTT. The BCEC/TF cell model was obtained by induction of BCEC cells with TNF-α. The BCEC/TF cell model was identified by immunofluorescence; the protein expression was detected by western blotting; the expression level of miR-424 in cells was measured by RT-qPCR; the targeting relationship between miR-424 and PDCD4 was confirmed by dual-luciferase reporter assay. We found that PEG-PLGA/PUE nanoparticles prepared by the thin-film hydration method had uniform particle size, regular shape, and good stability and were not toxic to cells. The vWF was widely expressed in the cytoplasm in BCECs. The BCEC/TF cell model was obtained after TNF-α treatment, and tissue factor (TF) was widely expressed on the cell membrane of BCEC/TF cells. Furthermore, it was observed that the PEG-PLGA/PUE nanoparticles showed better therapeutic effect on the BCEC/TF cell model than PUE. PEG-PLGA/PUE nanoparticles and PUE inhibited the expression of PDCD4 protein by increasing the expression of miR-424 in BCEC/TF cells. In summary, the therapeutic effect of PEG-PLGA/PUE nanoparticles on the in vitro cell model of cerebral infarction is better than that of PUE. Moreover, PEG-PLGA/PUE inhibits the expression of PDCD4 protein by lowering the expression level of miR-424 in cells, thereby reducing the hazard of cerebral infarction.
Highlights
Puerarin (PUE), an isoflavone active ingredient extracted from the rhizoma of legumes Pueraria lobata and Kadsura kadsura, has antioxidant, cholesterol-lowering, and blood pressure lowering effects and is commonly used in the treatment of cardiovascular and cerebrovascular diseases [1]
Based on the above advantages, polyethylene glycol (PEG)-polyglycolic acids (PLGA) nanoparticles can prolong the circulation time of the nanodrug loading system in the blood, thereby targeting specific tissues or organs and Advances in Polymer Technology exerting sustained release and prolonged efficacy [2]. It has been documented [3] that PEG-PLGA nanoparticles can achieve deep penetration and have a long residence time in tumor tissues. As both the cerebral infarction site and the tumor tissue have similar inflammatory infiltration, it was speculated that PEG-PLGA nanoparticles could exert similar effects at the cerebral infarction site. erefore, the therapeutic effect and underlying mechanism of PUE combined with PEG-PLGA nanoparticles on a rat cerebral infarction cell model were explored to provide a new theoretical basis for the treatment of cerebral infarction
Determination of Encapsulation Efficiency and Drug Loading. e 500 μL of PEG-PLGA/PUE nanoparticles after micropore filtration was centrifugated at 12000 rpm for 10 min, with high speed and low temperature, and the upper layer was drug-loaded nanoparticles. e encapsulation structure of the nanoparticles was destroyed by adding 10 volumes of methanol solution to completely release PUE, and the drug loading of PUE in the nanoparticles was measured by HPLC and recorded as M-load. e same method was used to calculate the total amount of drug in 500 μL of nanoparticles, recorded as M-total, and the PUE encapsulation efficiency was worked out by using the following formula: PUE encapsulation efficiency M-load/ M-total
Summary
Puerarin (PUE), an isoflavone active ingredient extracted from the rhizoma of legumes Pueraria lobata and Kadsura kadsura, has antioxidant, cholesterol-lowering, and blood pressure lowering effects and is commonly used in the treatment of cardiovascular and cerebrovascular diseases [1]. Based on the above advantages, PEG-PLGA nanoparticles can prolong the circulation time of the nanodrug loading system in the blood, thereby targeting specific tissues or organs and Advances in Polymer Technology exerting sustained release and prolonged efficacy [2]. It has been documented [3] that PEG-PLGA nanoparticles can achieve deep penetration and have a long residence time in tumor tissues. As both the cerebral infarction site and the tumor tissue have similar inflammatory infiltration, it was speculated that PEG-PLGA nanoparticles could exert similar effects at the cerebral infarction site. erefore, the therapeutic effect and underlying mechanism of PUE combined with PEG-PLGA nanoparticles on a rat cerebral infarction cell model were explored to provide a new theoretical basis for the treatment of cerebral infarction
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