黄芩苷PEG-PCL纳米胶束的体外评价、细胞内分布及抗心肌细胞凋亡的研究

Study on in Vitro Evaluation, Intracellular Distribution and Anti-apoptosis of Baicalin PEG-PCL Nanomicelle

关键词:  PEG-PCL纳米胶束  线粒体  黄芩苷  细胞摄取  心肌细胞凋亡

Key words:  PEG-PCL nanomicelle  mitochondria  baicalin  cellular uptake  cardiomyocyte apoptosis

摘要:

目的 评价黄芩苷PEG-PCL纳米胶束的细胞内分布及抗心肌细胞凋亡作用。方法 采用薄膜水化法制备黄芩苷PEG-PCL纳米胶束，以香豆素-6作为荧光探针，评价PEG-PCL纳米胶束在细胞内的摄取及细胞内分布；采用10 μmol·L^-1异丙肾上腺素诱导H9c2心肌细胞凋亡进行造模，将等剂量黄芩苷和黄芩苷PEG-PCL纳米胶束在造模前预处理1 h，测定细胞凋亡相关的caspase 3活性、ROS水平，Bcl-2和Bax蛋白的表达水平。结果 黄芩苷PEG-PCL纳米胶束具有粒径小，释药缓慢等优良特点；荧光试验表明，PEG-PCL纳米胶束能促进药物的细胞摄取，还能将药物聚集在线粒体部位，细胞凋亡试验结果表明，黄芩苷PEG-PCL能显著降低凋亡相关的caspase 3活性、ROS水平，降低促凋亡Bax的表达，明显提高抗凋亡Bcl-2的表达，这些结果均与等剂量的黄芩苷存在显著性差异。结论 黄芩苷PEG-PCL纳米胶束能促进黄芩苷被心肌细胞摄取，有助于将药物聚集在线粒体部位，从而很大程度上提高药物抗心肌细胞凋亡作用。

Abstract:

OBJECTIVE To evaluate the intracellular distribution and anti-cardiomyocyte apoptosis effect of baicalin PEG-PCL nanomicelle. METHODS Baicalin PEG-PCL nanomicelle were prepared by membrane hydration method. Coumarin-6 was used as a fluorescent probe to evaluate the uptake and intracellular distribution of PEG-PCL nanomicelle. Modeling of H9c2 cardiomyocyte apoptosis induced by 10 μmol·L^-1 isoproterenol was performed. The isodose baicalin and baicalin PEG-PCL nanomicelle were pretreated for 1 h before modeling, and the apoptosis-related caspase 3 activity, ROS levels and expression levels of Bcl-2 and Bax proteins were determined. RESULTS Baicalin PEG-PCL nanomicelle had excellent characteristics such as small particle size and slow release rate. Fluorescence tests showed that PEG-PCL nanomicelle can promote the cell uptake of drugs, and can also accumulate drugs in the mitochondria. The results of death test showed that baicalin PEG-PCL nanomicelle can significantly reduce apoptosis-related caspase 3 activity, ROS levels, reduce the expression of pro-apoptotic Bax, and significantly increase the expression of anti-apoptotic Bcl-2. There was a significant difference compared with the baicalin. CONCLUSION Baicalin PEG-PCL nanomicelle can promote the uptake of baicalin by cardiomyocytes and help to accumulate drugs in the mitochondria, thus greatly improving the anti-cardiomyocyte apoptosis effect.