生物聚合物纳米微粒对人肺癌细胞凋亡的作用

目的制备包覆双氰青蒿素（DHA）的纳米微粒，探讨DHA的生物聚合物纳米微粒的理化性质以及对人肺腺癌A549N胞凋亡作用的影响。方法制备四种纳米微粒：几丁聚糖-海藻酸钠-双氰青蒿素（Chitosan—alginate—DHA）、几丁聚糖-氢氧化钠-双氰青蒿素（Chitosan—NaOH—DHA）、明胶-双氰青蒿素（Gelatin-DHA）和透明质酸-双氰青蒿素（HA—DHA）纳米微粒。采用透射电镜观察纳米微粒的形貌并测量粒径大小，使用高效液相色谱测定纳米微粒包覆DHA的包覆率。MTT细胞活性检测：未包覆DHA的纳米微粒放置浓度及种类的筛选；包覆DHA纳米微粒的细胞活性测试。进行AnnexinV—FITC染色以及PI染色，使用荧光显微镜定性分析HA—DHA纳米微粒对A549细胞凋亡作用的影响。结果TEM结果显示，Chitosan—alginate—DHA、Chitosan-NaOH-DH、Gelatin-DHA和HA—DHA粒径大小为5~40nm。由高效液相色谱测得四种纳米微粒包覆DHA的包覆率分别为18％、21％、13％和35％。根据MTT细胞活性检测结果，筛选放置1OμlHA纳米微粒，HA—DHA纳米微粒能显著抑制A549细胞的活性（P〈0．01）。由荧光染色观察到，相比直接加入DHA，HA—DHA纳米微粒能够增强对A549细胞凋亡的作用。结论HA—DHA纳米微粒对A549细胞凋亡有增强作用。

Objective Four types of bin-polymeric nanoparticles were encapsulated by dihydroartemisinin, followed by the characterization and investigation on the apoptotic effect of DHA encapsulated nanoparticles toward lung cancer cells line A549. Methods Four types of bio-polymeric nanoparticles including chitosan-alginate-DHA, chitosan-NaOH-DHA, gelatin-DHA and HA-DHA were prepared. The obtained nanoparticles were characterized by TEM observation, size distribution as well as DHA encapsulation ratio via HPLC. MTT assay was performed to evaluate cell viability： the optimal type and dose of nanoparticles were determined, and MTT assay was performed to evaluate the apoptotic effect of DHA encapsulated within nanopartieles toward cells line A549. The apoptotic effect of DHA was further analyzed by using fluorescent microscopy. Results TEM images showed that the size distribution of prepared nanoparticles ranged from 5 to 40 nm. HPLC results showed that the encapsulation ratio of chitosan-alginate-DHA, chitosan-NaOH-DHA, Gelatin-DHA and HA-DHA nanoparticles were 18%, 21%, 13% and 35%, respectively. MTT resuks indicated that 10Ix 1 HA dose and HA-DHA nanoparticles significantly inhibited the cell viability of cell line A549 （P〈0.01） . The result of fluorescent microscopy showed that HA-DHA could greatly enhance the apoptotic effect toward cell line A549 compared with DHA alone. Conclusion HA-DHA nanoparticles can enhance the apoptotic effect of DHA by increasing the cellular bioavailability.