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目的:制备新型负载藻蓝蛋白(C-phycocyanin,C-PC)的羧甲基壳聚糖(carboxymethyl chitosan,CMC)纳米微球(nanoparticle,NP),探讨藻蓝蛋白羧甲基壳聚糖纳米微球(C-PC/CMCNP)对人宫颈癌He La细胞生长的影响及其分子机制。方法:采用正交分析方法,以CMC和C-PC的质量比、CMC浓度、Ca Cl2浓度为考察因素,通过检测C-PC/CMCNP的粒径和CMC的包封率,优选制备C-PC/CMCNP的最佳条件,并制备C-PC/CMCNP。CCK-8法检测C-PC、CMC和C-PC/CMCNP对He La细胞增殖的影响,流式细胞术检测He La细胞凋亡情况,Westren blotting检测He La细胞中caspase-3蛋白的表达。结果:CMC与C-PC的质量比为1∶2、CMC质量浓度为1 mg/ml,Ca Cl2质量浓度为1 mg/ml为最佳制备条件,最终制备的C-PC/CMCNP的平均粒径为(118.4±2.07)nm,并具有较高的包封率(63.2%),其缓释12 h的累积缓释率超过60%。C-PC、CMC和C-PC/CMCNP均能抑制He La细胞的增殖,诱导细胞凋亡,并促进caspase-3蛋白的表达,但C-PC/CMCNP的作用效果最为显著。结论:优化制备条件得到具有高效缓释性能的C-PC/CMCNP,其对He La细胞显著的抑制作用可能是通过促进caspase-3蛋白的表达进而诱导肿瘤细胞凋亡来实现的。
OBJECTIVE: To prepare a novel carboxymethyl chitosan (CMC) nanoparticle (NP) loaded with C-phycocyanin (C-PC) and investigate the effects of phycocyanin carboxymethyl chitosan nano Effect of microspheres (C-PC / CMCNP) on the growth of human cervical carcinoma HeLa cells and its molecular mechanism. Methods: C-PC was prepared by orthogonal test, the mass ratio of CMC and C-PC, the concentration of CMC and the concentration of CaCl 2 were investigated. The particle size of C-PC / CMCNP and the encapsulation efficiency of CMC were optimized. / CMCNP and prepare C-PC / CMCNP. The effects of C-PC, CMC and C-PC / CMCNP on the proliferation of HeLa cells were detected by CCK-8 assay. The apoptosis of HeLa cells was detected by flow cytometry. The expression of caspase-3 protein in HeLa cells was detected by Westren blotting. Results: The mass ratio of CMC to C-PC was 1: 2, the concentration of CMC was 1 mg / ml and the concentration of CaCl 2 was 1 mg / ml. The optimum preparation conditions were as follows: the average particle size of the final prepared C-PC / CMCNP Diameter of (118.4 ± 2.07) nm, with higher entrapment efficiency (63.2%), and the cumulative slow release rate of 12 h reached over 60%. C-PC, CMC and C-PC / CMCNP could inhibit the proliferation of HeLa cells, induce apoptosis and promote the expression of caspase-3, but the effect of C-PC / CMCNP was the most significant. CONCLUSION: C-PC / CMCNP with high sustained-release properties can be obtained by optimizing preparation conditions. The significant inhibitory effect of C-PC / CMCNP on He La cells may be through promoting the expression of caspase-3 protein and inducing tumor cell apoptosis.