载丹参酮ⅡA ROS敏感纳米胶束的制备及体外抗炎作用

Preparation and in vitro Anti-Inflammatory Effect of Tanshinone IIA-Loaded ROS-Sensitive Nanomicelles

本研究针对炎症微环境构建了一种ROS敏感纳米给药系统以解决丹参酮ⅡA的递送难题。首先合成了聚乙二醇-聚硫化丙烯(mPEG-PPS)两亲性载体材料,通过^(1)H-NMR进行结构确认,芘荧光探针法测定临界胶束浓度为0.030 1 mg/mL。采用薄膜分散法制备载丹参酮ⅡA的纳米胶束,根据透射电镜可知该纳米胶束为球状结构,分布均匀,粒径为151.4 nm, PDI为0.196,zeta电位为-8.29 mV,体系相对稳定。在ROS条件下纳米胶束有效释放丹参酮ⅡA,释放速率与H_(2)O_(2)浓度有关。该纳米胶束浓度达到最大时,溶血率仍小于5%,说明其具有良好的生物相容性。MTT结果显示该纳米胶束对LPS诱导的炎症巨噬细胞RAW 264.7增殖具有良好的抑制效果;ELISA试剂盒检测RAW 264.7细胞中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的含量均明显减低。综上结果表明,该纳米胶束具有响应炎症氧化微环境优势,在体外抗炎作用方面发挥出巨大潜力。

In this study,a ROS-sensitive nano-delivery system is constructed to address the delivery challenges of tanshinone IIA for the inflammatory microenvironment.The amphiphilic carrier material of polyethylene glycol-poly(propylene sulfide)(mPEG-PPS)is firstly synthesized,and structure confirmation is obtained by ^(1)H-NMR,and critical micelle concentration of 0.0301 mg/mL is determined by using pyrene fluorescence probe method.The nanomicelles loaded tanshinone IIA are prepared by thin film dispersion method.According to the transmission electron microscopy,the nanomicelles are spherical in structure and uniform in distribution,The particle size is 151.4 nm,the PDI is 0.196,the zeta potential is-8.29 mV,and the system is relatively stable.The nanomicelles effectively release tanshinone IIA under ROS conditions,the release rate is related to H_(2)O_(2)concentration.The hemolysis rate is still less than 5%when the concentration of this nanomicelles reaches the maximum,indicating its good biocompatibility.MTT shows that the nanomicelles has better inhibitory effect on LPS-induced proliferation of inflammatory macrophages RAW264.7.The levels of tumor necrosis factorα(TNF-α)and interleukin 6(IL-6)significantly reduce in RAW264.7 cells by ELISA kit.In summary,the nanomicelles have the advantage of responding to the inflammatory oxidative microenvironment and exert great potential for anti-inflammatory effects in vitro.