p38MAPK-siRNA脂质体递药系统的制备及体外抗炎作用的初步研究

Preparation and in vitro anti-inflammatory effect study of p38MAPK-siRNA liposome drug delivery system

目的制备一种递送p38MAPK-siRNA的阳离子脂质体,研究其对脂多糖(LPS)诱导的小鼠RAW264.7炎症细胞模型的抗炎作用。方法采用薄膜分散-超声法制备p38MAPK-siRNA阳离子脂质体;用超速离心法分离制剂和游离p38MAPK-siRNA,通过荧光标记法检测脂质体对p38MAPK-siRNA的包封率;选用小鼠RAW264.7单核巨噬细胞为细胞模型,考察制剂的体外抗炎作用。结果成功制备了阳离子脂质体,p38MAPK-siRNA与其的包封率为87.29%;制剂具有一定的体外抗炎作用,通过抑制p38MAPK信号通路的活化,降低相关炎症因子IL-6、TNF-α的表达水平,从而发挥细胞抗炎作用。结论以小分子干扰RNA作为治疗手段的新型递药系统在未来药物治疗领域具有巨大的潜力。

OBJECTIVE To prepare a cation-liposome for p38 MAPK-siRNA delivery, and then to investigate anti-inflammatory effect of the preparation on LPS induced RAW264.7 inflammatory cell model.METHODS The cation p38 MAPK-siRNA liposome was prepared with lipid hydration-sonication method.Free siRNA and siRNA-liposome preparation were separated by ultracentrifugation.Encapsulation efficiency of the preparation was determined by fluorospectrophotomete.Mouse RAW264.7 macrophage cell was selected as a cell model,investigating the anti-inflammatory effect in vitro.RESULTS Cation p38 MAPK-siRNA liposome was successfully prepared with the encapsulation efficiency of 87.29%.The preparation could inhibit inflammation in vitro by inhibiting the activation of p38 MAPK signal pathway,down-regulating the expression of related inflammatory cytokines IL-6 and TNF-α,thus leading to cell inflammation.CONCLUSION siRNA drug delivery system has great potential in the future therapy.