Aim To investigate the permeation mechanism of paclitaxel by enhancers and preparation factors. Methods The fluidity of mucous membrane and membrane protein conformation changes were determined by using electron spin resonance (ESR) when mucous membrane was treated by several enhancers. At the same time，the factors of penetration of lower dissolution drug across the intestinal mucous membrane were studied in three formulas inclusion complex，microemulsion and injection. Results Polyethylene glycol (PEG) 1500，hydroxypropyl-β-cyclodextrin (HP-β-CD) and phospholipid as enhancers could reinforce the permeation of paclitaxle because of loosening of protein conformation in intestinal mucous membrane. Paclitaxel-HP-β-CD inclusion complex and paclitaxel microemulsion as vehicle could significantly increased permeation kinetic rate of paclitaxel with fluid diffuse method. Conclusion Characteristics of enhancing intestinal absorption of poor dissolution drug had been provided with enhancer the change of membrane fluid.