| Objective:(1)to study the anti pulmonary fibrosis effect of TIL in vitro.(2)A HPLC method for the determination of TIL was established.(3)TIL-PLGA-MS was prepared by SPG membrane emulsification.(4)Investigating and characterizing the morphology of TIL-PLGA-MS.(5)Preliminary study on the release of TIL-PLGA-MS in vitro.Methods:(1)the effects of different concentrations of TIL on human embryonic lung fibroblasts(HFL-1)were studied.(2)A HPLC method for the determination of tilianin was established and its methodology was investigated.(3)The tilianin microspheres were prepared by SPG membrane emulsification.On the basis of single factor experiment,the preparation process was optimized by the star point design response surface method,taking the rotational speed,the amount of polyvinyl alcohol and the amount of PLGA as the factors,the particle size,the ball diameter index and the degree of non adhesion of the microspheres as the response values.(4)The particle size and surface morphology of TIL-PLGA-MS microspheres were investigated by SEM and DSC,XRD and FT-IR.(5)To investigate the release of TIL-PLGA-MS in vitro at different time points.Results:(1)when the concentration of tilianin was more than 30.45ug/ml,the inhibition rate of HFL-1 cell proliferation was more than 50%,and the inhibition of HFL-1 cell proliferation was dose-dependent.(2)In the concentration range of 5.16-206.4 ?g/m L,the linear relationship between the content and peak area is good,the accuracy of content determination method is good,the repeatability and recovery rate meet the requirements.(3)The best preparation process of TIL-PLGA-MS is: accurately weigh 0.08 mg of tilianin,80 mg of PLGA,add4 ml of dichloromethane as the organic phase,measure 75 ml of PVA with concentration of 0.83% as the aqueous phase,add the organic phase into the pressure kettle of SPG membrane emulsification instrument,disperse it into 0.83% PVA solution at the speed of920 r/min at the pressure of 0.005 MPa,and stir it for 3 hours at 40 ?;The microspheres were separated by centrifugation,washed with distilled water for three times,thentransferred to the freeze-drying dish,added with 5% mannitol as the freeze-drying protective agent,pre frozen at-80 ? for 12 hours,then transferred to the freeze-drying machine for 12 hours to prepare TIL-PLGA-MS.(4)The TIL-PLGA-MS was spherical or quasi spherical in appearance,similar in size and uniform in dispersion,with an average particle size of(8.22 ± 0.34)?m.The results of FTIR showed that the signal of tilianin and PLGA in TIL-PLGA-MS changed greatly,but the signal of PLGA did not change,which indicated that the formation of TIL-PLGA-MS was the result of the interaction between tilianin and PLGA;the results of X-ray diffraction showed that TIL and PLGA in the microspheres were amorphous,and the interaction between tilianin and PLGA was not only inclusion.The results of DSC showed that there might be interaction between tilianin and PLGA.(5)In vitro release studies showed that the drug was basically released in about6 hours,while TIL-PLGA-MS was still released in 6-10 hours.Conclusion:(1)tilianin can inhibit the proliferation of human embryonic lung fibroblasts.(2)There was a good linear relationship between the two methods.(3)The optimized preparation method is simple and stable.(4)The results of DSC,XRD and FT-IR showed that the target drug was contained in the microspheres.(5)The results of in vitro release showed that the microspheres had a sustained-release effect. |
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