Study On Inhibitive Effect Of Aspirin On Proliferation Of Lung Cancer PC14 Cells And Preparation Of Microspheres Aerosol

PurposeTo investigate the effect of aspirin?Asp?on lung cancer PC14 cells and its possible mechanism.On this basis,the dosage form and the route of administration were studied to prepare Asp chitosan microsphere aerosol.Direct application of the drug to the lungs not only can avoid first-pass effects,but also reduce drug side effects and improve patient compliance.MethodsPC14 cells were treated with 0,4,8 and 16 mmol·L^-1 Asp for 24,48 and 72 h.The cells proliferation was examined by MTT and the apoptosis was detected by flow cytometry.In order to study the relationship between PC14 cells proliferation and c-Jun N-terminal Kinase?JNK?activation,PC14 cells were pretreated with 5?mol·L^-11 SP600125?JNK inhibitor?for 30 min,and then cultured for 24,48 and 72 h.The effect of Aspirin on the activation of JNK was detected by Western blot.The Asp-chitosan microspheres?Asp-Cs-MC?were prepared by emulsion cross-linking method.Based on the single factor test,the average particle size,drug loading and encapsulation efficiency were taken as the indexes.The value OD was used as the evaluation index,and the Central Composite Design Response Surface Methodology was used to optimize the preparation process.The microspheres were characterized by Fourier transform infrared spectroscopy?FT-IR?,scanning electron microscopy?SEM?and particle size distribution.The dosage form of Asp-Cs-MC was designed into a quantitative inhalation aerosol,and its quality was preliminarily evaluated.ResultsThe results of MTT showed that aspirin inhibited the proliferation of PC14cells in lung cancer,at the same time,aspirin could induce G0-G1 arrest in lung cancer cells.Aspirin inhibited PC14 proliferation by activating JNK and inducing its expression.Central Composite Design Response Surface Methodology optimized the microsphere preparation process.The optimal process conditions were:chitosan concentration 2.3%,the herb around 1.3,oil compared water to4:1,emulsifier to oil phase volume ratio 5%,emulsification speed 800 r·min^-1,emulsification time 3 h,cross-linking agent dosage 1.1 mL,cross-linking time 3h.The resulting particle size distribution and the average particle size is 3.29?m,drug loading is 25.23%,encapsulation efficiency is 63.70%.In vitro release tests demonstrated that Asp-Cs-MC had good sustained release effect.The prepared aerosol had no leakage,could be uniformly sprayed,and the drug content conformed to the regulations,and could be absorbed by the lungs.ConclusionsAsp could inhibit the proliferation of PC14 cells and its mechanism may be related to its enhanced activation of JNK,which provided a theoretical basis for the potential application of aspirin in the treatment of lung cancer.The model established by the Central Composite Design Response Surface Methodology had good predictability.An Asp microsphere type aerosol for pulmonary inhalation administration was prepared.