The Study On The Preparation Of Brucea Javanica Oil-based Iodized Oil And Its Cellular Biological Activity

Hepatocellular carcinoma(LC)is recognized as one of the most common malignant tumors in life,with high degree of malignancy,untimely detection,easy metastasis,poor prognosis and high mortality.At present,one of the most commonly used methods for the treatment of liver cancer is transcatheter arterial chemoembolization(TACE),which embolizes the blood supply arteries of liver tumors on one hand,and carries chemical drugs for targeted killing tumor cells on the other.Due to its characteristics of selective aggregation,angiography and anticancer drug carrier,iodinated oil is widely used in TACE.However,iodide oil itself does not have anticancer activity,merely plays an auxiliary therapeutic effect,brucea javanica oil(BJO),an extract from Chinese herbal medicine,has been proved to inhibit a variety of tumor cells.Therefore,in this paper,iodide oil with both embolization and anticancer effects was prepared from BJO,and its anti-tumor biological activity and mechanism were investigated.Firstly,this study explored the synthesis of BJO-based iodide oil and tried to solve the problem of unstable quality and low iodide content(IC)by solvent crystallization to remove ethyl ester of saturated fatty acid and compounded system.The solvent crystallization method for removing saturated fatty acid ethyl ester was as follows,ethanol: ethyl ester 3:1,crystallization temperature-20°C,crystallization time 7 d,freezing crystallization twice,the saturated fatty acid decreased from 12.97% to 4.07%.The optimal iodization process was as follows,phosphoric acid: ethyl ester(mol: mol)= 12:1,potassium iodide: ethyl ester(mol: mol)= 3:1,reaction time 10 h and temperature 70°C,the iodine content was 38.62%.The embolization model in vitro shows that BJO-based iodide oil prepared in this paper has good embolization properties.Secondly,the antitumor bioactivity of BJO and BJO-based iodide oil was evaluated.The results showed that BJO and BJO-based iodide oil effectively suppressed the cell growth and facilitated cell apoptosis of hepatocellular carcinoma cells in a dose-and time-dependent manner.As a results,BJO and BJO-based iodide oil decreased mitochondrial membrane potential and ATP levels,up-regulated the expression levels of C-caspase3,C-caspase9 and Bax and down-regulated the Bcl-2 level.Moreover,the phosphorylation level of AKT and mTOR were decreased,while that of AMPK was increased,and the agonist of PI3K/AKT attenuated the biological effects and related protein expression of BJO on the survival rate and apoptosis of HepG2 cells,which indicated that BJO and BJO-based iodide oil might probably induce HepG2 cell apoptosis by interacting with the PI3K/AKT signal pathways.Finally,the antitumor effect of BJO-based iodide oil combined with oxaliplatin(OXA)was studied.It was found that BJO-based iodide oil combined with OXA can effectively improve the sensitivity of HepG2 cells to OXA.Compared with OXA alone,the addition of BJO-based iodide oil can effectively reduce cell survival rate and increase cell apoptosis rate.In addition,our research also found that BJO-based iodide oil has a certain resistance reversal effect.BJO-based iodide oil(50 ?g/mL)could reverse the drug resistance of HepG2 cell line resistant to OXA(HepG2 / OXA)to a certain degree,and the reversal factor is 2.59 times.