Synergic Effects Of Artemisinin And Resveratrol In Cancer Cells And Preparation Of Compound Niosomes

Artemisinin (ART) is as an effective antimalarial, the main mechanism is by binding to the plasmodiums’high-iron content to produce a chemical reaction, the cell membrane to produce "free radicals", so that the single-cell plasmodium is death. When cancer cells need to divide a lot of iron to replicate DNA, so the cancer cells’iron content is higher than normal cells’, so the artemisinin can selectively kill the cancer cells. Resveratrol (Res) was a polyphenolic compound and and was found it had played a strong anti-tumor effect on three stages of starting, occurring and developing in the tumor. And its main mechanism is inducting cancer cells’apoptosis, inhibiting synthesis of DNA and oxidation of human low-density lipoprotein protein (LDL), but also to inhibit peroxidation of membrane lipid, such as reducing the generation of H2O2. But IC50 of both artemisinin and resveratrolr were very high and need larger clinical dosage. Therefore, we consider the two drugs together is used to enhance the anti-tumor effect by the two drugs together is used to enhance the anti-tumor effect by complementary of anti-tumor mechanism of artemisinin and resveratrol. Due to the low toxicity of the two drugs can greatly reduce toxicity of the general antineoplastic and help improve clinical care. In addition, resveratrol and artemisinin is fat-soluble drugs, poor water solubility, poor oral absorption, no targeting and metabolic instability, so the clinical application is greatly limit. Non-ionic surfactant vesicles (niosomes) is an effective drug delivery structure carrier, with similar cells having the same targeting properties with liposomes, but is more stable in vivo. And it can be changed body-distribution of drug, and has advantages of targeting, prolonged action, reduceing toxicity and increasing drugs’ stability and so on. Therefore, we tried to put the two drugs together as compound vesicles to improve the bioavailability of the drug, to play a better anti-tumor effects.The present study suggested that the combination of ART and Res exhibits the anti-proliferative effect in HeLa and HepG2 cells by inducing apoptosis. In the first, we assessed the activity of ART and Res in a small panel of cancer cell lines. MTT assays demonstrated that ART and Res treatment showed a dose-dependent reduction in cell viability in Hela and HepG2 cells. The IC50 of the combination of ART and Res was lower than the IC50 of the drugs used singly. Thus, we evaluated the drug-drug interaction between ART and Res using the CI method and isobologram methodology. The present study demonstrated that combining ART with Res generated a synergistic anticancer effect in HeLa and HepG2 cells. It is further confirmed by AO staining and Annexin V-FITC/PI assay. The results showed that ART and Res can singly accelerated the rate of cells apoptosis in various different periods. Notably, these data suggested that the induction of apoptosis might be one of the mechanisms of the combination of ART and Res. Furthermore, we investigated the migration of single or combination group in HeLa cells. Combination of ART and Res inhibited the migration significantly than that in single group. DCF fluorescence revealed that ROS levels in combination-treated HeLa cells were much higher than those of the control group and single-drug group, suggesting that combination of ART and Res caused oxidative stress, mitochondrial permeability transition, and then caused apoptosis in HeLa cells.To get a stable artemisinin-resveratrol compound niosomes with high trapping efficiency by film hydration method. The formulation was optimized by orthogonal design, and particle size, encapsulation efficacy and ζ-potential were determined. The encapsulation efficacy of the compound niosomes reached 75.80% and 61.50% for resveratrol and artemisinin. By study in vitro release of the compound niosomes, we found that slow-release rate showed the stability of long-circulating niosomes, which can be expected still be stable in the circulation system. Resveratrol or artemisinin can be slowly released to the tumor location.In conclusion, the present study confirmed that combining ART with Res generated a synergism effect in vitro model of HeLa and HepG2 cells. Combining ART with Res is a hopeful strategy in clinical therapy of solid tumor. Meanwhile, the artemisinin-resveratrol compound niosomes can significantly improve the shortcomings of the drug itself low water solubility, slender pharmacological effects and increase bioavailability and stability of drug. Further investigation will be performed on the molecular mechanism of combining ART with Res in vitro and in vivo.