Study On Anti-lung Cancer Effects Of Artemisia Argyi Essential Oil And Artemisia Argyi Essential Oil Niosomes

Artemisia argyi belongs to compositae and is the dry leaves of Chinese mugwort.The essential oil extracted from Artemisia argyi?AAEO?is one of the main active ingredient groups of Artemisia argyi.It has anti-bacterial,anti-inflammatory,antitussive and antiasthmatic effects.AAEO has anti-HBV and anti-hepatocarcinoma activity confirmed by our group's early research.Pre-experimental results showed that AAEO could significantly inhibit the proliferation of lung cancer A549 cells.This topic will study the anti-lung cancer activity of AAEO in vitro and explore its mechanism initially.To prepare AAEO niosomes and evaluate it.The anti-lung cancer activity and pharmacokinetics of AAEO and AAEO niosomes were discussed in this study.The experimental contents include the following four aspects:1.Activity and preliminary mechanism of AAEO against lung cancer A549 cells in vitroThe effect of AAEO on the proliferation of A549 cells was detected by MTT assay.When the AAEO concentration was 660.8?g/mL,the cell growth inhibition rate was 85.75%at 48 hours,and the cell growth inhibition rate at 72hours was 88.37%.The inhibition rate of cell proliferation increased from37.25%to 88.37%at 72 hours with AAEO concentration changed from 188.8?g/mL to 660.8?g/mL.The IC₅₀0 value of 24,48,and 72 hours were 513.54?g/mL,495.60?g/mL,and 484.27?g/mL,respectively.The results showed that AAEO inhibits the proliferation of A549 cells apparently and it was time and concentration-dependent?P<0.05?.The apoptosis of A549 cells was detected by Annerxin V-FITC/PI double staining method.A549 cells were treated with AAEO for 48 hours,the apoptotic rate were 4.8%and 67.1%when AAEO concentration was changed from 188.8?g/mL to 424.8?g/mL,respectively.Which indicated that AAEO caninduceapoptosisofA549cellsinaconcentration-dependent manner?P<0.05?.The cell cycle changes of A549 cells were detected by PI single staining.A549 cells were treated with AAEO for 48 hours,the percentage of cells in G₀/G₁phase decreased from 67.73%to 52.10%,and S phase increased from 26.83%to40.47%with AAEO concentration increased from 0 to 330.4?g/mL.There was no significant change in G₂/M phase.It can be seen that AAEO can arrest cells in S phase.It is speculated that it is related to inhibiting cell DNA synthesis.2.Preparation and evaluation of AAEO niosomesA method was established for determining the multi-component content of AAEO niosomes and methodological was examined.AAEO niosomes were prepared by film ultrasoonic method.The best prescription and process was selected by single factor investigation and Box-Behnken response surface design.The HLB value was 4.88,the AAEO content was 0.141 g,the hydration temperature was 68?,the hydration time was 30 minutes,and the ultrasonic time was 10 minutes.Under these conditions,the encapsulation efficiency was more than 80%.The AAEO niosomes was ivory-white suspension under natural light.The scanning electron microscopy?SEM?result showed that it was uniform spherical with bimolecular structure.The average particle size was407.54±11.37 nm,PDI was 0.114±0.006 and the Zeta potential was-47.0±0.72 mV.The average entrapment efficiency of eucalyptus oil,camphor and borneol were82.32%?82.64%?88.32%,and the average drug loading were 5.26%?1.11%?1.13%,respectively.The results of drug release in vitro showed that the cumulative release of AAEO and AAEO niosomes were 76.18%and 66.07%at12 h,respectively.And the release rate of AAEO niosomes was slower than AAEO.The stability test showed that AAEO niosomes was stable under strong light and high humidity,and it was unstable under high temperature.It was stable when stored at 4?for 90 days.The encapsulation efficiency decreased from 89.25%to 71.47%after 30 days at 25?.At the same time,encapsulation rate dropped to 49.25%in 90 days.It is suggested that AAEO niosomes should be stored at 4?.3.Study on pharmacodynamics of AAEO and AAEO niosomes anti-lung cancer in vivoLung cancer model was established by subcutaneous injection of A549cells.It was randomly divided into 8 groups:negative control group,high,middle and low AAEO groups;high,middle and low AAEO niosomes groups;positive control group?5-Fu?.After 15 days administration,the tumor growth inhibitory effect of 5-Fu group was 71.10%.The inhibition rates of AAEO high dose group and AAEO niosomes high dose group were 55.54%and 58.76%respectively.With the prolongation of treated,toxic and side effects,such as loss of appetite and weight,were observed in 5-Fu group.The weight of mice decreased from 19.33 g to 16.35 g.There was no significant change in body weight of AAEO groups and AAEO niosomes groups.4.Study on pharmacokinetics of AAEO and AAEO niosomes in vivoAAEO and AAEO niosomes were administered intragastrically,and the content of AAEO in AAEO niosomes group was the same as AAEO group.The peak blood concentration of AAEO group reached 10.0913?g/mL at 1.5 hours,while AAEO niosomes group reached at 3 hours,the concentration was 10.9533?g/mL.The half-life of AAEO increased from 3.155 hours to 3.993 hours after encapsulating,and the amount of AAEO in plasma increased by 1.29 times.It indicated that AAEO niosomes had slow release effect and increased the bioavailability of AAEO in vivo.In conclusion,AAEO has obvious anti-lung cancer A549 cells activity in vivo and in vitro.Which can inhibit the proliferation of A549 cells by inhibiting the normal transformation of A549 cell cycle,blocking the cell cycle in S phase and inhibiting the synthesis of DNA.AAEO niosomes can solve these problems of AAEO,such as poor soluble in water and strong volatility.At the same time,it can prolong the circulating time of AAEO in vivo.To provide basis for the development of AAEO new anti-cancer drugs.Innovation points:The anti-lung cancer activity of AAEO was studied for the first time.In order to solve the problems of AAEO,such as poor water solubility and strong volatility,AAEO niosomes were developed for the first time.And it is the first time to investigate the anti-lung cancer activity and pharmacokinetics of AAEO niosomes in vivo.