The Study Of The Anti-Tumor Activity Of The Anthriscus Sylvestris (L.)Hoffm’ Extracts

Purpose:This study was to investigate the effect of the extracts of Anthriscus sylvestris (L.)Hoffm on inhibiting proliferation of non-small cell lung cancer (NSCLC) A549and H460in vitro, then offer the thesis and experiment proofs for curing NSCLC by the extracts of Anthriscus sylvestris (L.)Hoffm.In vivo, in order to investigate the effect of the extracts of Anthriscus sylvestris (L.)Hoffm on H22in mice, we fully analysis the tumor volume, weight, morphology change and the expression of bax and bcl-2,then offer thesis proofs for curing the tumor by Anthriscus sylvestris (L.)Hoffm.Methods:1.According to the cell model of A549and H460, MTT Assay was applied to detecting the inhibitive effect of different concentration of Anthriscus sylvestris (L.)Hoffm on the proliferation of A549and H460cell lines at different time (24h,48h), then calculate the IC50value of the petrol extract of Anthriscus sylvestris (L.)Hoffm.2.To observe the cell apoptosis character of H460and A549by treating with different doses of the petrol extract of Anthriscus sylvestris (L.)Hoffm.the DAPI method and DNA ladder method were applied.3. To establish the anmimal model by inoculating the H22cell line in the left oxter asepticly, and then devided the mice into6groups randomly, took the CTX group as the positive control, the0.5%CMC-Na as model group, adding the water decoction group, then gave the mice drμgs daily, and weighed once a day.after14days,7mice in each group were killed,took out the tumor,thymus and spleen,and weighed,calculate the mortality,inhibition rate,thymus index,spleen index,and the rate elongation of life.detected the necrosis of tumor and expression of bax and bcl-2by HE and immunohistochemistry.Results:The extracts of chloroform, petroleum, ethyl acetate, butyl alcohol of Anthriscus sylvestris (L.)Hoffm all had the inhibitive effect on the proliferation of A549and H460, except the butyl alcohol extract, the others at least had50%mortality when the drug concentration was about10μg/ml.2.24hours after the A549and H460were treated by the petroleum extract,the IC50value were6.13μg/ml and0.97μg/ml respectively.24hours and48hours later, the lung cancer cells appeared the obviously apopsis phenomenon if treated by the petroleum extract, including the cell collapse, fragmentation of cell nucleus, pyknosis and so on,all of these the most obvious was the fragmentation of cell nucleus,and the apopsis phenomenon would be more and more obvious within48hours,but the cells in the control group were intact.4.the DNA apopsis electrophoresis lines were observed by DNA ladder.5. The the inhibitive rate of high dose group,low dose group,water decoction group and the CTX group were60.80%,68.29%,62.56%,71.52%respectively;the rate elongation of life were21.2%,49.5%,68.8%,40.6%respectively;the thymus index were2.05±0.34,2.24±0.39,2.23±0.41,1.86±0.57respectively,except the CTX group,the others had the obvious difference compare to the model group (p<0.05).the spleen index were4.11±0.55,4.49±0.68,4.47±0.50,3.77±0.61respectively, all groups had the obvious difference compare to the control group (p<0.05).All groups can increase the expression of bax and reduce the expression of bc1-2.Conclusions:The petrol extracts of Anthriscus sylvestris (L)Hoffm had distinct inhibitive effect and inducing apopsis effect on the proliferation of the NSCLC A549and H460.the petrol extract and water decoction can both inhibit the increase of tumor, reduce the weight of tumor, prolong the life of mice, and the effect may be achieved by increasing the expression of bax and reducing the expression of bc1-2.the Anthriscus sylvestris (L.)Hoffm showed the effect on curing tumor and it might be developed as a dr μg for curing tumor.