| Objective:The aim of the study is to investigate the effects of Artemether on the proliferation of C6 glioma cells,to find out that Artemether has inhibitive effect on orthotopic C6 intracranial glioma growth and angiogenesis in tumor bearing rats,and also to explore the possible inhibitive mechanism of Artemether strengthened by ferrous ion.Method:1.MTF(Methy thiazolyl tetrazolium)assay was used to evaluate the inhibitory effect of Artemether treatment on C6 glioma cells.The expression of VEGF in C6 cells treated with Artemether was detected by enzyme-linked immunosorbent assay(ELISA).2.C6 glioma model was constructed by the stereotaxic apparatus.C6 glioma cells were injected to the right caudate nucleus of the rats.Tumor formation was observed and perfusion was done through left ventricular of rat in vivo.The brain was taken and the tumor was observed.HE staining was done to prove that the glioma model was constructed successfully.3.C6 glioma model was constructed by the stereotactic apparatus.Forty eight SD rats were divided into six groups.They were negative control normal saline group(0.4ml /20g/d)as oral;low dose group(33.3mg/kg/d)as oral;middle dose group (50mg/kg/d)as oral;high dose group(66.6mg/kg/d);middle dose plus ferrous sulfate(50mg/kg/d+1.5mg/kg/d)as oral;positive control group Semustine(20mg/ kg/d)ip.The medicine was given for fourteen days and then was stopped in the next six days,total five times like this.Survival time of rats in each group was observed. The survival curve was drown and the survival time of rats in each group was compared by statistical analysis;Stable and reliable brain glioma model in rats was established and SD rats were divided into six groups as the second step.Treatment started on the day when C6 glioma cells were injected and continued until day 10 and the rats had been died at the last time after 24 hours.Inhibitive rates of Artemether were calculated.Tumors were collected for HE staining and growth status of tumor was compared among each group.The expression of vascular endothelial growth factor(VEGF)in tumor cells in each group was observed by immunohistochemistry and was compared by statistical analysis.Results:1.Artemether could inhibit the proliferation of C6 glioma cells and the inhibition depended on the dose and exposure time.The IC50value was 42.26±0.91μg /ml,36.64±0.76μg/ml,and 32.57±0.71μg/ml after treated with Artemether for 24h,48h,and 72h respectively,the concentration(pg/ml)of VEGF in liquid treated with artemether for 48 hours of Artemether group and control group was 148.32±10.72,365.79±9.46,the difference is obvious(P＜0.05).2.Intracranial growth occurred in all implanted rats.3.Survival time of therapy groups and positive control group was obviously longer than the negative control group (P＜0.05);Inhibitive rates of Artemether by oral at dosages of low,middle,high, middle plus ferrous sulfate and Semustine group were 54.5%,61.0%,64.5%,69.8% and 91.2%respectively.The mean of positive expression rates of vascular endothelial growth factor(VEGF)in tumor cells in the negative control normal saline,low,middle,high,middle plus ferrous sulfate and Semustine group were 71.18%,56.80%,52.85%,45.10%,42.33%,37.18%.The expression rate of the negative control group is significant different with every experimental groups (P＜0.05).Conclusions:1.Artemether could inhibit the proliferation of C6 glioma cells and the inhibition depended on the dose and exposure time.Artemether could effectively down regulate the VEGF expression in C6 cells.2.Artemether could prolong the median survival time of intracranial glioma in rats.3.Artemether may suppress the glioma tumor growth probably by inhibiting Angiogenesis.4.Ferrous ion could strengthen the inhibitive effect of the Artemether. |
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