| Objective(1) To study the anti-tumor effects of Oxytropis kansuensis Bunge alkaloid fraction on murine ascites sarcoma cell line S180 and research its effect on immunity function at the same time.(2) To investigate the effects on proliferation,cell cycle,cell morphous,adhesion, invasion and motility of Oxytropis kansuensis Bunge alkaloid fraction on human gastric cancer cell line BGC-823 in vitro.Methods(1) Healthy Kunming mice were divided into two groups randomly,named Oxytropis kansuensis Bunge alkaloid fraction group and control group,ten mice in each group. The mice were given alkaloid fraction for 3000 mg·kg-1,the control group was given the same volume of normal saline.Two weeks after the mice were sacrificed,dissected and observed changes in major organs.(2)All of the mice were divided into five groups randomly after they were innoculated of S180 cell line,then they were given normal saline or diffierent doses of Oxytropis kansuensis Bunge alkaloid fraction via intragastric administration for ten days,the 5-FU positive control group were intraperitoneal injection.On the eleventh day all mice were sacrificed and weighed,calculated the tumor inhibitory rate,spleen index and thymus index after they were dissected and determined the spleen cell multiplication by MTT colorimetric assay.(3) Human gastric cancer cell line BGC-823 cultured in vitro were treated with different doses of Oxytropis kansuensis Bunge alkaloid fraction.The effects of alkaloid fraction on the proliferation,cell cycle,apoptosis,cell morphous,adhesion,invasion and motility of human gastric cancer cell line BGC-823 in vitro were explored respectively by MTT colorimetric assay,flow cytometry,inverted microscope and Boyden chamber.Results(1) Acute toxicity experiment:The mice were dissected and observed after they were given high-dose of Oxytropis kansuensis Bunge alkaloid fraction,their body weight and heart,liver,lung,kidney and other important organs had no obvious changes compared with the control group(P＞0.05 ) The two groups of mice were found no obvious abnormalities after the HE staining.(2) By comparison of the tumor inhibitory rate,we saw that Oxytropis kansuensis Bunge alkaloid fraction could inhibit tumor,the effect of the mid-dose was more obviously compared with the negative control group(P＜0.05),but was weaker than the positive control group.(3) Alkaloid fraction could also improve the spleen index and thymus index compared with the two control group(P＜0.05),the spleen cell multiplication had statistics significance (P＜0.05) compared with the two control groups,the effect of the low-dose group was significantly.The spleen cell multiplication on 24h is the highest of the three.(4) MTT colorimetric assay suggested that different concentrations and different times of alkaloid fraction could inhibite the growth of BGC-823 cell line with different degrees and the highest dose of 200μg·mL-1 had the best effect.(5) Flow cytometry to observe the effect to cell cycle of BGC-823 of different doses of Oxytropis kansuensis Bunge alkaloid fraction,indicating the drug effect the cell cycle in a dose-dependent manner,and made the BGC-823 cells arrest in G0/G1 phase,and compared to the control group and the 5-FU group,there was a clear statistical significance(P＜0.01), which the 200μg·mL-1 group on the cell cycle resistance was the most obvious.The drug on the role of apoptosis detection,we found no obvious apoptosis peak.(6) The cell morphological changes were observed with inverted microscope,we found that different doses of the Oxytropis kansuensis Bunge alkaloid fraction could inhibite the growth of BGC-823 cell line.Parts of attached cells shrinked and changed round.At last they floated.Compared with the control group,the cell numbers of the BGC-823 cells were decreased significantly(P＜0.01,P＜0.05).(7) Compared with the control group,the adhesion ability of the BGC-823 cell line was decreased significantly(P＜0.01,P＜0.05) after treatment with Oxytropis kansuensis Bunge alkaloid fraction,the 200μg·mL-1 group on the adherence inhibition ability was the most obvious.Compared with the 5-FU group,the high-dose group is also statistically significant differences in different time periods(P＜0.05).(8) Compared with the control group and the 5-FU group,the invasion abilities of the BGC-823 cell line were decreased significantly(P＜0.01) after treatment with the high-dose of Oxytropis kansuensis Bunge alkaloid fraction(71.67±9.29),the high-dose group was significant differences(P＜0.05) compared with the low-dose group.The invasion abilities of the BGC-823 cell line had no significantly decreased(P＞0.05) after treatment with the low-dose and mid-dose of Oxytropis kansuensis Bunge alkaloid fraction.(9) Compared with the control group and the 5-FU group,the motility abilities of the BGC-823 cell line was decreased significantly(P＜0.01,P＜0.05) after treatment with the high-dose and the mid-dose of Oxytropis kansuensis Bunge alkaloid fraction(78.0±8.19).The motility abilities of the BGC-823 cell line had no significantly decreased(P＞0.05) after treatment with the low-dose of alkaloid fraction.Conclusions(1) Oxytropis kansuensis Bunge alkaloid fraction of high-dose administration on normal mice has no apparent side effects.(2) Oxytropis kansuensis Bunge alkaloid fraction has a certain degree of tumor inhibition on S180 tumor-bearing mice.(3) Oxytropis kansuensis Bunge alkaloid fraction can enhance the immune function on S180 tumor-bearing mice.(4) Oxytropis kansuensis Bunge alkaloid fraction shows a certain inhibitory effect of dose and time-dependent on the BGC-823 gastric cancer cell line in vitro,and can affect the morphology of the tumor cells.(5) The BGC-823 cell cycle is arrested in G0/G1 phase after given Oxytropis kansuensis Bunge alkaloid fraction and S phase cells is reduced,thus the drug affects the cell DNA synthesis and inhibites the cell proliferation.(6) Oxytropis kansuensis Bunge alkaloid fraction can affect adhesion,invasion and motility of the BGC-823 cell line in vitro.
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