| The buckwheat, divided into sweet buckwheat (buckwheat) and bitterness buckwheat is dicotyledonous annual crops. Our country is one of the main producing countries for buckwheat in the world. Sweet buckwheat is rich in the northern part, while the Southwest is full of bitterness buckwheat. The drug value and Nutritional value of buckwheat is paid close attention and its development and utilization become the heat topic gradually. Scientific research both at home and abroad shows that buckwheat is full of nutrients. The pharmacological effects are diverse, such as anti-cancer and anti-tumor, declining blood sugar, blood-fat and blood pressure, anti-oxidation, enhancing the heart, anti-inflammatory, reducing the brittleness of blood vessel, etc. Buckwheat polysaccharide is the active composition extracted from sweet buckwheat flower and leaf, and previous studies showed that it can increase the immune function of the experimental animals. Lots of research shows that polysaccharides have many efficacies, such as anti-tumor, anti-inflammatory, anticoagulant and aging, protect liver, etc. The anti-tumor effect of seaweed polysaccharides, semen armeniacae polysaccharide and encomia total polysaccharides has been proved. However the related research about anti-tumor effect of buckwheat polysaccharide is rare. This experiment mainly discusses the auxiliary therapy effect of buckwheat polysaccharides for S180 sarcomas. Objective This topic establishes models of S180 sarcoma to observe the assistant effect of BP on S180 tumor mice and explore its possible mechanism.Methods1. Establishment of tumor model and subgroup administration Mice solid Tumor model: The abdominal cavity of mice was inoculated S180 cells generated for 6 days. Extracting ascites, and washing 2 times with physiological salt. The armpit of right arm of each mouse was inoculated S180 cell for 2×107 subcutaneously. Subgroup administration according to the following method: BP high ,medium ,and low groups were lavaged BP for 400mg·kg-1·d-1 ,200 mg·kg-1·d-1and 100 mg·kg-1·d-1 respectively; CTX group was performedintraperitoneal injection for 30 mg·kg-1 every other day; BP+CTX group was lavaged BP for 200 mg·kg-1·d-1 and performed intraperitoneal injection for 30mg·kg-1 every other day; Bearing cancer model group was lavaged the same volume saline; The control group was lavaged and performed intraperitoneal injection the same volume saline; Each group was 24 mice and administration for 2 weeks.Mice ascites tumor model: The abdominal cavity of mice was inoculated S180 cells generated for 6 days, and each mouse was inoculated cell for 2×107.2. Observation index(1)Solid tumor mice: The existed quality, existed time and the tumor growth inhibition rate were observed dynamic; 35 d existence rate was calculated for each group mice.(2)Ascites tumor mice: Food-intake, behavior performance, mental state, existed time, and excrement form were observed. The death and existence days were recorded in order to observe the existence rate for 25 days.(3)The immunity function index:①Thymus gland (spleen) index: The thymus gland and spleen were obtained to weigh and calculate thymus gland (spleen) index.②Blood normal regulations: The eyeball was removal to examine blood white cell (WBC), monocyte (MON%), lymphoid cell% (LYM%), neutral grain cell% (Granulocyte %) and hemoglobin (HGB) after administration for 2 weeks.③The lymphoid cell convert function: The proliferation of spleen lymphoid cell was observed on each group with MTT method. (4)The experiment in vitro:①The directly killing effect of BP on S180 tumor cell was observed dynamic with placenta blue dyeing method to calculate cell death percentage.②The cell cycle of tumor cells cultured in vitro was detected by FACS.③The cell apoptosis was detected by PI living cell fluorescence staining method.Results1. Solid tumor mice: The mice of model group and CTX group appeared the decrease of movement and food-intake, angular, dim of skin and muff of reaction and the CTX group was more obvious. The movement and skin color of BP group were better than CTX and control group. BP high, medium and low group could not inhibit the growth of S180 tumor obviously. The 35d existence rate suggested that CTX+BP-M(70.83%), BP-M(62.50%),BP-H(54.17%),BP-L(45.83%),CTX(20.34%),Model(25%) .2. Ascites tumor mice: The appearance and activity of BP group were better than model and CTX groups obviously. BP could improve the existence rate of ascites tumor mice and the 25d existence rate suggested CTX+BP-M(66.67%),BP-M(58.33%),BP-H(50%),BP-L(45.83%),Model(20.83%), CTX(17.81%).3. The result of immunity function examination :(1) compared with the control and CTX group, the thymus gland and spleen index of BP group had no obvious difference. BP-M could inhibit the decrease of the thymus gland and spleen index induced by CTX obviously. (2)The variety of the blood cell composition was improved in BP high ,medium and low group at different degree and, especially, BP-M group could obviously inhibit the decrease of blood cell composition induced by CTX (P<0.01).(3) BP could increase the lymphoid cell conversion function and promote the proliferation of lymphoid cell. Compared with the CTX group, BP-M group could improve the inhibited effect of the lymphoid cell conversion function induced by CTX (P<0.01).4. Experiment in vitro: (1) it was obvious that BP could inhibit tumor cells the proliferation which were cultured in vitro. And BP had inhibiting effect. The most obvious was BP-M group. CTX and CTX+BP-M group could inhibit the proliferation of S180 tumor cells remarkably (P<0.01).(2) S180 tumor cell cultured in vitro detected by FACS on cell cycle, the results showed that the tumor cells in G0 / G1 phase of each dose BP group were increasing, compared with Model group. It showed that BP could stop tumor cells cycle in G0 / G1 phase, inhibited tumor cells from proliferating. Tumor cells could activate their apoptosis mechanisms to apoptosis gradually. Tumor cells in G2 / M period of CTX group increased significantly, compared with Model group (P < 0.01). But Tumor cells in G0 / G1 phase were obviously less than model group (P < 0.05). This showed that the CTX could block the tumor cell proliferation, and the cells were blocked in G2 / M period.(3) This experiment detected apoptosis through PI living cell dyeing. The results showed BP could inhibit the proliferation of S180 tumor cells obviously which were cultured in vitro. The most obvious was BP-M group.CTX and CTX+BP-M group could inhibit the proliferation of S180 tumor cells remarkably (P<0.01).Conclusion:The results suggested that the anti- tumor effect of BP was not obvious, but it could improve existence quality and prolong life span. Experiment in vitro showed BP could inhibit tumor cells'proliferation and promote their apoptosis. The mechanism may be related with adjusting the body immunity function, lowering adverse reaction of chemotherapy and raising chemotherapy effect. |
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