【摘要】： 目的: 嗜酸性粒細胞(eosinophil,EOS)由骨髓造血干細胞增殖分化而來,嗜酸性粒細胞增高除見寄生蟲感染過敏及皮膚病外,也見于多種腫瘤及血液病。本實驗初步研究嗜酸性粒細胞抗腫瘤作用,探討其抗腫瘤及其免疫調(diào)節(jié)機制。 方法: 將人白血病細胞K562和胃癌細胞SGC-7901與從致敏小鼠血液和腹水中分離出來不同濃度的嗜酸性粒細胞共同培育12h-48小時,應用生長曲線法、MTT比色法來檢測EOS對白血病細胞K562和胃癌細胞SGC-7901細胞株增殖的影響。同時在小鼠右腋皮下接種肉瘤細胞(S_(180)),建立荷S_(180)肉瘤小鼠模型,24小時后隨機分為4組:空白對照組、陰性對照組、陽性對照組、EOS治療組。觀察荷瘤小鼠腫瘤組織生長情況,處死動物分離腫瘤組織,計算抑瘤率;HE染色觀察瘤組織細胞形態(tài);MTT法檢測荷瘤小鼠淋巴細胞增殖能力;放射免疫法測定小鼠血清中IL-4、TNF-a的含量。 結果: 1、血中EOS與胃癌SGC-7901細胞共培養(yǎng),在48h內(nèi)沒有表現(xiàn)出明顯抑制作用(P＞0.05)。血液、腹腔液EOS與白血病K562細胞共培養(yǎng),36h后表現(xiàn)出不同程度的抑制作用(P＜0.05),血液和腹腔液EOS抑制率分別可達19%和22%。 2、EOS對荷瘤小鼠的腫瘤組織的生長有抑制作用,抑瘤率為16%。 3、在ConA刺激下,EOS治療組小鼠T淋巴細胞增殖反應能力明顯增強(P＜0.05),與陰性對照組比較,EOS治療組小鼠血清IL-4(0.69±0.32ng/ml)、TNF-a(1.36±0.36ng/ml)水平顯著升高,有統(tǒng)計學意義(P＜0.05)。 結論 1、血液和腹腔液EOS均對白血病K562細胞株的增殖有抑制作用,尤以血液EOS抑制K562細胞株的增殖最強。 2、EOS對荷S_(180)肉瘤小鼠的腫瘤組織生長有抑制作用,且促進荷瘤鼠T淋巴細胞增殖及細胞因子IL-4、TNF-a的產(chǎn)生,提示EOS對荷瘤小鼠的免疫功能有正調(diào)節(jié)活性。
[Abstract]:Objective: eosinophilic granulocyte (eosinophil,EOS) is derived from the proliferation and differentiation of bone marrow hematopoietic stem cells. In this study, we studied the anti-tumor effect of eosinophil and its anti-tumor mechanism. Methods: human leukemia cell line K562 and gastric cancer cell SGC-7901 were cultured with different concentrations of eosinophils from sensitized mouse blood and ascites for 12h-48 hours. MTT colorimetric assay was used to detect the effect of EOS on the proliferation of leukemia cell line K562 and gastric cancer cell line SGC-7901. At the same time, mice were inoculated subcutaneously with sarcoma cells (S180),) in the right axilla of mice. After 24 hours, the mice were randomly divided into four groups: blank control group, negative control group, positive control group and EOS treatment group. Tumor tissue growth was observed in tumor-bearing mice, tumor tissue was isolated from animals and tumor inhibition rate was calculated. HE staining was used to observe the morphology of tumor tissue, and lymphocyte proliferation ability was detected by MTT method. The content of IL-4,TNF-a in serum of mice was determined by radioimmunoassay. Results: 1. There was no obvious inhibition of EOS in blood and SGC-7901 cells in gastric cancer within 48 hours (P > 0. 05). Blood, peritoneal fluid EOS and leukemic K562 cells were co-cultured. After 36 hours, the inhibition rates of EOS in blood and peritoneal fluid were 19% and 22% respectively (P < 0. 05). (2) EOS could inhibit the growth of tumor tissue in tumor-bearing mice, and the inhibition rate was 16%. 3. Under the stimulation of ConA, the proliferative ability of T lymphocytes in the EOS treatment group was significantly enhanced (P < 0. 05). Compared with the negative control group, the serum IL-4 in the EOS treatment group was (0. 69 鹵0.32ng/ml). The level of TNF-a (1. 36 鹵0.36ng/ml) was significantly increased (P < 0. 05). Conclusion 1. Blood and peritoneal fluid EOS can inhibit the proliferation of leukemia K562 cell line, especially blood EOS can inhibit the proliferation of K562 cell line. 2EOS could inhibit the growth of tumor tissue and promote the proliferation of T lymphocytes and the production of cytokine IL-4,TNF-a in mice bearing S180 sarcoma, suggesting that EOS has a positive regulatory activity on the immune function of mice bearing S180.
【學位授予單位】：蘭州大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R392

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