【摘要】： 目的: 嗜酸性粒細(xì)胞(eosinophil,EOS)由骨髓造血干細(xì)胞增殖分化而來(lái),嗜酸性粒細(xì)胞增高除見(jiàn)寄生蟲(chóng)感染過(guò)敏及皮膚病外,也見(jiàn)于多種腫瘤及血液病。本實(shí)驗(yàn)初步研究嗜酸性粒細(xì)胞抗腫瘤作用,探討其抗腫瘤及其免疫調(diào)節(jié)機(jī)制。 方法: 將人白血病細(xì)胞K562和胃癌細(xì)胞SGC-7901與從致敏小鼠血液和腹水中分離出來(lái)不同濃度的嗜酸性粒細(xì)胞共同培育12h-48小時(shí),應(yīng)用生長(zhǎng)曲線法、MTT比色法來(lái)檢測(cè)EOS對(duì)白血病細(xì)胞K562和胃癌細(xì)胞SGC-7901細(xì)胞株增殖的影響。同時(shí)在小鼠右腋皮下接種肉瘤細(xì)胞(S_(180)),建立荷S_(180)肉瘤小鼠模型,24小時(shí)后隨機(jī)分為4組:空白對(duì)照組、陰性對(duì)照組、陽(yáng)性對(duì)照組、EOS治療組。觀察荷瘤小鼠腫瘤組織生長(zhǎng)情況,處死動(dòng)物分離腫瘤組織,計(jì)算抑瘤率;HE染色觀察瘤組織細(xì)胞形態(tài);MTT法檢測(cè)荷瘤小鼠淋巴細(xì)胞增殖能力;放射免疫法測(cè)定小鼠血清中IL-4、TNF-a的含量。 結(jié)果: 1、血中EOS與胃癌SGC-7901細(xì)胞共培養(yǎng),在48h內(nèi)沒(méi)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(xì)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(dá)19%和22%。 2、EOS對(duì)荷瘤小鼠的腫瘤組織的生長(zhǎng)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(xì)胞增殖反應(yīng)能力明顯增強(qiáng)(P＜0.05),與陰性對(duì)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(jì)學(xué)意義(P＜0.05)。 結(jié)論 1、血液和腹腔液EOS均對(duì)白血病K562細(xì)胞株的增殖有抑制作用,尤以血液EOS抑制K562細(xì)胞株的增殖最強(qiáng)。 2、EOS對(duì)荷S_(180)肉瘤小鼠的腫瘤組織生長(zhǎng)有抑制作用,且促進(jìn)荷瘤鼠T淋巴細(xì)胞增殖及細(xì)胞因子IL-4、TNF-a的產(chǎn)生,提示EOS對(duì)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(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

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