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目的:研究透明质酸对小鼠骨髓来源树突状细胞功能的影响以及回输后荷黑色素瘤小鼠脾淋巴细胞增殖、活化和细胞因子的变化,进而探讨透明质酸诱导的树突状细胞增强荷瘤小鼠免疫功能的机制。方法:体外细胞因子联合诱导培养小鼠骨髓细胞获得树突状细胞(DCs),免疫磁珠分选纯化获得CD11c+树突状细胞,经不同浓度透明质酸(HA)刺激后,采用酶联免疫吸附法(ELISA)检测培养上清液中细胞因子IL-12p70含量。建立小鼠皮下B16黑色素瘤模型,肿瘤局部皮下回输HA孵育DC后检测肿瘤大小,应用Con A检测脾淋巴细胞增殖情况,应用MTT法检测脾淋巴细胞杀伤活性,ELISA法检测脾淋巴细胞分泌的TNF-α和IFN-γ的表达,以单纯DC回输、生理盐水注射以及正常小鼠(无瘤)组作为对照。结果:在10~100μg/m L范围内,HA以剂量依赖的方式上调DCs分泌IL-12p70。HA孵育DC处理组肿瘤生长明显受到抑制;淋巴细胞增殖反应、杀伤活性和细胞因子TNF-α和IFN-γ的表达明显高于单纯DC组和生理盐水组(P<0.05)。结论:透明质酸可促进小鼠骨髓DC的成熟;透明质酸孵育的DC通过增强荷瘤小鼠的抗肿瘤免疫功能而抑制肿瘤的生长。
OBJECTIVE: To study the effect of hyaluronic acid on the function of dendritic cells derived from bone marrow of mice and the proliferation, activation and cytokines of spleen lymphocytes in melanoma mice after transfusion, and to explore the effect of hyaluronic acid-induced dendritic cells Mechanism of enhancing immune function in tumor-bearing mice. Methods: Dendritic cells (DCs) were isolated from bone marrow cells of mice induced by cytokines in vitro. CD11c + dendritic cells were isolated and purified by magnetic beads. After stimulated with different concentrations of hyaluronic acid (HA) The content of cytokine IL-12p70 in culture supernatant was detected by ELISA. The mouse subcutaneous B16 melanoma model was established. After the tumor was subcutaneously transfused with HA, DCs were harvested to detect tumor size. The proliferation of splenic lymphocytes was detected by Con A assay. The cytotoxic activity of spleen lymphocytes was detected by MTT assay. The secretion of splenic lymphocytes The expression of TNF-α and IFN-γ were detected by simple DC reinfusion, saline injection and normal mice without tumor. Results: In the range of 10 ~ 100μg / m L, HA up-regulated DCs secretion of IL-12p70 in a dose-dependent manner. The tumor growth of HA-treated DCs was significantly inhibited. The proliferation and cytotoxicity of lymphocytes and the expressions of cytokines TNF-α and IFN-γ were significantly higher than those of DC and saline groups (P <0.05). CONCLUSION: Hyaluronan can promote the maturation of DC in mouse bone marrow. Hyaluronan-incubated DC can inhibit the growth of tumor by enhancing anti-tumor immune function of tumor-bearing mice.