聚多巴胺并木犀草素对细胞毒性淋巴细胞肿瘤杀伤作用的影响  

EFFECTS OF POLYDOPAMINE COMBINED WITH LUTEOLIN ON TUMOR KILLING BY CYTOTOXIC LYMPHOCYTES

作　　者：李程琳 王梓聿 曹丁元 国超凡 王爽[1] 李玲[1] LI Chenglin;WANG Ziyu;CAO Dingyuan;GUO Chaofan;WANG Shuang;LI Ling(School of Basic Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院,山东青岛266071

出　　处：《精准医学杂志》2023年第3期199-204,共6页Journal of Precision Medicine

基　　金：国家自然科学基金项目(82001754)。

摘　　要：目的 探讨聚多巴胺(PDA)并木犀草素(LUT)通过光热治疗对细胞毒性淋巴细胞(CTL)肿瘤杀伤作用的影响。方法 通过扫描电子显微镜(SEM)和接触角测定仪观察和测定合成PDA的表征。将RAW264.7细胞分为Control组、LPS组(2μg/L)、PDA′组(50 000μg/L),常规培养48 h,采用CCK8实验检测细胞活力,通过流式细胞术检测LUT对RAW264.7细胞分化的影响。将雌性C57BL/6小鼠随机分为模型组、PDA组、LUT组以及PDA+LUT组,各组小鼠的后背部均皮下注射B16F10黑色素瘤细胞,建模后第10天在PDA组和PDA+LUT组建模小鼠肿瘤处注射PDA行光热治疗1次后,对建模小鼠右大腿肌肉分别注射PBS、PDA(2.5μg/只)、LUT(500μg/只)、PDA(2.5μg/只)+LUT(500μg/只),观察建模小鼠肿瘤生长以及存活情况。在治疗后第3、7天,取各组建模小鼠脾脏制备单细胞悬液,采用流式细胞术检测小鼠体内巨噬细胞分化和T细胞表达情况。结果SEM观察到合成的PDA具有较强的吸附能力和亲水性,并且CCK8实验检测结果显示PDA对细胞活力无影响(P>0.05)。各组RAW264.7细胞的CD206和iNOS平均荧光强度差异有显著性(F=30.72、1 516.00,P<0.05)。注射或不注射PDA时,则注射或不注射LUT小鼠的肿瘤大小和体内免疫细胞iNOS、CD206、IFN-γ^(+)CD4^(+)、TNF-α^(+)CD8^(+)表达水平比较差异均有显著性(F=23.10~235.52,P<0.05);注射或不注射LUT时,则注射或不注射PDA小鼠的肿瘤大小和体内免疫细胞iNOS、CD206、CD4^(+)IFN-γ^(+)、CD8^(+)TNF-α^(+)表达水平比较差异均有显著性(F=8.98~200.67,P<0.05);建模小鼠中PDA+LUT组存活率与其他组相比显著提高(χ^(2)=9.70,P<0.01)。结论 LUT具有抑制小鼠巨噬细胞向M2分化并促进其向M1分化的作用,PDA并LUT治疗可以有效抑制小鼠肿瘤生长,提高生存率,增强CTL的肿瘤杀伤作用。Objective To explore the effects of polydopamine(PDA)plus luteolin(LUT)with photothermal therapy on tumor killing by cytotoxic lymphocytes(CTL).Methods The synthesized PDA was characterized by using a scanning electron microscope(SEM)and a contact angle analyzer.RAW264.7 cells were divided into control group,LPS group(2μg/L),and PDA′group(50000μg/L).After 48 h culture,cell viability was determined by cell counting kit-8.The effect of LUT on the differentiation of RAW264.7 cells was determined by flow cytometry.Female C57BL/6 mice were randomly divided into model group,PDA group,LUT group,and PDA+LUT group.B16F10 melanoma cells were subcutaneously injected into the back of mice in each group.On the 10th day after modeling,the PDA and PDA+LUT groups received injection of PDA into the tumor as well as photothermal therapy.The mice in the four groups received injection of PBS,PDA(2.5μg/piece),LUT(500μg/piece),and PDA(2.5μg/piece)plus LUT(500μg/piece)into the muscle of the right thigh,respectively.We observed the tumor growth and survival of the model mice.On days 3 and 7 after treatment,the spleen of each mouse was taken to prepare a single cell suspension for analysis of macrophage differentiation and T cell expression by flow cytometry.Results SEM showed that the synthesized PDA had good adhesion and affinity for water.CCK8 showed that the PDA did not affect cell viability(P>0.05).Significant differences were found in the mean fluorescence intensities of CD206 and iNOS between different groups of RAW264.7 cells(F=30.72,1516.00,P<0.05).There were significant differences in tumor size and the expression levels of iNOS,CD206,IFN-γ^(+)CD4^(+),and TNF-α^(+)CD8^(+)of immune cells between mice with and without LUT injection,under the condition of either injecting or not injecting PDA(F=23.10-235.52,P<0.05).There were significant differences in tumor size and the expression levels of iNOS,CD206,IFN-γ^(+)CD4^(+),and TNF-α^(+)CD8^(+)of immune cells between mice with and without PDA injection,under the condition o

关 键 词：黑色素瘤 细胞系 肿瘤 光热疗法 多巴胺 聚合物 木犀草素 肿瘤逃逸 T淋巴细胞 细胞毒性 

分 类 号：R392.5[医药卫生—免疫学] R739.5[医药卫生—基础医学]