【摘要】 目的辅助性T淋巴细胞(Th)在抗肿瘤免疫中发挥重要作用,尤其Th1/Th2比值降低与胃肠道恶性肿瘤转移复发有关。本研究拟在体外佛波醇-十四酸酯-乙酸酯(PMA)和离子霉素刺激下,探讨不同浓度的氯胺酮对结直肠癌患者Th1和Th2细胞的影响。方法选取山东省肿瘤医院2014-10-01—2015-05-31收治的20例结直肠癌患者和20名健康体检者的外周静脉血。健康体检者每份血样分离出的外周血单个核细胞(PBMCs)加入等容积0.9%氯化钠为正常对照组(C0组,n=20);结直肠癌患者每份血样分离出的PBMCs随机分为C1组、K1组、K2组和K3组,分别加入0.9%氯化钠、25、50和100μmol/L氯胺酮(n=20)。各组均加入PMA和离子霉素混合溶液2μL孵育4h后收集细胞。采用三色荧光流式细胞技术测定Th1及Th2细胞亚群比例,并计算两者比值(Th1/Th2)。结果 C0组Th1细胞比例为9.36%,C1组为6.39%,差异有统计学意义,P<0.001;C0组Th1/Th2比值为2.48,C1组为1.63,差异有统计学意义,P<0.001。与C1组比较,K1组中Th1细胞、Th2细胞和Th1/Th2比值均无明显变化,均P值>0.05;K2组中Th1/Th2比值由1.63升高至1.71,P<0.001;K3组中Th1细胞、Th2细胞比例降低,Th1/Th2比值升高,P值均<0.01。随着氯胺酮浓度的增加,Th1/Th2比值不断增高,P值均<0.01。结论结直肠癌诱导免疫应答,使Th1/Th2平衡向Th2偏移。在体外PMA和离子霉素的刺激下,小剂量氯胺酮对结直肠患者的Th1、Th2和Th1/Th2比值影响不大。高浓度氯胺酮能抑制Th细胞分化,但增加Th1/Th2比值,且随着浓度的增加,对Th细胞分化的影响越大。更多还原

【Abstract】 OBJECTIVE To observe the changes of T helper cell subsets(Th1cells and Th2cells)in the immune system in colorectal cancer(CRC)patients and to assess the effects of ketamine on Th1 cells and Th2 cells of in vitro.METHODS Twenty patients with GC and 20 healthy controls were selected as research objects from 2014-10-01 to 2015-05-31,in Shandong Cancer Hospital.The normal control group(group C0,n=20)was PBMCs isolated from the healthy controls added isometric 0.9%NaCl.The PBMCs isolated from the CRC patients were assigned randomly to 4groups(groups C1,K1,K2,K3,with 0.9%NaCl,25,50and100μmol/L katamine,respectively,n=20each).The PBMCs were stimulated with phorbol-myristate-acetate(PMA)and ionomycin for 4h.The percentages of Thl cells and Th2 cells were detected by three-color flow cytometry and Th1/Th2 ratio were then calculated.RESULTS Percentage of the Th1 cells was 9.36%in group C0,which was 6.39%in group C1(P<0.001).The ratio of Th1/Th2 was 2.48 in group C0,which was 1.63 in group C1(P<0.001).Th1 cells,Th2cells and Th1/Th2 ratio had no obvious change in group K1(all P>0.05).The ratio of Th1/Th2 was increased to 1.71 in group K2,which was 1.63 in group C1(P<0.001).The populations of Th1 cells and Th2 cells were decreased,but the ratio of Th1/Th2 was increased in group K3(P<0.001).With the increase of concentrations of ketamine,the Th1/Th2 ratio was increased(all P<0.001).CONCLUSION CRC shifts the balance of Th1/Th2 towards Th2by inducing an immunological response.In our in vitro study,a little dose of ketamine has no influence on the differentiation of Th cells,but anesthetic concentrations of ketamine can suppress the Th cell differentiation,increase the Th1/Th2 ratio.Ketamine can affect the differentiation of Th cells in a dose-dependent model.更多还原