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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。
Objective T helper lymphocytes (Th) play an important role in antitumor immunity. In particular, the decrease of Th1 / Th2 ratio is associated with metastasis and recurrence of gastrointestinal malignant tumors. In this study, the effects of different concentrations of ketamine on Th1 and Th2 cells in colorectal cancer patients were investigated in vitro with phorbol myristate-acetate (PMA) and ionomycin. Methods Twenty patients with colorectal cancer and 20 healthy volunteers who were admitted to Shandong Cancer Hospital 2014-10-01-2015-05-31 were enrolled in this study. Peripheral blood mononuclear cells (PBMCs) isolated from each healthy blood sample were added to normal control group (C0 group, n = 20) with equal volume of 0.9% sodium chloride. PBMCs isolated from each blood sample from patients with colorectal cancer were randomly Divided into C1 group, K1 group, K2 group and K3 group, adding 0.9% sodium chloride, 25, 50 and 100μmol / L ketamine (n = 20) respectively. Each group were added PMA and ionomycin 2μL mixed solution incubated 4h after the cells were collected. The proportions of Th1 and Th2 cell subsets were determined by using three-color fluorescent flow cytometry and the ratio (Th1 / Th2) was calculated. Results The proportion of Th1 / Th2 in C0 group was 9.36% in group C and 6.39% in group C1 (P <0.001). The Th1 / Th2 ratio was 2.48 in C0 group and 1.63 in C1 group (P <0.001). Compared with C1 group, there was no significant change in Th1 cells, Th2 cells and Th1 / Th2 ratio in K1 group, P> 0.05; Th1 / Th2 ratio in K2 group increased from 1.63 to 1.71, P <0.001; Th1 cells, Th2 cells decreased, Th1 / Th2 ratio increased, P values were <0.01. With the increase of ketamine concentration, the ratio of Th1 / Th2 increased continuously, P <0.01. Conclusion Colorectal cancer induces immune response and shifts Th1 / Th2 balance to Th2. In vitro PMA and ionomycin stimulation, low-dose ketamine on patients with colorectal Th1, Th2 and Th1 / Th2 ratio had little effect. High concentrations of ketamine can inhibit Th cell differentiation, but increased Th1 / Th2 ratio, and with the increase of concentration, the greater the impact on Th cell differentiation.