肝外胆管癌内镜下射频消融术后细胞免疫功能变化的研究

Study on the changes of cellular immune function after endoscopic radiofrequency ablation in extrahepatic cholangiocarcinoma

目的探讨内镜逆行胰胆管造影下射频消融治疗肝外胆管癌对患者细胞免疫功能的影响。方法选取山西省人民医院2019年10月至2022年1月就诊的不可手术切除的78例肝外胆管癌患者进行前瞻性队列研究, 患者均行内镜下射频消融术。取患者治疗前1天、治疗后1周、治疗后1个月外周静脉血, 采用流式细胞术检测淋巴细胞亚群, 采用酶联免疫吸附试验检测细胞因子水平。选取与患者年龄、性别相同的健康体检者78名作为健康对照, 并行相同指标检测。结果患者治疗前1天外周血总胆红素为(193±83)μmol/L, 治疗后1周降至(55±49)μmol/L(t=16.70, P<0.001);治疗前1天γ-谷氨酰转移酶为(242±108)U/L, 治疗后1周降至(92±70)U/L(t=19.12, P<0.001)。患者治疗前1天自然杀伤(NK)细胞、CD3+ T细胞、CD4+ T细胞比例和CD4+ T细胞比例与CD8+ T细胞比例比值(CD4/CD8)、干扰素γ(IFN-γ)水平均低于健康对照组(均P<0.05), CD8+ T细胞比例、调节性T细胞比例均高于健康对照组(均P<0.001), 患者治疗前1天与健康对照组间白细胞介素4(IL-4)水平差异无统计意义(P=0.545)。治疗后1周及1个月患者NK细胞、CD3+ T细胞、CD4+ T细胞比例和CD4/CD8、IFN-γ水平均较治疗前1天升高(均P<0.001), 治疗后1个月均较治疗后1周下降(均P<0.001);治疗后1周及治疗后1个月患者CD8+ T细胞比例、IL-4水平均较治疗前1天下降(均P<0.001), 治疗后1个月均较治疗后1周上升(均P<0.001);治疗后1周和1个月调节性T细胞比例均较治疗前1天下降(均P<0.001), 治疗后1个月较治疗后1周下降(P<0.001)。结论肝外胆管癌患者免疫功能低下。内镜下射频消融术治疗可减轻患者的免疫抑制, 提高细胞免疫水平。

Objective To investigate the effect of radiofrequency ablation treatment of extrahepatic cholangiocarcinoma under endoscopic retrograde cholangiopancreatography on the cellular immune function of patients with extrahepatic cholangiocarcinoma.Methods A prospective cohort study was conducted on 78 patients with inoperable extrahepatic cholangiocarcinoma who visited Shanxi Provincial People's Hospital from October 2019 to January 2022.All of 78 patients underwent endoscopic radiofrequency ablation.Peripheral venous blood was collected from all patients at 1 day before treatment,1 week after treatment and 1 month after treatment.Lymphocyte subsets were detected by flow cytometry,and levels of cytokines were detected by enzyme-linked immunosorbent assay.Seventy-eight healthy subjects with the same age and gender as the patients were selected as healthy controls,and the same indicators were tested.Results The patient's peripheral blood total bilirubin was(193±83)μmol/L at 1 day before treatment,and decreased to(55±49)μmol/L at 1 week after treatment(t=16.70,P<0.001);γ-glutamyltransferase was(242±108)U/L at 1 day before treatment,and decreased to(92±70)U/L at 1 week after treatment(t=19.12,P<0.001).The proportion of natural killer(NK)cells,CD3+T cells,CD4+T cells,ratio of CD4+T cells to CD8+T cells(CD4/CD8)and level of interferonγ(IFN-γ)of patients at 1 day before treatment were lower than those of healthy control group(all P<0.05),the proportion of CD8+T cells and regulatory T cells was higher than those of healthy control group(both P<0.001),and there was no significant difference in the levels of interleukin 4(IL-4)between the patients at 1 day before treatment and the healthy control group(P=0.545).At 1 week and 1 month after treatment,the proportion of NK cells,CD3+T cells,CD4+T cells,CD4/CD8 and IFN-γlevel in patients increased compared with 1 day before treatment(all P<0.001),and decreased at 1 month after treatment compared with 1 week after treatment(all P<0.001).At 1 week and 1 month after treatment,the proportion of CD8+T cells and IL-4 level in patients decreased compared with 1 day before treatment(both P<0.001),and increased at 1 month after treatment compared with 1 week after treatment(both P<0.001).At 1 week and 1 month after treatment,the proportion of regulatory T cells decreased compared with 1 day before treatment(both P<0.001),and decreased at 1 month after treatment compared with 1 week after treatment(P<0.001).Conclusions Patients with extrahepatic cholangiocarcinoma are at a low level of immunity.Endoscopic radiofrequency ablation treatment can alleviate the patient's immunosuppression,enhance the patient's cellular immunity.