白花蛇舌草总黄酮对MFC胃癌荷瘤小鼠血清肿瘤标志物和免疫功能的影响

Effects of total Flavonoids of Hedyotis diffusa on serum tumor markers and immune function of MFC gastric cancer bearing mice

目的探讨白花蛇舌草总黄酮(TFHD)对MFC荷瘤小鼠血清肿瘤标志物和免疫功能的影响。方法用MFC细胞构建胃癌荷瘤模型。按照体重将造模成功小鼠随机分5组:模型组、对照组和低、中、高3个剂量实验组,每组10只。另设10只正常小鼠为正常组。实验组灌胃TFHD 200,100,50 mg·kg^(-1),对照组腹腔注射环磷酰胺25 mg·kg^(-1),模型组、空白组灌胃和腹腔注射生理盐水,1次/天,连续给药8 d。用酶联免疫吸附法检测血清中癌胚抗原(CEA)、糖类抗原72-4(CA72-4)、白细胞介素-2(IL-2)和白细胞介素-6(IL-6)含量;用免疫组化法检测肿瘤组织中Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核转录因子-κB(NF-κB)表达水平(单位面积平均光密度值)。结果正常组、模型组、对照组和低、中、高3个剂量实验组的CEA含量分别为(849.64±46.98),(1 076.81±75.87),(874.64±84.78),(1 025.00±68.22),(947.10±41.53)和(893.48±79.22) pg·mL^(-1);这6组CA72-4含量分别是(12.15±0.78),(15.99±0.91),(12.47±1.39),(15.14±1.15),(13.82±0.98)和(12.84±0.77) U·mL^(-1);这6组IL-2含量分别是(290.00±23.01),(224.94±23.06),(202.18±8.19),(237.44±27.48),(259.23±9.84)和(261.31±13.55) pg·mL^(-1);这6组IL-6含量分别是(91.57±7.62),(115.93±6.49),(95.44±6.79),(110.39±4.85),(101.02±8.13)和(98.45±6.17) pg·mL^(-1)。模型组、对照组和低、中、高3个剂量实验组的TLR4表达分别为0.32±0.02,0.24±0.01,0.30±0.03,0.25±0.02和0.21±0.02;这5组MyD88表达分别是0.63±0.02,0.32±0.11,0.54±0.09,0.43±0.08和0.38±0.05;这5组NF-κB表达分别是0.46±0.02,0.26±0.05,0.41±0.04,0.37±0.04和0.30±0.09。上述指标:模型组与正常组比较,差异均有统计学意义(均P <0.01);对照组和高、中2个剂量实验组与模型组比较,除NF-κB中剂量实验组外,差异均有统计学意义(均P <0.01)。结论 TFHD能够降低血清肿瘤标志物并调节小鼠免疫细胞因子,改善机体免疫功能,同时下调TLR4、MyD88、NF-κB的表达起到抗肿瘤作用。

Objective To investigate the effect of total Flavonoids of Hedyotis diffusa(TFHD) on serum tumor markers and immune function of MFC tumor-bearing mice.Methods The tumor-bearing model of gastric cancer established with MFC cells.According to the body weight,the successfully modeled mice were randomly divided into five groups:model group,control group and low,medium and high dose experimental groups,10 mice in each group.In addition,10 normal mice were set as the normal group.The mice in the experimental group were treated with TFHD 200,100 and 50 mg·kg^(-1) by gavage,while those in the control group were given intraperitoneal injection of cyclophosphamide 25 mg·kg^(-1).The mice in the model group and normal group were given with gavage and intraperitoneal injection of normal saline,once a day,for continuous eight days.The serum contents of the carcinoembryonic antigen(CFA),carbohydrate antigen 72-4(CA72-4),interleukin-2(IL-2)and interleukin-6(IL-6) were detected by enzyme linked immunosorbent assay.The expressions(integrated option density/unit area) of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88) and nuclear transcription factor-κB(NF-κB) in the tumor tissue were detected by immunohistochemistry.Results The levels of CEA in normal group,model group,control group and low,medium and high dose experimental groups were(849.64±46.98),(1 076.81±75.87),(874.64±84.78),(1 025.00±68.22),(947.10±41.53) and(893.48±79.22) pg · mL^(-1),respectively;the contents of CA72-4 in the six groups were(12.15±0.78),(15.99±0.91),(12.47±1.39),(15.14±1.15),(13.82±0.98) and(12.84±0.77) U·mL^(-1),respectively;the levels of IL-2 in the six groups were(290.00±23.01),(224.94±23.06),(202.18±8.19),(237.44±27.48),(259.23±9.84) and(261.31±13.55) pg·mL^(-1),respectively;the levels of IL-6 in these six groups were(91.57±7.62),(115.93±6.49),(95.44±6.79),(110.39±4.85),(101.02±8.13) and(98.45±6.17) pg·mL^(-1),respectively;the expression of TLR4 protein in model group,control group and low,medium and high dose experimental groups was 0.32±0.02,0.24±0.01,0.30±0.03,0.25±0.02 and0.21±0.02,respectively;the expression of MyD88 protein in the five groups was 0.63±0.02,0.32±0.11,0.54±0.09,0.43±0.08 and 0.38±0.05,respectively;the expression of NF-κB protein in the five groups was0.46±0.02,0.26±0.05,0.41±0.04,0.37±0.04 and 0.30±0.09,respectively.Compared between model group and normal group,the difference of the indicators were significant(all P <0.01);compared between high and medium dose experimental groups and model group except for the middle dose groups of levels of NF-κB,the difference of the indicators were significant(all P <0.01).Conclusion TFHD can reduce serum tumor markers,regulate immune cytokines in mice,improve immune function,and down-regulate the expressions of TLR4,MyD88 and NF-κB to play an anti-tumor role.