以西罗莫司为基础三联抗肿瘤疗法对大鼠肝癌肝移植复发模型T淋巴细胞的影响

摘要

Objective To explore the influence of triple anti-tumor therapy which bases on sirolimus combined huaier granule and thymosin α-1 on T lymphocyte of rat model with liver cancer recurrence after transplantation.Methods Seventy-two Sprague-Dawley(SD)rats were randomly divided into triple therapy group,sirolimus group,huaier-granule group,thymosin α-1 group,positive-control group and blank group (n=1 2 in each group).Except the blank group,rats in all the other groups were established the simulation animal model of liver cancer recurrence after liver transplantation by chemical-induced method.After the model was established,rats in the positive control group were executed to appraise whether the model was successful.The proportion of regulatory T cells (Treg)of CD4 + T lymphocytes in peripheral blood (Treg%),the percentage of CD4 + T lymphocyte of total lymphocyte(CD4 +T%)and the percentage of CD8 + T lymphocyte of total lymphocyte (CD8 +T%),were detected by the flow cytometry respectively.The relationship between Treg% and CD4 + T %,CD8 + T %,the ratio of CD4 +/CD8 + T lymphocytes(CD4 +/CD8 +)was analyzed by the method of Spearman rank correlation.Results Pathological section of rat liver tissue suggested that the rat model was established successfully.Treg % of positive control group was higher than that of blank group,the difference had statistical significance(P <0.05).Treg% of triple therapy group was significantly lower than that of the positive control group,huaier-granule group,thymosin α-1 group,and significantly higher than the blank group (all in P <0.05 ).Compared with positive-control group,CD4 +T% and CD8 +T% of triple therapy group,sirolimus group and thymosin α-1 group were significantly higher (all in P <0.05).CD4 +T%and CD8 +T% of triple therapy group were significantly higher than those of thymosin α-1 group,sirolimus group and huaier-granule group(all in P <0.05).The relationship between Treg% and CD4 +T%,CD8 +T%, CD4 +/CD8 + in peripheral blood were negatively correlated for rats in each group.In addition,the triple anti-tumor therapy decreased the negative correlation between Treg% and CD4 +/CD8 +.Conclusions Sirolimus based triple anti-tumor therapy can decrease the peripheral blood Treg level of the liver cancer rat,increase the number of T lymphocyte and CD4 +/CD8 +,and play the role of anti tumor cell growth and proliferation.%目的：探讨以西罗莫司为基础联合槐耳颗粒、胸腺肽α-1的三联抗肿瘤疗法对大鼠肝癌肝移植复发模型 T 淋巴细胞的影响。方法72只 Sprague-Dawley （SD）大鼠以随机数字法分为三联组、西罗莫司组、槐耳组、胸腺肽组、阳性对照组、空白组，每组12只。除空白组外，其余各组均采用化学诱癌法建立模拟肝癌肝移植术后复发的动物模型。模型建立后，取阳性对照组大鼠鉴定模型是否成功建立。采用流式细胞术分别检测各组大鼠外周血调节性 T 细胞（Treg）占 CD4＋T 淋巴细胞比例（Treg％）、CD4＋T 淋巴细胞占淋巴细胞总数比例（CD4＋T％）及 CD8＋T 淋巴细胞占淋巴细胞总数比例（CD8＋T％）。采用 Spearman 秩相关分析 Treg％与 CD4＋T％、CD8＋T％及 CD4＋/CD8＋T 淋巴细胞比值（CD4＋/CD8＋）之间的关系。结果大鼠肝癌组织病理切片提示建模成功。阳性对照组的 Treg％高于空白组，差异有统计意义（P ＜0.05）。三联组的 Treg％明显低于阳性对照组、胸腺肽组和槐耳组，明显高于空白组（均为 P ＜0.05）。与阳性对照组比较，三联组、西罗莫司组和胸腺肽组的 CD4＋T％和 CD8＋T％较高，差异有统计学意义（均为 P ＜0.05）。三联组的 CD4＋T％和 CD8＋T％均高于胸腺肽组、西罗莫司组和槐耳组，差异有统计学意义（均为 P ＜0.05）。各组大鼠的外周血Treg％与 CD4＋ T％、CD8＋ T％和 CD4＋/CD8＋均呈负相关，且三联抗肿瘤疗法可降低 Treg％与CD4＋/CD8＋之间的负性相关关系。结论西罗莫司为基础的三联抗肿瘤疗法可降低大鼠外周血 Treg水平，提高 T 淋巴细胞数量及 CD4＋/CD8＋，发挥抗肿瘤细胞生长和增殖的作用。