合欢皮总皂苷对H22荷瘤小鼠细胞免疫功能的影响

Effects of Total Saponins From Albiziae Cortex on Cellular Immunity in H22 Tumor-bearing Mice

目的:观察合欢皮总皂苷对H22荷瘤小鼠CD4+和CD8+T淋巴细胞亚群、细胞因子白细胞介素-2(IL-2)和肿瘤坏死因子-α(TNF-α)水平,凋亡诱导因子(Fas),凋亡诱导因子配体(Fas L)和增殖细胞核抗原(PCNA)蛋白表达的影响,探讨其对细胞免疫功能的干预机制。方法:昆明种小鼠60只随机分为正常组、模型组、环磷酰胺组、合欢皮总皂苷高、中、低剂量组,建立小鼠H22荷瘤模型。给药组分别按环磷酰胺20 mg·kg-1·d-1,合欢皮总皂苷4,2,1 mg·kg-1·d-1ip,连续治疗11 d后,检测抑瘤率,血清IL-2,TNF-α水平,免疫组化染色检测肿瘤细胞Fas,Fas L和PCNA蛋白的表达;苏木素-伊红(HE)染色观察肿瘤组织形态。结果:与模型组比较,合欢皮总皂苷高、中、低剂量组小鼠移植瘤的平均质量均显著下降(P<0.05,P<0.01),合欢皮总皂苷中剂量组能明显提高CD4+T细胞,CD8+T细胞数量(P<0.05,P<0.01),对CD4+/CD8+无影响,合欢皮总皂苷中、高剂量组IL-2水平明显上升(P<0.05),TNF-α水平无变化,合欢皮总皂苷各剂量组Fas L,PCNA蛋白表达显著降低(P<0.01),中、高剂量组Fas表达水平显著升高(P<0.01)。结论:合欢皮总皂苷能够调节细胞免疫功能,其机制可能与其上调CD4+,CD8+T细胞亚群,促进IL-2分泌,提高Fas蛋白表达并抑制Fas L,PCNA蛋白表达有关。

Objective： To observe the effects of total saponins from Albiziae Cortex（ SA） on CD4＋and CD8＋T lymphocyte subsets,levels of interleukin（ IL）-2 and tumor necrosis factor（ TNF）-α,apoptosis inducing factor（ Fas）,apoptosis-induced ligand（ Fas L） and proliferating cell nuclear antigen（ PCNA） protein expressions in tumor-bearing mice,and to explore its intervention mechanism of cellular immune functions.Method： Totally 60 Kunming mice were randomly divided into normal group,model group,cyclophosphamide（ CTX） group,SA high,medium and low dose groups. The H22 tumor bearing mice models were established. CTX（ 20 mg·kg- 1·d- 1）,SA（ 4,2,1 mg·kg- 1·d- 1） were given to the mice of experimental groups by intraperitoneal injection respectively. After continuous treatment for 11 days,the tumor inhibition rate,the levels of IL-2 and TNF-α in serum were detected,and the expressions of Fas,Fas L and PCNA protein expressions in tumor cells were determined by immunohistochemical staining; tumor tissue morphology was observed by htoxylineosin（ HE）staining. Result： As compared with the model group,the average weight of transplanted tumor in various dose groups of SA reduced significantly（ P 〈0. 05,P 〈0. 01）. The number of CD4＋and CD8＋T cells was increased obviously in medium dose group（ P 〈0. 05,P 〈0. 01）,but CD4＋/ CD8＋ratio was not affected. The levels of IL-2 in medium and high dose groups were significantly increased（ P 〈0. 05）,and the levels of TNF-α had no change. The Fas L and PCNA protein expressions were significantly decreased in various dose groups of SA（ P 〈0. 01）,and Fas expression level in medium and high dose groups was significantly elevated（ P 〈0. 01）.Conclusion： SA have immunomodulatory effects on cellular immunity. The mechanisms may be associated with upregulating CD4＋and CD8＋T cell subsets,promoting IL-2 secretion,increasing Fas protein expressions and inhibiting Fas L and PCNA protein expressions.