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目的探讨芝类提取物(GLE)对Walker-256腹水大鼠免疫功能的影响及其机制。方法根据不同处理方法将60只Wistar大鼠随机分为6组:对照组、模型组、低剂量GLE组、中剂量GLE组、高剂量GLE组和顺铂组,每组10只。采用Walker-256大鼠腹水癌细胞接种法造模后,高、中、低剂量GLE组大鼠分别给予GLE 0.84、1.68、3.36g/(kg·d)灌胃,每天2次,每次2mL,连续14d;顺铂组大鼠给予0.004g/(kg·d)顺铂(溶于生理盐水)腹腔注射,每周1次。对照组(未造模大鼠)和模型组均给予生理盐水灌胃,每天2次,每次2mL,连续14d。观察各组大鼠的一般情况,测定腹水质量,称量计算大鼠脾、胸腺和肾脏器指数,检测肾功能;流式细胞术检测各组大鼠胸腺和骨髓的细胞周期、外周血T淋巴细胞亚群分布和自然杀伤(NK)细胞数;血清免疫因子蛋白芯片检测对照组、模型组和中剂量GLE组大鼠血清免疫细胞因子的水平。结果 GLE可有效减少大鼠腹水质量(P<0.05)。与对照组相比,模型组和顺铂组均出现胸腺、脾和肾损伤,给予中、高剂量GLE干预后大鼠胸腺指数、肾指数提高,血肌酐和尿素氮值降低(P<0.05);模型组和顺铂组大鼠胸腺和骨髓细胞发生G0/G1期阻滞,给予GLE干预后G0/G1期阻滞减少、G2/M和S期细胞比例升高(P<0.05)。与模型组相比,给予GLE干预使大鼠外周血中CD4+细胞比例、CD4+/CD8+细胞比值和NK细胞比例均增加,CD8+细胞比例降低(P<0.05)。中剂量GLE组大鼠血清中干扰素γ(IFN-γ)、白介素(IL)-1α、IL-1β、IL-2、IL-4、IL-10和肿瘤坏死因子α(TNF-α)的表达与模型组相比均升高(P<0.05)。结论GLE可有效减少Walker-256腹水大鼠腹水生成,改善大鼠免疫功能,且与顺铂相比无肾脏损害。
Objective To investigate the effect and mechanism of GEL on immune function in Walker-256 ascites rats. Methods Sixty Wistar rats were randomly divided into 6 groups according to different treatment methods: control group, model group, low dose GLE group, middle dose GLE group, high dose GLE group and cisplatin group, with 10 rats in each group. After modeling with Walker-256 rat ascites carcinoma cell inoculation method, the GLE rats in high, medium and low dose groups were administered GLE 0.84,1.68,3.36g / (kg · d) orally intragastrically twice daily for 2mL For 14 days. Cisplatin group was given intraperitoneal injection of 0.004 g / (kg · d) cisplatin (dissolved in normal saline) once a week. The control group (non-model rats) and model group were given normal saline gavage, 2 times a day, each 2mL, continuous 14d. The general conditions of rats in each group were observed, the ascites quality was measured, the index of spleen, thymus and kidneys of rats were weighed, the renal function was measured, the cell cycle of thymus and bone marrow of rats in each group were detected by flow cytometry, The distribution of cell subpopulation and the number of natural killer (NK) cells were measured. The levels of serum immune cytokines in control group, model group and middle dose GLE group were detected by serum immunoprotein protein chip. Results GLE can effectively reduce the ascites quality of rats (P <0.05). Compared with the control group, thymus, spleen and kidney damage occurred in model group and cisplatin group. Thymus index and renal index were increased and serum creatinine and urea nitrogen were decreased (P <0.05) G0 / G1 phase arrest occurred in the thymus and myeloid cells in the model and cisplatin groups. The G0 / G1 phase arrest and the G2 / M and S phase cells were increased after GLE treatment (P <0.05). Compared with the model group, the proportion of CD4 + cells, the ratio of CD4 + / CD8 + cells and the proportion of NK cells in peripheral blood were increased and the proportion of CD8 + cells was decreased in GLE group (P <0.05). The levels of IFN-γ, IL-1α, IL-1β, IL-2, IL-4, IL-10 and tumor necrosis factor alpha Compared with the model group, the expression increased (P <0.05). Conclusion GLE can effectively reduce ascites formation in Walker-256 ascites rats, improve immune function in rats, and have no renal damage compared with cisplatin.