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  • 赵若琳,殷佩浩,倪振华,孙健,徐可,何裕民.芝类提取物改善Walker-256腹水大鼠免疫功能[J].第二军医大学学报,2017,38(8):1016-1022    [点击复制]
  • ZHAO Ruo-lin,YIN Pei-hao,NI Zhen-hua,SUN Jian,XU Ke,HE Yu-min.Ganoderma lucidum extract improves immune function in Walker-256 ascites rats[J].第二军医大学学报,2017,38(8):1016-1022   [点击复制]
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芝类提取物改善Walker-256腹水大鼠免疫功能
赵若琳1,殷佩浩2,倪振华2,孙健2,徐可2,何裕民1*
0
(1. 上海中医药大学中医基础理论教研室, 上海 201203;
2. 上海中医药大学附属普陀医院中心实验室, 上海 200062
*通信作者)
摘要:
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、3.36 g/(kg·d)灌胃,每天2次,每次2 mL,连续14 d;顺铂组大鼠给予0.004 g/(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2 mL,连续14 d。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。
关键词:  灵芝  腹水  免疫  细胞因子类  T淋巴细胞  天然杀伤细胞
DOI:10.16781/j.0258-879x.2017.08.1016
投稿时间:2017-05-19最后修改时间:2017-08-03
基金项目:国家社会科学基金重点项目(12AZD094),中医学一流学科建设项目.
Ganoderma lucidum extract improves immune function in Walker-256 ascites rats
ZHAO Ruo-lin1,YIN Pei-hao2,NI Zhen-hua2,SUN Jian2,XU Ke2,HE Yu-min1*
(1. Department of Basic Theory of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;
2. Central Laboratory, Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China
*Corresponding author)
Abstract:
Objective To explore the effect of Ganoderma lucidum extract (GLE) on immune function of Walker-256 ascites rats and its mechanisms. Methods Sixty Wistar rats were randomly divided into 6 groups:control group, model group, high-dose GLE group, medium-dose GLE group, low-dose GLE group and cisplatin group according to different treatments, with 10 rats in each group. After the Walker-256 ascites rat model was successfully established, the rats in the low-dose, medium-dose and high-dose GLE groups were intragastrically administered with 0.84, 1.68 and 3.36 g/(kg·d) GLE, respectively, twice a day, 2 mL each time, for 14 days; the rats in the cisplatin group were intraperitoneally injected with 0.004 g/(kg·d) cisplatin in normal saline once a week; the rats in control group and model group were intragastrically administered with normal saline, 2 mL each time, twice a day for 14 days. The general health status of the rats of each group were observed, the mass of ascites was tested, the spleen, thymus and kidney indexes were calculated, and the renal function was measured. Flow cytometry was used to measure the cell cycles of thymus and bone marrow cells, the distributions of T lymphocyte subsets and the number of NK cells in peripheral blood. Serum immune cytokine protein chip was used to measure the serum levels of immune cytokines of rats in the control group, model group and medium-dose GLE group. Results GLE significantly reduced the mass of ascites in rats (P<0.05). Compared with the control group, thymus, spleen and kidney were damaged in the model group and cisplatin group, and treating rats with GLE significantly increased the thymus index and renal indexes, and significantly decreased serum creatinine and urea nitrogen (P<0.05). G0/G1 phase arrest was significantly induced in the thymus and bone marrow cells in the model group and the cisplatin group (P<0.05); and GLE significantly reduced the arrest of G0/G1 phase and significantly induced the transformation of thymus and bone marrow cells to G2/M and S phases (P<0.05). GLE significantly increased the number of CD4+ cells, the ratio of CD4+ to CD8+ cells and the number of NK cells, and significantly decreased the number of CD8+ cells in the peripheral blood of rats as compared with the model group (P<0.05). Compared with the model groups, the expressions of interferon-γ (IFN-γ), interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-10 and tumor necrosis factor-α (TNF-α) in serum of the rats in medium-dose GLE group were significantly increased (P<0.05). Conclusion GLE can effectively reduce the generation of ascites and improve immune function in Walker-256 ascites rats, and has no renal damage effect compared with cisplatin.
Key words:  Ganoderma lucidum  ascites  immunity  cytokines  T-lymphocytes  natural killer cells