泰山山慈菇对小鼠肝癌的治疗作用及相关免疫机制研究(1)
摘 要：目的 探讨泰山山慈菇对小鼠肝癌的免疫治疗作用。方法 本课题首先制备泰山山慈菇提取液，体外用山慈菇提取液对H22肝癌细胞进行干预，分为空白组、山慈菇提取液低、中、高剂量组和5-氟尿嘧啶阳性对照组，干预12 h后，Annexin V和7-AAD染色，流式细胞术检测细胞凋亡。体内制备小鼠H22实体瘤肝癌模型，将50只小鼠分为山慈菇低、中、高剂量组、5-Fu阳性对照组、模型对照组，每组10只。山慈菇各剂量组均进行灌胃治疗，模型组灌胃等体积生理盐水，5-Fu阳性对照组进行等体积腹腔注射。连续 10 d，停药后次日摘取小鼠眼球取血后颈椎脱臼处死小鼠，检测小鼠肿瘤大小分析对肿瘤的抑制作用。酶联免疫吸附试验(ELISA)检测血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)和γ-干扰素(IFN-γ)的含量。结果 泰山山慈菇提取液体外促进H22肿瘤细胞凋亡且具有剂量依赖性。体内抑制H22肿瘤的生长，增加血清中TNF-α、IL-2和IFN-γ的含量且呈现剂量依赖性。结论 泰山山慈菇通过增加抗肿瘤细胞因子产生发挥抗肿瘤作用。
, http://www.100md.com 关键词：泰山山慈菇；肝癌；肿瘤坏死因子-α；白细胞介素-2；γ-干扰素
中图分类号：R392.3 文献标识码：A DOI：10.3969/j.issn.1006-1959.2018.09.003
文章编号：1006-1959(2018)09-0008-04
Abstract：Objective To investigate the immunotherapy effect of Taishan mountain mushroom on liver cancer in mice.Methods We first prepared the Taishan mountain mushroom extract.In vitro，the H22 hepatoma cells were treated with the extract of mushroom extract in vitro.It was divided into blank group，low，middle and high dose group and 5- fluorouracil positive control group.After 12 h intervention，Annexin V and 7-AAD were stained，and cell apoptosis was detected by flow cytometry.The mice model of H22 solid tumor was prepared in vivo，and 50 mice were divided into low，medium and high dose group，5-Fu positive control group and model control group，with 10 rats in each group.The rats in each dose group were treated by intragastric perfusion，and the model group was given iso-volume intraperitoneal injection of normal saline and 5-Fu positive control group.For 10 d，the mice were killed the next day after extirpation of eyeball and blood，and the inhibitory effect of tumor size analysis on the tumor was detected.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of TNF-α，IL-2 and IFN-γ in serum.Results The extract of Taishan mountain mushroom promoted the apoptosis of H22 tumor cells in a dose-dependent manner.In vivo，the growth of H22 tumor was inhibited and the levels of TNF-α，IL-2 and IFN-γ in serum increased.Conclusion Taishan mountain mushroom plays an anti-tumor role by increasing the production of anti-tumor cytokines.
, 百拇医药
Key words：Taishan mountain mushroom；Liver cancer；Tumor necrosis factor -α；Interleukin-2；γ-interferon
肝癌是死亡率較高、五年生存率较低、恶性程度极高的肿瘤疾病之一。肝癌的治疗主要有手术、放疗、化疗等治疗方法[1]。放疗、化疗对机体毒副作用较大，探讨中药对肝癌的治疗作用是目前研究的热点。山慈菇是常用的抗肿瘤中药之一。泰山山慈菇资源丰富，而其药用价值远远没有得到开发。据记载，山慈菇为中国药典收载的常用中药，中医临床上常用于治疗乳腺癌、食管癌、胃癌等恶性肿瘤等[2，3]，但其对于肝癌的作用效果鲜有研究，限制其在临床上的应用。其是否通过增强免疫作用抗肿瘤也不清楚。本研究首先采用醇提法制备泰山山慈菇提取液。体外观察泰山山慈菇提取液是否对H22细胞具有促凋亡作用，在此基础上制备小鼠H22实体瘤肝癌模型，体内检测其对H22荷瘤小鼠肿瘤生长的影响及对血清TNF-α、IL-2和IFN-γ含量的影响，探讨其可能的抗肿瘤免疫作用机制。, http://www.100md.com(修朋程 李腾 李雅林 刘纪豪 李栋 刘有旺)
, http://www.100md.com 关键词：泰山山慈菇；肝癌；肿瘤坏死因子-α；白细胞介素-2；γ-干扰素
中图分类号：R392.3 文献标识码：A DOI：10.3969/j.issn.1006-1959.2018.09.003
文章编号：1006-1959(2018)09-0008-04
Abstract：Objective To investigate the immunotherapy effect of Taishan mountain mushroom on liver cancer in mice.Methods We first prepared the Taishan mountain mushroom extract.In vitro，the H22 hepatoma cells were treated with the extract of mushroom extract in vitro.It was divided into blank group，low，middle and high dose group and 5- fluorouracil positive control group.After 12 h intervention，Annexin V and 7-AAD were stained，and cell apoptosis was detected by flow cytometry.The mice model of H22 solid tumor was prepared in vivo，and 50 mice were divided into low，medium and high dose group，5-Fu positive control group and model control group，with 10 rats in each group.The rats in each dose group were treated by intragastric perfusion，and the model group was given iso-volume intraperitoneal injection of normal saline and 5-Fu positive control group.For 10 d，the mice were killed the next day after extirpation of eyeball and blood，and the inhibitory effect of tumor size analysis on the tumor was detected.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of TNF-α，IL-2 and IFN-γ in serum.Results The extract of Taishan mountain mushroom promoted the apoptosis of H22 tumor cells in a dose-dependent manner.In vivo，the growth of H22 tumor was inhibited and the levels of TNF-α，IL-2 and IFN-γ in serum increased.Conclusion Taishan mountain mushroom plays an anti-tumor role by increasing the production of anti-tumor cytokines.
, 百拇医药
Key words：Taishan mountain mushroom；Liver cancer；Tumor necrosis factor -α；Interleukin-2；γ-interferon
肝癌是死亡率較高、五年生存率较低、恶性程度极高的肿瘤疾病之一。肝癌的治疗主要有手术、放疗、化疗等治疗方法[1]。放疗、化疗对机体毒副作用较大，探讨中药对肝癌的治疗作用是目前研究的热点。山慈菇是常用的抗肿瘤中药之一。泰山山慈菇资源丰富，而其药用价值远远没有得到开发。据记载，山慈菇为中国药典收载的常用中药，中医临床上常用于治疗乳腺癌、食管癌、胃癌等恶性肿瘤等[2，3]，但其对于肝癌的作用效果鲜有研究，限制其在临床上的应用。其是否通过增强免疫作用抗肿瘤也不清楚。本研究首先采用醇提法制备泰山山慈菇提取液。体外观察泰山山慈菇提取液是否对H22细胞具有促凋亡作用，在此基础上制备小鼠H22实体瘤肝癌模型，体内检测其对H22荷瘤小鼠肿瘤生长的影响及对血清TNF-α、IL-2和IFN-γ含量的影响，探讨其可能的抗肿瘤免疫作用机制。, http://www.100md.com(修朋程 李腾 李雅林 刘纪豪 李栋 刘有旺)