玉屏风散对荷肝癌小鼠顺铂干预后免疫和脏器损伤的影响
摘要
目的：探讨扶正方药玉屏风散对肝癌小鼠顺铂干预后免疫和脏器的影响及其作用特点。
方法：脾细胞体外增殖实验,观察不同浓度玉屏风散对正常小鼠脾细胞体外正常培养及ConA刺激下增殖的影响。C57BL-6小鼠皮下接种2×106个Hepαl-6肝癌细胞建立肝癌小鼠模型,观察：1)各组肝癌模型小鼠生活状态,脾、胸腺指数及抑瘤率,分析不同剂量顺铂对肝癌小鼠肿瘤和免疫系统的影响；2)各组肝癌模型小鼠生活状态,脾、胸腺指数,体外脾淋巴细胞增殖,脾淋巴细胞及其细胞亚群CD4+、CD8+、CDllb+比例,和抑瘤率,分析不同剂量玉屏风散对肝癌小鼠肿瘤和免疫系统的影响；3)通过上述指标确定玉屏风散、顺铂的联用剂量,进一步观察二者联用的效应变化及作用特点,包括肾脏、肝脏、胃和肺重量,超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量、组织病理观察等指标。
结果：1)顺铂可抑制Hepal-6肝癌小鼠的体重、肿瘤生长,具有剂量依赖性,5mg/kg剂量基本达到抑瘤作用平台期,抑瘤率为74.45%。顺铂对免疫器官(脾脏和胸腺)具有损伤作用,2.5mg/kg剂量即可降低其重量和相应的脏器指数,具有剂量依赖性。2)玉屏风散对Hepal-6肝癌小鼠的肿瘤生长有一定的抑制效应,具有剂量依赖性,实验最大剂量(30g/kg)的抑制率为32.14%。对体外培养的脾细胞有促进增殖的作用；对肝癌小鼠的免疫器官(脾脏和胸腺)无影响,对脾CD4+和CD8+T细胞比例无影响,但能提高脾淋巴细胞和巨噬细胞比例。3)玉屏风散(25g/kg)与顺铂(2.5mg/kg)联用,其抑瘤作用为两者的相加效应；在肿瘤状态下,可减少顺铂对脾脏和胸腺的损伤；改善顺铂引起的脾淋巴细胞和巨噬细胞比例的减少；纠正顺铂引起的脾CD4+和CD8+T细胞比例非正常性的增高；并能减轻肝癌小鼠本身和顺铂对其肾脏、肝脏、胃、肺主要脏器的损伤。
结论：扶正方药玉屏风散对肝癌小鼠顺铂干预后有增效减毒作用,其特点是能保护肝癌本身致小鼠及顺铂致肝癌小鼠免疫系统和主要脏器的损伤,提高免疫调节功能和抗氧化是玉屏风散发挥作用的可能机制。
OBJECTIVE:To investigate the protection of Yupingfeng Decoction (YPF) on immune system and organ after the liver cancer-bearing mice treated with cispaltin (DDP)
METHODS:Separated primary splenic cells from healthy C57mice and cultured in the nutrient medium that contained the different concentrations of YPF or YPF conbined ConA. The cell proliferation was detected by MTT assay. Established the hepatoma model by injecting 2xlO6 Hepal-6 cells into C57 mice intradermally. Then:1) Evaluated the changes of tumor and immune system of these cancer-bearing mice which treated with different doses of DDP by observing the healthy condition, tumor weight, the spleen index and the thymus index.2) Evaluated the changes of tumor and immune system of these cancer-bearing mice which treated with different doses of YPF by observing the living condition, tumor weight, the spleen index and the thymus index, the activity of splenic cell proliferation in vitro, the percentage of splenic lymphocyte and CD4+n CD8+、CDllb+subgroups.3) The liver cancer-bearing mice were treated with YPF and DDP. And the combinational dose of DDP or YPF was definited by the parameters above-mentioned. Beside these parameters above-mentioned, also measured and analyzed the weight of kidney, liver, stomach and lung, detected the activity of Superoxide Dismutase (SOD), malondialdehyde (MDA), and observed pathologic changes.
RESULTS:1) DDP had a dose-dependent effect on inhibiting the weight and the growth of tumor.5mg/kg was the dose of the plateau phase of its inhibition effect, which tumor inhibition rate is 74.45%. DDP also can damage the immune organs (spleen and thymus) in dose-dependent manner, and 2.5mg/kg began shows the damage effect.2) YPF had a dose-dependent effect on inhibiting the growth of tumor, and the inhibition rate of its experiment maximal dose (30g/kg) was 32.14%. YPF could promote spleen cell proliferation in vitro, but had no significant effects on the immune organs (spleen and thymus) and the proportion of CD4+and CD8+T cell, nevertheless, YPF could elevate the proportion of splenic lymphocyte and macrophagocyte.3) The tumor inhibition showed an addition effect when combine DDP (2.5mg/kg) with YPF (25g/kg). This combinational therapy could relieve the damages on immune organs, improve the decrease of splenic lymphocyte and macrophagocyte, and correct the abnormal increase of CD4-and CD8+T cell in the liver-cancer-bearing mice. In addition, the combinational therapy also can protect the organ (kidney, liver, stomach, lung) from the damage that induced by tumor and the therapy of DDP.
CONCLUSIONS:YPF has a synergism and attenuation effect on the liver-cancer-bearing mice treated with DDP. YPF can protect the immune system and the organs from the damage that induced by tumor and DDP. The possible mechanism of these effects is that YPF can enhance immune function and has the activity of antioxdant.
方法：脾细胞体外增殖实验,观察不同浓度玉屏风散对正常小鼠脾细胞体外正常培养及ConA刺激下增殖的影响。C57BL-6小鼠皮下接种2×106个Hepαl-6肝癌细胞建立肝癌小鼠模型,观察：1)各组肝癌模型小鼠生活状态,脾、胸腺指数及抑瘤率,分析不同剂量顺铂对肝癌小鼠肿瘤和免疫系统的影响；2)各组肝癌模型小鼠生活状态,脾、胸腺指数,体外脾淋巴细胞增殖,脾淋巴细胞及其细胞亚群CD4+、CD8+、CDllb+比例,和抑瘤率,分析不同剂量玉屏风散对肝癌小鼠肿瘤和免疫系统的影响；3)通过上述指标确定玉屏风散、顺铂的联用剂量,进一步观察二者联用的效应变化及作用特点,包括肾脏、肝脏、胃和肺重量,超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量、组织病理观察等指标。
结果：1)顺铂可抑制Hepal-6肝癌小鼠的体重、肿瘤生长,具有剂量依赖性,5mg/kg剂量基本达到抑瘤作用平台期,抑瘤率为74.45%。顺铂对免疫器官(脾脏和胸腺)具有损伤作用,2.5mg/kg剂量即可降低其重量和相应的脏器指数,具有剂量依赖性。2)玉屏风散对Hepal-6肝癌小鼠的肿瘤生长有一定的抑制效应,具有剂量依赖性,实验最大剂量(30g/kg)的抑制率为32.14%。对体外培养的脾细胞有促进增殖的作用；对肝癌小鼠的免疫器官(脾脏和胸腺)无影响,对脾CD4+和CD8+T细胞比例无影响,但能提高脾淋巴细胞和巨噬细胞比例。3)玉屏风散(25g/kg)与顺铂(2.5mg/kg)联用,其抑瘤作用为两者的相加效应；在肿瘤状态下,可减少顺铂对脾脏和胸腺的损伤；改善顺铂引起的脾淋巴细胞和巨噬细胞比例的减少；纠正顺铂引起的脾CD4+和CD8+T细胞比例非正常性的增高；并能减轻肝癌小鼠本身和顺铂对其肾脏、肝脏、胃、肺主要脏器的损伤。
结论：扶正方药玉屏风散对肝癌小鼠顺铂干预后有增效减毒作用,其特点是能保护肝癌本身致小鼠及顺铂致肝癌小鼠免疫系统和主要脏器的损伤,提高免疫调节功能和抗氧化是玉屏风散发挥作用的可能机制。
OBJECTIVE:To investigate the protection of Yupingfeng Decoction (YPF) on immune system and organ after the liver cancer-bearing mice treated with cispaltin (DDP)
METHODS:Separated primary splenic cells from healthy C57mice and cultured in the nutrient medium that contained the different concentrations of YPF or YPF conbined ConA. The cell proliferation was detected by MTT assay. Established the hepatoma model by injecting 2xlO6 Hepal-6 cells into C57 mice intradermally. Then:1) Evaluated the changes of tumor and immune system of these cancer-bearing mice which treated with different doses of DDP by observing the healthy condition, tumor weight, the spleen index and the thymus index.2) Evaluated the changes of tumor and immune system of these cancer-bearing mice which treated with different doses of YPF by observing the living condition, tumor weight, the spleen index and the thymus index, the activity of splenic cell proliferation in vitro, the percentage of splenic lymphocyte and CD4+n CD8+、CDllb+subgroups.3) The liver cancer-bearing mice were treated with YPF and DDP. And the combinational dose of DDP or YPF was definited by the parameters above-mentioned. Beside these parameters above-mentioned, also measured and analyzed the weight of kidney, liver, stomach and lung, detected the activity of Superoxide Dismutase (SOD), malondialdehyde (MDA), and observed pathologic changes.
RESULTS:1) DDP had a dose-dependent effect on inhibiting the weight and the growth of tumor.5mg/kg was the dose of the plateau phase of its inhibition effect, which tumor inhibition rate is 74.45%. DDP also can damage the immune organs (spleen and thymus) in dose-dependent manner, and 2.5mg/kg began shows the damage effect.2) YPF had a dose-dependent effect on inhibiting the growth of tumor, and the inhibition rate of its experiment maximal dose (30g/kg) was 32.14%. YPF could promote spleen cell proliferation in vitro, but had no significant effects on the immune organs (spleen and thymus) and the proportion of CD4+and CD8+T cell, nevertheless, YPF could elevate the proportion of splenic lymphocyte and macrophagocyte.3) The tumor inhibition showed an addition effect when combine DDP (2.5mg/kg) with YPF (25g/kg). This combinational therapy could relieve the damages on immune organs, improve the decrease of splenic lymphocyte and macrophagocyte, and correct the abnormal increase of CD4-and CD8+T cell in the liver-cancer-bearing mice. In addition, the combinational therapy also can protect the organ (kidney, liver, stomach, lung) from the damage that induced by tumor and the therapy of DDP.
CONCLUSIONS:YPF has a synergism and attenuation effect on the liver-cancer-bearing mice treated with DDP. YPF can protect the immune system and the organs from the damage that induced by tumor and DDP. The possible mechanism of these effects is that YPF can enhance immune function and has the activity of antioxdant.
引文
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