化瘀破癥胶囊干预荷瘤小鼠血清肿瘤坏死因子活性的变化

Effect of huayu pozheng capsule on activities of serum tumor necrosis factor in tumor-bearing mice

目的:观察由有化瘀解毒、破癥散结功效药物组成的化瘀破癥胶囊对荷瘤小鼠肿瘤坏死因子活性的作用。方法:实验于1998-10/1999-10在成都中医药大学附属医院药理实验室完成,C57BL/6J小鼠50只,腋部皮下注射Lewis肺癌24h后,均分为5组,即厌氧棒菌苗组、荷瘤模型对照组、化瘀破癥胶囊4,2,1g/kg剂量组,每组10只。厌氧棒菌苗组按0.5g/mL腹腔注射,1次;荷瘤模型对照组灌胃等容积的蒸馏水,化瘀破癥胶囊(由四川省人民医院药剂科提供,由大黄、蜈蚣、人参等药组成,每粒装0.4g,相当于生药2g。)4,2,1g/kg剂量组分别按4,2,1g/kg给药,1次/d,连续7d,第8天停药,停药24h后每只鼠尾静脉注射脂多糖50g/L,2h后采血,分离血清,检测肿瘤坏死因子活性,并解剖小鼠取脾计算脾指数。脾指数=脾脏质量(mg)/体质量(g)。肿瘤坏死因子活性检测采用细胞毒试验法,用血球计数板计数200个肿瘤细胞中死细胞数,按给药组死细胞数/对照组死细胞数计算直接细胞毒活性指数表示肿瘤坏死因子活性,试验重复两批。结果:纳入实验小鼠50只,在实验过程中无小鼠死亡,进入结果分析50只。①化瘀破癥胶囊对荷瘤小鼠脾指数的影响:化瘀破癥胶囊4,2,1g/kg3个剂量组鼠的脾重均明显高于对照组(P<0.01),其脾指数两次试验,化瘀破癥胶囊4g/kg剂量组分别为1.69和1.47,2g/kg剂量组分别为1.57和1.66,1g/kg剂量组分别为1.65和1.46。两批试验与厌氧棒菌苗组比较,均未见明显差异(P>0.05)。②化瘀破癥胶囊对诱导产生肿瘤坏死因子活性的影响:两次试验结果,化瘀破癥胶囊4,2,1g/kg3个剂量组肿瘤细胞的死细胞数均显著高于对照组(P<0.01),其直接细胞毒指数,化瘀破癥胶囊4g/kg剂量组分别为2.29和2.12,2g/kg剂量组分别为2.36和2.05,1g/kg剂量组分别为2.18和2.09。并且两批试验与厌氧棒菌苗比较,均未见明显差异(P>0.05)。结论:化瘀破癥胶囊能显著增加荷瘤鼠脾脏质量和脾指数,以及增强血清中肿瘤坏死因子对肿瘤细胞的抑制杀伤活性,表明化瘀破癥胶囊对动物荷瘤机体的免疫功能有增强作用。

AIM： To investigate the effect of huayu pozheng capsule composed of stasis-dissipating and lump-removing herbs on activities of serum tumor necrosis factor in tumor bearing mice. METHODS： The experiment was conducted in the Laboratory for Pharmacology, the Affiliated Hospital of Cbengdu University of Traditional Chinese Medicine from October 1998 to October 1999. Fifty C57BL/6J mice were randomly divided into 5 groups with 10 mice in each group after 24 hours of armpit subcutaneous injection of Lewis lung cancer cells： anaerobic corynehacteria vaccine group received intraperitoneal injection of 0.5 g/mL eorynehacteria vaccine for once; tumor-bearing model group given gastric perfusion of distilled water with the same volume as above; 4, 2 and 1 g/kg dosage of huayu pozheng capsule groups received the huayu pozheng capsule （provided by the Department of Pharmacy, Sichuan Provincial People＇s Hospital, composed of dahuang, wugong, rencen and so on, 0.4 g per pill,the equal to 2 g crude drug） at dosage of 4, 2 and 1 g/kg, respectively,once a day for 7 days. On the 8^th day, the mice were quit of drug. Twentyfour hours later, the mice were given tail intravenous injection of 50 g/L lipopolysoccharide and taken blood after 2 hours to isolate the serum and detect TNF activities, then the spleen was taken to calculate the spleen index. Spleen index=spleen weight （mg）/body mass （g）. Cytotoxicity assay was used to detect TNF activity; blood cell counting chamber to count dead cells in 200 tumor cells; the directness cell toxin activity index denoted the TNF activities was calculated according to dead cell of the drug group/dead cell of the control group. This procedure was repeated twice. RESULTS： All the 50 mice were involved in the result analysis.①Effect of huayu pozheng capsule on spleen index in tumor-beating mice： The spleen weight in the 4, 2 and 1 g/kg dosage huayu pozheng capsule groups were all remarkable higher than the control group （P 〈 0.01）, and the spleen indexes in the two tests of the 4, 2 and 1 g/kg dosage huayu pozheng capsule groups were 1.69 and 1.47, 1.57 and 1.66, 1.65 and 1.46,respectively; and there was no significant difference between the two tests and the anaerobic corynebaeteria vaccine group （P 〉 0.05）. ②Effect of huayu pozheng capsule on activities of necrosis factor：. The dead cells of tumor cell in the 4, 2 and 1 g/kg dosage capsule groups were all remarkable higher than the control group （P 〈 0.01）, the directness eeU toxin activity indexes in the 4, 2 and 1 g/kg dosage capsule groups were 2.29 and 2.12, 2.36 and 2.05, 2.18 and 2.09, respectively; and there was no significant difference between the two tests and the anaerobic corynebacteria vaccine group （P 〉 0.05）. CONCLUSION： The huayu pozheng capsule can remarkably increase the spleen weight and spleen index, and enhance activities of serum TNF on inhibition and kill to tumor cells, indicating that the huayu pozheng capsule can enhance immune function of tumor-bearing organs.