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目的探讨吉林双阳梅花鹿鹿茸蛋白组分对肿瘤细胞的细胞毒性作用。方法将新鲜鹿茸高速匀浆上清透析后,用超滤离心法获取分子量小于10kD,大于10kD小于30kD,大于30kD的3种蛋白组分;采用MTT法,检测3种蛋白组分对人肝癌SMMC-7721细胞系及人肺腺癌SPC-A-1细胞系的细胞毒性作用。另外,设鹿茸提取物原浓度的1/2,1/4,1/8为高,中,低3个浓度组,以0.01ml/g浓度皮下注射昆明小鼠,连续注射14天后,取小鼠脾细胞培养(培养基含ConA0.005mg/ml)72h后,加入MTT(5mg/ml)0.01ml,继续培养4h,测定OD值,计算淋巴细胞转化值。结果针对SMMC-7221细胞,分子量<10kD,>30kD,10kD<<30kD的鹿茸蛋白组分中,高剂量组的抑制率均大于30%,分子量<10kD,10kD<<30kD的鹿茸蛋白组分中,中高剂量组的抑制率大于30%,而且呈现剂量依赖性关系。针对SPC-A-1细胞,从所选取的高剂量至低剂量的抑制率均小于30%。鹿茸蛋白组分高、中、低剂量组的平均OD值分别为0.46±0.05、0.29±0.03和0.17±0.03;胸腺肽阳性对照组的平均OD值为0.47±0.07;生理盐水阴性对照组的平均OD值为0.12±0.04。鹿茸蛋白组分高、中、低剂量组的转化值分别为0.34、0.17和0.05;胸腺肽阳性对照组的转化值为0.35。鹿茸蛋白组分的高剂量组和胸腺肽对照组OD值比较,差异不具有显著性(P>0.05)。结论对于SPC-A-1细胞,鹿茸各蛋白组分无明显细胞毒性作用,而对于SMMC-7221细胞,各蛋白组分的高剂量、中高剂量组则呈现一定的细胞毒性作用。鹿茸蛋白组分高剂量组具有与胸腺肽注射液相近的促进淋巴细胞转化活性。
Objective To investigate the cytotoxic effect of pilose antler protein components from Jilin Shuangyang Sika deer on tumor cells. Methods The supernatant of fresh antler was dialyzed and the three protein fractions with molecular weight less than 10kD, greater than 10kD less than 30kD and greater than 30kD were obtained by ultrafiltration centrifugation. MTT assay was used to detect the expression of three protein components on human hepatoma SMMC -7721 cell line and human lung adenocarcinoma cell line SPC-A-1. In addition, the original concentration of antler velvet 1/2, 1/4, 1/8 for high, medium and low concentrations of 3 groups, 0.01 ml / g Kunming mice subcutaneously injected, continuous injection of 14 days, After cultured splenocytes (ConA0.005mg / ml) for 72h, 0.01ml of MTT (5mg / ml) was added and cultured for 4h. OD value was calculated and lymphocyte transformation was calculated. Results For SMMC-7221 cells, the inhibitory rates of high dose group were higher than 30%, molecular weight <10kD, 10kD << 30kD in antler protein components of molecular weight <10kD,> 30kD, 10kD << 30kD , The inhibition rate of high-dose group was more than 30%, and showed a dose-dependent relationship. For SPC-A-1 cells, the inhibition rates from high to low doses were less than 30%. The average OD value of high, medium and low dose group of antler protein components were 0.46 ± 0.05, 0.29 ± 0.03 and 0.17 ± 0.03 respectively; the average OD value of thymosin positive control group was 0.47 ± 0.07; the average OD of negative control group The value is 0.12 ± 0.04. The conversion values of antler protein components in high, medium and low dose groups were 0.34, 0.17 and 0.05, respectively; the conversion value of thymosin positive control group was 0.35. There was no significant difference in the OD value between the high-dose group and the thymosin control group (P> 0.05). Conclusion There is no cytotoxic effect on the protein components of antler in SPC-A-1 cells. However, SMMC-7221 cells show some cytotoxic effects in high and medium dose groups. The antler protein component high-dose group has the similar lymphocyte transformation activity as thymosin injection.