Objective To investigate the effects of ethanol extracts of Hedyotis Diffusae Herba (EEHDH) on in-vitro proliferation of human renal cancer 786-0 cells and growth of mouse renal cancer 786-0 cell tumor xenograft.Methods ①Renal cancer 786-0 cells were divided into the observation groups 1, 2, 3, 4, and the control group.Cells in the observation groups 1, 2, 3, and 4 were respectively added with the final concentrations of 10, 20, 40, and 80 mg/L EEHDH, while cells in the control group were added with the same volume of culture fluid.We measured the OD490 of 490 nm at 24, 48, and 72 h after culture by MTT.②Forty-two Kunming mice were randomly divided into 7 groups: the model group, the low-dose EEHDH group, medium-dose EEHDH group, high-dose EEHDH group, the cyclophosphamide group, medium-dose EEHDH combined with cyclophosphamide group, and control group with six in each group.Except that the mice in the control group were normally fed, the rest of 36 mice were injected with 0.2 mL tumor cell suspension subcutaneously at left hind leg axillae and weighed at 24 h.Mice in the model control group were given the same volume of drinking wate by gavage for 14 d.Mice in the low-, medium-, and high-dose EEHDH groups were given 5.0, 10.0, and 20.0 g/kg EEHDH, respectively, once a day, for 14 d.Mice in the cyclophosphamide group were given 20 mg/kg cyclophosphamide once every 2 days for 7 times.Mice in the medium-dose EEHDH combined with cyclophosphamide group were given 20 mg/kg CTX on day 1, 3, and 5, followed by 10.0 g/kg on day 6, once a day for 9 times.At 24 h after the last administration, we killed the mice to weigh and calculate the anti-tumor rate in each group.Then we separated the thymuses and spleens, weighed precisely, and calculated the thymus index and spleen index.Results ①Under the same conditions, the OD490 of renal cancer 786-0 cells in the observation groups 1, 2, 3, and 4 was lower than that in the control group (all P<0.05).The OD490 of renal cancer 786-0 cells was reduced with the increased duration of incubation and increased EEHDH dose (all P<0.05).②At the end of the experiment, the tumor weight in the model group, the low-dose EEHDH group, medium-dose EEHDH groups, high-dose EEHDH group, cyclophosphamide group, and medium-dose EEHDH combined with cyclophosphamide group was respectively (2.31±0.28), (1.94±0.22), (1.59±0.26), (1.52±0.23), (0.92±0.24), (0.86±0.23) g.The inhibitory rates of the low-dose EEHDH group, medium-dose EEHDH groups, high-dose EEHDH group, cyclophosphamide group, and medium-dose EEHDH combined with cyclophosphamide group were 16.02%, 31.22%, 34.26%, 60.20%, and 62.77%, respectively.The Q value of the median-dose EEHDH was 0.87.The mouse thymus index and spleen index of the model group were higher than those of the control group (both P<0.05).The mouse thymus index and spleen index of the thymophosphamide group were lower than those of the model group (both P<0.05).The spleen index of the low-, medium-, and high-doses EEHDH groups was higher than that of the model group (P<0.05), and no significant difference was found in the mouse thymus index as compared with that of the model group (P>0.05).The mouse thymus index and spleen index of the medium-dose EEHDH combined with cyclophosphamide group were higher than that of the cyclophosphamide group (both P<0.05).Conclusion EEHDH can significantly inhibit the proliferation of human renal carcinoma 786-0 cells in vitro and inhibit the growth of mouse renal cancer 786-0 cell transplantation tumor in vivo.through the enhancement of spleen function.%目的 探讨白花蛇舌草乙醇提取物(EEHDH)对人肾癌786-0细胞体外增殖及小鼠肾癌细胞786-0移植瘤生长的影响.方法 ①将人肾癌细胞786-0分为观察1、2、3、4组和对照组,观察1、2、3、4组分别加入终浓度为10、20、40、80 mg/L的EEHDH,对照组加入等体积培养液.培养24、48、72 h后采用MTT法于490 nm处测量吸光度值(OD490值).②取42只昆明小鼠,随机分为7组,分别为模型组、EEHDH低剂量组、EEHDH中剂量组、EEHDH高剂量组、环磷酰胺组、EEHDH中剂量联合环磷酰胺组、对照组,每组6只.除对照组小鼠正常饲养不处理外,余36只小鼠每只左后肢腋窝皮下接种0.2 mL肿瘤细胞悬液,24 h后称重.模型组灌胃等体积饮用水14 d;EEHDH低、中、高剂量组分别灌胃EEHDH 5.0、10.0、20.0 g/kg,1次/d,共14 d;环磷酰胺组每2 d灌胃1次20 mg/kg环磷酰胺, 共灌胃7次;EEHDH中剂量联合环磷酰胺组在1、3、5天灌胃环磷酰胺20 mg/kg, 第6天开始灌胃10.0 g/kg的EEHDH,1次/d,共9次.末次给药24 h后给各组小鼠称重,处死动物称瘤重,计算EEHDH低剂量组、EEHDH中剂量组、EEHDH高剂量组、环磷酰胺组、EEHDH中剂量联合环磷酰胺组抑瘤率.另分离各组小鼠胸腺、脾脏,精密称重,计算胸腺指数、脾脏指数.结果 ①相同培养时间条件下观察1、2、3、4组人肾癌细胞786-0的OD490值与对照组相比均较低(P均<0.05).人肾癌细胞786-0的OD490值随着培养时间的延长和EEHDH剂量的增加而降低(P均<0.05).②实验结束时,模型组、EEHDH低剂量组、EEHDH中剂量组、EEHDH高剂量组、环磷酰胺组、EEHDH中剂量联合环磷酰胺组瘤重分别为(2.31±0.28)、(1.94±0.22)、(1.59±0.26)、(1.52±0.23)、(0.92±0.24)、(0.86±0.23)g,EEHDH低剂量组、EEHDH中剂量组、EEHDH高剂量组、环磷酰胺组、EEHDH中剂量联合环磷酰胺组抑瘤率分别为16.02%、31.22%、34.26 %、60.20%、62.77%.EEHDH中剂量联合环磷酰胺Q 值为0.87.模型组小鼠胸腺指数、脾脏指数与对照组相比均较高(P均<0.05).环磷酰胺组小鼠胸腺指数、脾脏指数与模型组相比均较低(P均<0.05).EEHDH低、中、高剂量组小鼠脾脏指数与模型组相比均较高(P均<0.05),胸腺指数与模型组相比P均>0.05.EEHDH中剂量联合环磷酰胺组胸腺指数、脾脏指数与环磷酰胺组相比均较高(P均<0.05).结论 EEHDH在体外能明显抑制人肾癌细胞786-0的增殖,在体内能显著抑制小鼠肾癌细胞786-0移植瘤生长,其机制可能与EEHDH提高脾脏功能有关.
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