川楝子配伍对一贯煎抑制荷H22小鼠肝癌细胞增殖的影响＊

目的 对比研究一贯煎,一贯煎去川楝子,川楝子对荷H22小鼠肝癌细胞增殖及凋亡相关蛋白B淋巴细胞瘤-2（B-celllymphoma/Leukemia-2,Bcl-2）、Bcl-2相关X蛋白（Bax）、细胞色素C（Cyt-c）、半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）蛋白表达的影响。方法 小鼠随机分为模型组,环磷酰胺（cyclophosphamide, CTX）组（50 mg·kg^-1）,一贯煎组（23 g·kg^-1）,一贯煎去川楝子组（21 g·kg^-1）,川楝子组（0.1 g·kg^-1）。雄性KM小鼠每组10只右侧腋下皮下注射H22细胞构建肝癌荷瘤模型,CTX组造模后腹腔注射给药,隔日一次,共7次,余组造模前后14天均灌胃给药,统计各组瘤重。原位末端标记法（TUNEL）检测肿瘤细胞凋亡;蛋白免疫印记法（Western blot）检测Bax、Bcl-2、Cyt-c、Caspase-3蛋白表达。结果 与模型组比较,CTX组,一贯煎组,一贯煎去川楝子组,川楝子组瘤重均降低（P<0.01）,肝癌细胞凋亡率均升高（P<0.01）,Bax、Caspase-3和Cyt-c蛋白表达均升高（P<0.05或P<0.01）,CTX组、一贯煎组Bcl-2蛋白表达降低（P<0.01）;与一贯煎组比较,一贯煎去川楝子组、川楝子组瘤重、Bcl-2蛋白表达升高（P<0.05）,肝癌细胞凋亡率和Bax、Caspase-3蛋白表达降低（P<0.05或P<0.01）;与一贯煎去川楝子组比较,川楝子组肝癌细胞凋亡率,Cyt-c、Caspase-3蛋白表达均升高（P<0.01）,Bcl-2蛋白表达降低（P<0.01）。结论 一贯煎中川楝子配伍能明显增强该方抑制肝癌细胞增殖的功效,其机理可能与升高Bax、Caspase-3和Cyt-c蛋白表达,降低Bcl-2蛋白表达,进而促进肝癌细胞凋亡有关。

Effects of Fructus Meliae Toosendan Compatibility in Yi Guan Decoction Apoptosis of H22 Tumor Bearing Mice

Objective To compare the effects of Yi Guan Decoction (YGD), YGD without Fructus Meliae Toosendan (FMT) group and FMT group on the expression of proliferation and apoptosis related proteins b-lymphocyte-2 (B-celllymphoma/Leukemia-2, Bcl-2), Bcl-2 related X protein (Bax), cytochrome c (Cyt-c) as well as Caspase-3 in H22 bearing mouse hepatoma cells.Methods Mice were randomly divided into model group (Normal saline, constant volume), cyclophosphamide (CTX) group (50 mg·kg^-1), YGD (23 g·kg^-1),YGD without FMT group (21 g·kg^-1), FMT group (0.1 g·kg^-1). 10 male KM mice in each groups were subcutaneously injected H22 cells into the right axilla to establish the tumor bearing model. Once every other day, CTX group was given intraperitoneal injection, for a total of 7 times. Before and after modeling, the other groups were given intragastric administration 14 days. Tumor weight of each group was counted; Tumor cell apoptosis was detected by in situ end labeling (TUNEL); The expressions of Bcl-2 associated X protein Bax, Cyt-c and Caspase-3 were detected by Western-blot.Results Compared with model group, the tumor weight of CTX group, YGD group, YGD without FMT group, FMT group was decreased (P<0.01). Apoptosis rate and protein expression of Bax, Caspase-3 and Cyt-c were significantly increased (P<0.05 or P<0.01). CTX group and YGD group of Bcl-2 protein expression was decreased (P<0.01). YGD without FMT group and FMT group tumor weight and Bcl-2 protein expression were higher than those YGD group (P<0.05). The apoptosis rate and the protein expression of Bax and Caspase-3 in hepatoma cells were lower than those YGD group (P<0.05 or P<0.01). YGD without FMT group of Cyt-c protein was lower than YGD group (P<0.01). FMT group of apoptosis rate and the expression of Cyt-c and Caspase-3 in hepatocellular carcinoma cells were higher than those in YGD without TF group (P<0.01). FMT group of expression of Bcl-2 protein was lower than that in YGD without TF group (P<0.01).Conclusions Toosendan Fructus compatibility in YGD could enhance the inhibitory effect on the proliferation of in H22 mice. The mechanism may be related to promoting Bax, Caspase-3, and Cyt-c protein expression and inhibiting Bcl-2 protein expression.