摘要
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。
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(Bcelllymphoma/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.
作者
施翠芬
谢斌
张汗顺
胡桥
刘成
余功
Shi Cuifen;Xie Bin;Zhang Hanshun;Hu Qiao;Liu Cheng;Yu Gong(The Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,China;College of Traditional Chinese Medicine,Jiangxi University of Chinese Medicine,Nanchang 330006,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2022年第7期2710-2716,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
江西省卫生和计划生育委员会中医药科研课题普通项目(2017A266):川楝子佐制配伍在一贯煎抗肝癌中的作用研究机制,负责人:余功
国家自然科学基金委员会地区科学基金项目(81660729):多靶点酶抑制剂?清燥救肺汤调控水形态影响肺癌细胞能量代谢的“清热润燥”机制研究,负责人:谢斌
江西省教育厅科学技术研究项目重点项目(GJJ201202):基于“能量剥夺诱导自噬”探索清燥救肺汤“清燥救肺”抗肺癌机制,负责人:谢斌
关键词
一贯煎
川楝子配伍
肝癌细胞凋亡
Yi Guan Decoction
Compatibility of Toosendan Fructus
Hepatoma cell apoptosis
