白花蛇舌草提取物对肝癌细胞JNK/p38MARK通路及细胞学行为的影响  

Effects of hedyotis diffusa extract on JNK/p38MARK pathway and cytological behavior of hepatoma cells

作　　者：刘皎皎 李粉萍 杨跃青 何瑾瑜 曹雪艳 薛敬东 叶苗青 LIU Jiaojiao;LI Fenping;YANG Yueqing;HE Jinyu;CAO Xueyan;XUE Jingdong;YE Miaoqing(Shaanxi Hospital of Traditional Chinese Medicine,Xi'an,Shaanxi 710003,China)

机构地区：[1]陕西省中医医院肝病科,陕西西安710003

出　　处：《安徽医药》2022年第8期1496-1500,I0001,共6页Anhui Medical and Pharmaceutical Journal

基　　金：国家自然科学基金(81373513);2018年国家中医药管理局区域中医(肝病)诊疗中心培育单位建设项目[国中医药办医政函(2017)39号]。

摘　　要：目的研究白花蛇舌草提取物(HDE)对肝癌QGY细胞增殖、凋亡的影响,并探讨其抗肿瘤的可能作用机制。方法体外培养肝癌QGY细胞,分为对照组:RPMI-1640培养基培养;阳性对照组:全反式维甲酸(AT-RA)(5μmol/L);白花蛇舌草提取物(HDE)低、中、高剂量组:分别给予20、40、80 g/L HDE。四甲基偶氮唑盐比色法(MTT)检测细胞增殖能力,采用Hoechst33258荧光染色以及流式AnnexinV-FITC/PI双染法检测细胞凋亡率;通过蛋白质印迹法(Western blotting)检测增殖相关蛋白增殖细胞核抗原(PCNA)、凋亡相关蛋白B淋巴细胞瘤基因-2(Bcl-2)、Bcl-2相关X蛋白基因(Bax)、半胱氨酰天冬氨酸特异性蛋白酶3(caspase3)及JNK/p38MARK通路相关蛋白水平。结果24、48 h时QGY细胞增殖抑制率,与对照组[(16.56±3.15)%、(19.27±3.33)%]同时间点比较,20、40、80 g/L HDE组[24 h(33.37±11.36)%、(47.57±13.62)%、(59.37±16.95)%,48 h(36.56±10.03)%、(50.59±13.87)%、(63.61±16.99)%]、阳性对照组[(60.43±17.09)%、(64.63±17.15)%]均显著升高(P<0.05)。与对照组比较,20、40、80 g/L浓度HDE处理后QGY细胞凋亡率、Bax、caspase-3蛋白均显著升高(P<0.05),PCNA、Bcl-2蛋白、p-JNK/JNK、p-p38/p38表达降低(P<0.05),且均呈浓度依赖性,80 g/L HDE组细胞凋亡率、Bax、caspase-3、PCNA、Bcl-2蛋白、p-JNK/JNK、p-p38/p38表达与阳性对照组比较差异无统计学意义(P>0.05)。结论HDE可抑制肝癌QGY细胞增殖,促进QGY细胞凋亡,可能是通过抑制JNK/p38MARK通路活化进而发挥抗肿瘤作用。Objective To study the effects of hedyotis diffusa extract(HDE)on the proliferation and apoptosis of hepatoma QGY cells,and to explore the possible mechanism of its anti-tumor effect.Methods The QGY cells were cultured in vitro and divided into control group:cultured in RPMI-1640 culture medium;positive control group:all trans retinoic acid(AT-RA)(5μmol/L);low,medium and high dose groups of Hedyotis diffusa extract(HDE):20,40 and 80 g/L HDE respectively.MTT method was used to detect cell proliferation,Hoechst33258 fluorescence staining and flow Annexin V-FITC/PI double staining were used to detect the apoptosis rate;in addition,the levels of proliferating cell nuclear antigen(PCNA),B-lymphoma gene-2(Bcl-2),Bcl-2 related X protein gene(Bax),caspase-3 and JNK/p38MARK pathway related proteins were detected by Western blotting.Results Compared with those in the control group[(16.56±3.15)%,(19.27±3.33)%]at the same time point,the inhibition rate of QGY cell proliferation in 20,40,80 g/L HDE groups[24 h(33.37±11.36)%,(47.57±13.62)%,(59.37±16.95)%,48 h(36.56±10.03)%,(50.59±13.87)%,(63.61±16.99)%]and positive control group[(60.43±17.09)%,(64.63±17.15)%]at 24,48 h increased significantly(P<0.05).Compared with those in the control group,the apoptosis rate,Bax and caspase-3 protein of QGY cells after treatment with HDE with concentration of 20,40 and 80 g/L increased significantly(P<0.05),the expressions of PCNA,Bcl-2 protein,p-JNK/JNK,p-p38/p38 decreased(P<0.05),and all of them were concentration dependent,while there was no significant difference in apoptosis rate,Bax,Caspase-3,PCNA,Bcl-2 protein,p-JNK/JNK,p-p38/p38 expressions between the 80 g/L HDE group and the positive control group(P>0.05).Conclusion HDE can inhibit the proliferation of QGY cells of liver cancer and promote the apoptosis of QGY cells,which may play an anti-tumor role by inhibiting the activation of JNK/p38MARK pathway.

关 键 词：白花蛇舌草提取物 肝肿瘤 c-Jun氨基末端激酶/p38丝裂原活化蛋白激酶通路 QGY细胞株 增殖 凋亡 

分 类 号：R285[医药卫生—中药学]