阿克替苷通过调控JNK信号通路促进肝癌细胞自噬与凋亡的实验研究  

Acteoside promotes autophagy and apoptosis of hepatoma cells by regulating JNK signaling pathway

作　　者：何昱静 郑英 李初谊 高浏璐 王俊科 李斌 卢利霞 王盼 于晓辉 张久聪 HE Yu-jing;ZHENG Ying;LI Chu-yi;GAO Liu-lu;WANG Jun-ke;LI Bin;LU Li-xia;WANG Pan;YU Xiao-hui;ZHANG Jiu-cong(Department of Gastroenterology,the 940th Hospital of Joint Logistics Support Force of Chinese People′s Liberation Army,Lanzhou 730050,China)

机构地区：[1]中国人民解放军联勤保障部队第九四〇医院消化内科,甘肃兰州730050

出　　处：《中国中药杂志》2023年第9期2343-2351,共9页China Journal of Chinese Materia Medica

基　　金：甘肃省非感染性肝病临床医学研究中心项目(21JR7RA017);临床医学研究中心培育项目(2021yxky079);甘肃省重点研发计划项目(20YF8FA099)。

摘　　要：探讨阿克替苷(acteoside)通过JNK信号通路抑制H22荷瘤小鼠瘤体的分子机制。50只BALB/c雄性小鼠经H22肝癌细胞皮下造模后,分为模型组、顺铂组以及阿克替苷低、中、高剂量组5组,每组均给药2周,每周连续给药5 d。观察各组小鼠精神、饮食、饮水、活动度、皮毛等一般状况。比较给药前后小鼠体质量变化及各组小鼠瘤体体积、瘤重和抑瘤率。HE染色观察肝癌组织形态学变化,免疫组化和Western blot检测各肿瘤组织中p-JNK、JNK、Bcl-2、Beclin-1和LC3的表达,qRT-PCR检测JNK、Bcl-2、Beclin-1和LC3 mRNA的表达。模型组和阿克替苷低剂量组小鼠一般状况较差,其余3组小鼠的一般状况均改善。阿克替苷中、高剂量组和顺铂组小鼠体质量小于模型组(P<0.01)。模型组瘤体体积与阿克替苷低剂量组比差异无统计学意义,顺铂组瘤体体积与阿克替苷高剂量组比差异无统计学意义。阿克替苷中、高剂量组和顺铂组瘤体体积和瘤重与模型组相比显著减小(P<0.001)。阿克替苷低、中、高剂量组和顺铂组的抑瘤率分别为10.72%、40.32%、53.79%、56.44%。HE染色显示,阿克替苷组和顺铂组肝癌细胞数逐渐减少,细胞坏死征象不断增多,阿克替苷高剂量组和顺铂组坏死区域较多。免疫组化结果显示,阿克替苷组和顺铂组可上调Beclin-1、LC3、p-JNK、JNK的表达(P<0.05)。免疫组化、Western blot和qRTPCR结果显示,阿克替苷中、高剂量组和顺铂组可下调Bcl-2的表达(P<0.01)。Western blot结果显示,阿克替苷组和顺铂组可上调Beclin-1、LC3和p-JNK的表达(P<0.01),各组JNK的表达水平差异无统计学意义。qRT-PCR结果显示,阿克替苷组和顺铂组可上调Beclin-1、LC3 mRNA的水平(P<0.05),阿克替苷中、高剂量组和顺铂组可上调JNK mRNA的水平(P<0.001)。综上,阿克替苷可上调JNK信号通路促进H22荷瘤小鼠肝癌细胞的凋亡和自噬进而发挥抑制瘤体生长的作用。This study explored the molecular mechanism of acteoside against hepatoma 22(H22)tumor in mice through c-Jun N-terminal kinase(JNK)signaling pathway.H22 cells were subcutaneously inoculated in 50 male BALB/c mice,and then the model mice were classified into model group,low-dose,medium-dose,and high-dose acteoside groups,and cisplatin group.The administration lasted 2 weeks for each group(5 consecutive days/week).The general conditions of mice in each group,such as mental status,diet intake,water intake,activity,and fur were observed.The body weight,tumor volume,tumor weight,and tumor-inhibiting rate were compared before and after administration.Morphological changes of liver cancer tissues were observed based on hematoxylin and eosin(HE)staining,and the expression of phosphorylated(p)-JNK,JNK,B-cell lymphoma-2(Bcl-2),Beclin-1,and light chain 3(LC3)in each tissue was detected by immunohistochemistry and Western blot.qRT-PCR was performed to detect the mRNA expression of JNK,Bcl-2,Beclin-1,and LC3.The general conditions of mice in model and low-dose acteoside groups were poor,while the general conditions of mice in the remaining three groups were improved.The body weight of mice in medium-dose acteoside group,high-dose acteoside group,and cisplatin group was smaller than that in model group(P<0.01).The tumor volume in model group was insignificantly different from that in low-dose acteoside group,and the volume in cisplatin group showed no significant difference from that in high-dose acteoside group.Tumor volume and weight in medium-dose and high-dose acteoside groups and cisplatin group were lower than those in the model group(P<0.001).The tumor-inhibiting rates were 10.72%,40.32%,53.79%,and 56.44%in the low-dose,medium-dose,and high-dose acteoside groups and cisplatin group,respectively.HE staining showed gradual decrease in the count of hepatoma cells and increasing sign of cell necrosis in the acteoside and cisplatin groups,and the necrosis was particularly obvious in the high-dose acteoside group and cisplati

关 键 词：阿克替苷 H22荷瘤小鼠 JNK信号通路 凋亡 自噬 

分 类 号：R285.5[医药卫生—中药学]