莪术醇通过调控磷酯酰肌醇3激酶/蛋白激酶B信号通路对胶质瘤增殖与凋亡的影响  

Effect of curcomol on proliferation and apoptosis of glioma by regulating the phosphatidylinositol 3 kinase/protein kinase B signaling pathway

作　　者：张慧[1] 郑慧军[2] 杜康 韩庆亮 吴敬强 赵晓超 Zhang Hui;Zheng Huijun;Du Kang;Han Qingliang;Wu Jingqiang;Zhao Xiaochao(The Second Clinical Medical College of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China;Department of Neurosurgery,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南中医药大学第二临床医学院,郑州450000 [2]河南省中医院神经外科,郑州450002

出　　处：《中华实验外科杂志》2024年第4期732-737,共6页Chinese Journal of Experimental Surgery

基　　金：河南省中医药科学研究专项课题(2022ZY2014、2024ZY2090)。

摘　　要：目的探讨莪术醇通过磷酯酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号通路对神经胶质瘤增殖、凋亡的作用机制。方法不同浓度莪术醇(0、25、50、100、200μmol/L)干预人脑星形胶质母细胞瘤细胞(U87MG),观察细胞生长;细胞计数试剂盒(CCK-8)法检测细胞活力。将对数生长期U87MG细胞接种于裸鼠尾状核部位,建立胶质瘤原位模型。将模型裸鼠随机分组为模型组(等体积生理盐水)、莪术醇低剂量组(20 mg/kg)、中剂量组(40 mg/kg)和高剂量组(80 mg/kg),每组10只,连续灌胃治疗3周。观察各组小鼠行为变化;计算相对肿瘤体积、相对抑瘤率;苏木精-伊红(HE)染色观察瘤组织病理;免疫组织化学法(IHC)分析原位瘤增殖细胞核抗原(PCNA)表达及肿瘤细胞增殖指数;原位缺口末端标记法(TUNEL)染色分析肿瘤细胞凋亡水平;蛋白质印迹法(Western blot)分析PI3K/Akt信号通路相关蛋白及下游细胞周期蛋白D1(Cyclin D1)、B淋巴细胞瘤-2(bcl-2)、bcl-2相关X蛋白(bax)蛋白表达。两组间差异比较采用独立样本t检验,多组间差异比较采用One-way ANOVA分析。结果不同浓度莪术醇干预U87MG细胞后,随着给药浓度增加细胞出现漂浮碎片,凋亡细胞数增多;CCK-8结果显示,细胞存活率分别为对照组(1.00±0.07)%,25μmol/L组(1.10±0.04)%,50μmol/L组(0.67±0.04)%,100μmol/L组(0.58±0.06)%,200μmol/L组(0.24±0.01)%,各给药组的细胞存活率均低于对照组,差异有统计学意义(F=323.402,P<0.05)。计算瘤体体积及抑瘤率结果显示,模型组瘤体体积(42.50±27.78)mm^(3),各治疗组瘤体体积及抑瘤率分别为低剂量组[(31.85±20.89)mm^(3),25.20%],中剂量组[(21.49±13.42)mm^(3),49.54%],高剂量组[(13.73±12.0)mm^(3),67.75%],各治疗组瘤体体积均小于模型组,差异有统计学意义(F=2.147,P<0.05),高剂量组瘤体体积明显小于模型组[(13.73±12.05)mm^(3)比(42.50±27.78)mm^(3),t=2.130,P<0.05]。IHC结果显示,对照组、模型组、低剂量组、�Objective To explore the mechanism of curcumol on proliferation and apoptosis of glioma through phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt)signaling pathway.Methods Different concentrations of curcumol(0,25,50,100,200μmol/L)were used to intervene human astroblastoma cells(U87MG)and the cell growth was observed by researcher;In addition,the cell viability was tested by cell counting kit(CCK-8).Logarithmic U87MG cells were inoculated into caudate nucleus of nude mice which is contribute to establish glioma in situ model.In terms of the model nude mice,which were randomly divided into model group(equal volume of saline),low dose of curcumol group(20 mg/kg),middle dose of curcumol group(40 mg/kg)and high dose of curcumol group(80 mg/kg),10 mice in each group were treated by continuous gavage for 3 weeks.And then the researcher had an observation of the changes of the mice’s behavior.Calculate relative tumor volume and relative tumor inhibition rate;Histopathology of tumor was observed by hematoxylin-eosin(HE)staining;Immunohistochemical(IHC)analysis of proliferating cell nuclear antigen(PCNA)expression and tumor cell proliferation index;The apoptosis level of tumor cells was stained by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.Comparison of the expression of PI3K/Akt pathway related proteins and downstream cyclin D1,B cell lymphoma/lewkmia-2(bcl-2)and bcl-2 related X protein(bax)protein by Western blotting.The independent samples ttest was used for comparison of differences between the two groups,and one-way ANOVA analysis was used for comparison of differences between multiple groups.Results After applying the method of different concentrations of curcumol to intervene human astroblastoma cells(U87MG),and with the increasing of the the dosing concentration,floating fragments appeared in cells,besides,the number of apoptotic cells increasd at the same time.CCK-8 results show that,the cell survival rate were control group(1.00±0.07)%,25μmol/L group(1.10±0.04)%,5

关 键 词：胶质瘤 莪术醇 磷酯酰肌醇3激酶/Akt信号通路 增殖 凋亡 

分 类 号：R285[医药卫生—中药学]