重楼皂苷Ⅵ对脑胶质瘤细胞增殖和凋亡的影响及潜在机制  

Effects of polyphyllinⅥon the proliferation and apoptosis of glioma cells and potential mechanism

作　　者：王芳 陆云阳 李韦韦 姚敏娜 WANG Fang;LU Yunyang;LI Weiwei;YAO Minna(School of Medical Technology,Shaanxi EnergyInstitute,Shaanxi Xianyang 712000,China;Dept.of Traditional Chinese Medicine and NaturalPharmacology,Air Force Medical University,Xi’an 710032,China;Dept.of Pharmacy,Xijing Hospital,Air Force Medical University,Xi’an 710032,China)

机构地区：[1]陕西能源职业技术学院医学技术学院,陕西咸阳712000 [2]空军军医大学中药与天然药物学教研室,西安710032 [3]空军军医大学西京医院药剂科,西安710032

出　　处：《中国药房》2023年第14期1686-1690,共5页China Pharmacy

基　　金：国家自然青年科学基金项目(No.81903862)。

摘　　要：目的探讨重楼皂苷Ⅵ(PPⅥ)对脑胶质瘤细胞增殖和凋亡的影响及潜在机制。方法以人脑胶质瘤LN229细胞为对象,采用MTT法检测不同浓度PPⅥ[0(对照组)、1、2、4、8、16、32、64μmol/L]作用不同时间(24、48、72 h)后的细胞存活率,采用克隆形成实验检测不同浓度PPⅥ[0(对照组)、2、4、8μmol/L]作用14 d后的细胞克隆数和克隆形成率,采用流式细胞术和Western blot法分别检测不同浓度PPⅥ[0(对照组)、4、8μmol/L]作用24 h后的细胞凋亡率和凋亡相关蛋白[B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、切割的胱天蛋白酶3(cleaved caspase-3)]、Fas/Fas配体(FasL)死亡受体通路相关蛋白、蛋白激酶B(又称Akt)/糖原合成激酶3β(GSK-3β)通路相关蛋白的表达情况。结果与对照组比较,PPⅥ各浓度组细胞的存活率、克隆数和克隆形成率、Bcl-2蛋白的表达水平和Akt、GSK-3β蛋白的磷酸化水平均显著降低(P<0.05或P<0.01);细胞凋亡率,Bax、cleaved caspase-3蛋白和Fas、FasL、cleaved caspase-8蛋白的表达水平均显著升高(P<0.05或P<0.01)。结论PPⅥ可抑制人胶质瘤LN229细胞的增殖并诱导其凋亡,这种作用可能与激活Fas/FasL死亡受体通路和抑制Akt/GSK-3β通路有关。OBJECTIVE To investigate the effects of polyphyllinⅥ(PPⅥ)on the proliferation and apoptosis of glioma cells and potential mechanism.METHODS Using human glioma LN229 cells as objects,MTT assay was used to detect the survival rate after treated with different concentrations of PPⅥ[0(control group),1,2,4,8,16,32,64μmol/L]for different time(24,48,72 h).The clone formation experiments were adopted to detect the number of cell clones and clone formation rate after being treated with different concentrations of PPⅥ[0(control group),2,4,8μmol/L]for 14 days.The flow cytometry and Western blot assay were used to detect the apoptotic rate of cells,the expressions of apoptosis-related protein[B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),cleaved caspase-3],and the expressions of related proteins of Fas/Fas ligand(FasL)death receptor pathway and protein kinase B(Akt)/glycogen synthesis kinase-3β(GSK-3β)pathway after being treated with different concentrations of PPⅥ[0(control group),4,8μmol/L]for 24 h.RESULTS Compared with the control group,the survival rate of cells,the number of clones and clone formation rate,the protein expression of Bcl-2,and the phosphorylation levels of Akt and GSK-3βprotein were decreased significantly in different concentration groups of PPⅥ(P<0.05 or P<0.01).The apoptotic rate,the protein expressions of Bax,cleaved caspase-3,Fas,FasL and cleaved caspase-8 were increased significantly(P<0.05 or P<0.01).CONCLUSIONS PPⅥcan inhibit the proliferation and induce the apoptosis of human glioma LN229 cells,which may be related to the activation of the Fas/FasL death receptor pathway and the inhibition of the Akt/GSK-3βpathway.

关 键 词：重楼皂苷Ⅵ 人脑胶质瘤细胞 增殖 凋亡 Fas/Fas配体死亡受体通路 蛋白激酶B/糖原合成激酶3β通路 

分 类 号：R965[医药卫生—药理学]