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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
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王 芳 ,陆云阳 ,李韦韦 ,姚敏娜 (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
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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)
中图分类号 R965 文献标志码 A 文章编号 1001-0408(2023)14-1686-05
DOI 10.6039/j.issn.1001-0408.2023.14.05
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、64 μmol/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、8 μmol/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、8 μmol/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。
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
Effects of polyphyllin ⅥⅥ on the proliferation and apoptosis of glioma cells and potential mechanism
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WANG Fang ,LU Yunyang ,LI Weiwei ,YAO Minna (1. School of Medical Technology, Shaanxi Energy
Institute, Shaanxi Xianyang 712000, China;2. Dept. of Traditional Chinese Medicine and Natural
Pharmacology, Air Force Medical University, Xi’an 710032, China;3. Dept. of Pharmacy, Xijing Hospital,
Air Force Medical University, Xi’an 710032, China)
ABSTRACT OBJECTIVE To investigate the effects of polyphyllin Ⅵ(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, 64 μmol/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, 8 μmol/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, 8 μmol/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-3β protein 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 PPⅥ can 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-3β pathway.
KEYWORDS polyphyllin Ⅵ ; human glioma cell; proliferation; apoptosis; Fas/FasL death receptor pathway; Akt/GSK-3β
pathway
脑胶质瘤是一种源于神经胶质细胞的肿瘤,约占颅
Δ 基金项目 国家自然青年科学基金项目(No.81903862)
*第一作者 副教授,博士。研究方向:中药药理学。E-mail: 内肿瘤所有分型的 51% ,其恶性程度高、生长速度快、
[1]
wangfang3463704@163.com [2]
侵袭性和术后复发率高,患者的五年生存率低于10% ,
# 通信作者 副教授,硕士。研究方向:中药药理学。电话:029-
84775475。E-mail:yaona3698@163.com 是临床常见且较具有攻击性的原发性脑部恶性肿瘤之
· 1686 · China Pharmacy 2023 Vol. 34 No. 14 中国药房 2023年第34卷第14期