白附子提取物对胶质瘤细胞的增殖抑制和诱导凋亡作用及其机制

目的：研究白附子提取物对人SHG-44脑胶质瘤细胞生长的影响，初步探讨白附子对胶质瘤细胞增殖和凋亡的作用机制。方法：培养脑胶质瘤SHG-44细胞，将其分为空白对照组和8、40、200及1 000 μg•L^-1白附子提取物组，MTT法检测白附子提取物对SHG-44细胞生长的影响，倒置显微镜观察细胞凋亡形态，流式细胞术分析细胞周期和细胞凋亡率，细胞免疫组织化学法检测Bcl-2与Bax蛋白表达。结果：MTT结果，与空白对照组比较，8和200 μg•L^-1白附子提取物组细胞在30 min和3 h时，细胞增殖均明显受抑制，细胞增殖活性降低（P＜0.05)；1 000 μg•L^-1白附子提取物组细胞在30 min、1 h和3 h时，细胞增殖活性均明显降低（P＜0.05）；不同浓度白附子提取物对胶质瘤细胞增殖抑制作用呈剂量-时间依赖性。镜下观察，与空白对照组比较，各药物组细胞均出现数量不等的凋亡小体。流式细胞术分析，部分细胞阻滞于G2/M期，细胞凋亡率随着药物浓度的增加而升高。细胞免疫组织化学检测，与空白对照组比较，200 μg•L^-1白附子提取物组细胞Bcl-2蛋白表达降低（P<0.01），Bax蛋白表达增高（P<0.01）。结论：白附子提取物可抑制SHG-44细胞的增殖及诱导其发生凋亡，其作用机制与Bcl-2蛋白表达下降和Bax蛋白表达上升有关。
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Proliferation inhibition and inducing apoptosis of Rhizoma Typhonii extracts on glioma cells and their mechanisms

To study the influence of Rhizoma Typhonii extracts in glioma SHG-44 cells and to explore the effects of Rhizoma Typhonii extracts on proliferation and apoptosis of　glioma cells.Methods Glioma SHG-44 cells were cultured and divided into control and 8,40,200,and 1 000 μg•L^-1 Rhizoma Typhonii extracts groups,and the influence of Rhizoma Typhonii extracts in proliferation of SHG-44 cells was measured by MTT assay.The morphology of apoptosis was observed with inverted microscope.The cell cycle and apoptotic rate were analyzed by flow cytometry (FCM).The expressions of Bcl-2 and Bax protein were detected by immunohistochemistry.Results Compared with control group,the poliferation of SHG-44 cells was inhibited after treated with Rhizoma Typhonii extracts at concentrations of 8 and 200 μg•L^-1 for 30 min and 3 h and the proliferation activities were decreased(P＜0.05);in 1 000 μg•L^-1 group　after treated for 30 min,1 h and 3 h,the proliferation activities were significantly decreased (P＜0.05).The proliferation inhibition of differentconcentrations of Rhizoma Typhonii extracts was time- and dose-dependent.The apoptotic bodies were found under inverted microscope in different Rhizoma Typhonii extracts groups.The FCM results showed that part of cells were arrested at G2/M phase,and the apoptotic rates were increased with the increasing of drug concentrations.Cell immunohistochemisty showed that compared with control group the expression of Bcl-2 protein in 200 μg•L^-1 Rhizoma Typhonii extracts groups was decreased(P<0.01) and the expression of Bax protein was increased (P<0.01).Conclusion Rhizoma Typhonii extracts can inhibit the proliferation and induce apoptosis by down-regulating the Bcl-2 protein expression and up-regulating the Bax protein expression in SHG-44 cells.