白术多糖抗神经细胞缺氧性凋亡的机制研究  

Mechanism of Atractylodes macrocephalaon polysaccharides against neuron apoptosis induced by hypoxia /reoxygenation

作　　者：胡微煦[1] 向勤[2] 文珠 何丹[4] 夏晓健[4] 胡国柱[4] 

机构地区：[1]复旦大学附属中山医院,上海200032 [2]南昌大学研究生院医学部,南昌330006 [3]江西省医学科学研究院,南昌330006 [4]江西省人民医院,南昌330006

出　　处：《中药药理与临床》2013年第4期84-88,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：江西省科技厅社会发展计划(2007BS22602)

摘　　要：目的:探讨白术多糖抗神经细胞缺氧性凋亡的机制。方法:研究分为正常对照组、凋亡阳性组、白术多糖0.025g/L组、白术多糖0.05g/L组、白术多糖0.1g/L组、白术多糖0.25g/L组。采用"Neurobasal+B27"体外神经细胞无血清培养,免疫细胞化学鉴定神经细胞,MTT测定药物毒性,Rh-123染色流式细胞仪检测线粒体损伤,RT-PCR及免疫细胞化学测定Caspase-3、Bax、Bcl-2 mRNA及蛋白表达。结果:与凋亡阳性组比较,白术多糖在0.025g/L^0.1g/L降低缺氧的神经细胞线粒体损伤,0.025g/L^0.05g/L下调缺氧的神经细胞Caspase-3 mRNA的表达,0.05g/L下调缺氧的神经细胞Bax mRNA的表达,0.025g/L^0.25g/L下调缺氧的神经细胞Caspase-3和Bax蛋白表达,0.025g/L^0.25g/L上调缺氧的神经细胞Bcl-2蛋白表达,0.025g/L^0.05g/L提高Bcl-2/Bax mRNA及0.025g/L^0.1g/L提高Bcl-2/Bax蛋白的比例(P<0.05)。结论:白术多糖在一定发范围内能有效地抑制神经细胞缺氧性凋亡,其机理是降低凋亡基因及蛋白产生,上调抗凋亡蛋白产生,提高Bcl-2/Bax比例。Objective： To explore the mechanism of Atractylodes macrocephalaon polysaccharides on preventing the neuron apoptosis indued by hypoxia.Methods： The study was divided into normal control group,apoptosis positive group induced by hypoxia,experiment group 1 treated by 0.025g / L Atractylodes macrocephalaon polysaccharides,experiment group 2 treated by 0.05g / L Atractylodes macrocephalaon polysaccharides,experiment group 3 treated by 0.1g / L Atractylodes macrocephalaon polysaccharides,experiment group 4 treated by 0.25 g / L Atractylodes macrocephalaon polysaccharides.Neurons were cultured by Neurobasal ＋ B27 in serum-free in vitro.Neurons were identified by immunocytochemistry.Drug toxicity was determined by MTT assay.Mitochondria injury was examined by Rhodamine 123 staining in flow cytometry.The mRNA and protein expressions of Caspase-3,Bax and Bcl-2 were identified by RT-PCR and immunocytochemistry.Results： The mitochondria injury was significantly inhibited by Atractylodes macrocephalaon polysaccharides from 0.025g / L to 0.1g / L in hypoxic neurons,the level of caspase-3 mRNA was significantly down-regulated by Atractylodes macrocephalaon polysaccharides from 0.025g / L to 0.05g / L in hypoxic neurons,the level of Bax mRNAs was significantly down-regulated by Atractylodes macrocephalaon polysaccharides at 0.05g / L in hypoxic neurons,the levels of caspase-3 and Bax proteins were significantly down-regulated by Atractylodes macrocephalaon polysaccharides from 0.025g / L to 0.25g / L in hypoxic neurons,the level of Bcl-2 protein was significantly up-regulated by Atractylodes macrocephalaon polysaccharides from 0.025g / L to 0.25g / L in hypoxic neurons and the ratioes of Bcl-2 / Bax from 0.025g / L to 0.05g / L in mRNA and from 0.025g / L to 0.1g / L in protein were significantly elevated by Atractylodes macrocephalaon polysaccharides in hypoxic neurons as compared to the apoptosis positive group（P ＆lt; 0.05）.Conclusion： The Atractylodes macrocephalaon polysaccharides has an effect on inhibitin

关 键 词：白术多糖 神经细胞 缺氧 凋亡 半胱氨酸天冬氨酸蛋白酶-3(Caspase-3) B淋巴细胞瘤-2基因相关蛋白(BCL-2-asso-ciated X protein Bax) B淋巴细胞瘤-2基因(Bcl-2)x 

分 类 号：R285[医药卫生—中药学]