楚丽丽,宛蕾
CHU Lili,WAN Lei

• 1:贵州医科大学基础医学院药理教研室

摘要(Abstract)：

目的:研究人参皂苷Rd对U251神经胶质瘤细胞凋亡的影响。方法:U251细胞分空白对照组和低、中、高剂量人参皂苷Rd组,分别与U251细胞作用48 h后,采用Hoechst33258荧光染色观察细胞凋亡形态、流式细胞仪Annexin-V FITC/PI双染检测细胞凋亡率、PI染色检测细胞周期、免疫组化法检测细胞周期蛋白CyclinD1的表达。结果:Hoechst33258荧光染色表明各给药组均可见细胞凋亡的形态特征,Annexin-V FITC/PI双染结果示人参皂苷Rd可诱导U251细胞凋亡,且有剂量依赖性;PI染色结果显示人参皂苷Rd阻滞细胞于G0/G1期,免疫组化结果显示人参皂苷Rd可下调CyclinD1蛋白表达。结论:Rd可体外诱导U251细胞凋亡,并抑制细胞周期进程,其机制可能与下调CyclinD1蛋白表达有关。
Objective: To study the effect of ginsenoside Rd on apoptosis of glioma U251 cells.Methods: U251 cells were treated with blank(control group) or ginsenoside Rd(low, medium and high-dose groups). At 48 hr after the treatment, the cells were stained with Hoechst 33258 fluorescence to detect the morphology of apoptotic cells, stained with Annexin-V FITC apoptotic rates, and stained with PI to detect the cell cycle. Immunohistochemistry was used to detect the expression of CyclinD1.Results: Hoechst 33258 fluorescence staining showed that the morphological characteristics of apoptotic cells were visible in each ginsenoside Rd group. Annexin-V FITC manner. PI staining revealed that ginsenoside Rd blocks cells at the G0 analysis showed that ginsenoside Rd downregulated CyclinD1 protein expression.Conclusion:Ginsenoside Rd induced apoptosis of U251 cells and inhibited cell cycle progression. The mechanism may be related to the downregulation of CyclinD1 protein expression.

关键词(KeyWords)： 中草药;人参属;细胞系,肿瘤;细胞周期;细胞凋亡;人参皂苷
drugs,Chinese herbal;panax;cell line,tumor;cell;apoptosis;ginsenoside

Abstract:

Keywords:

基金项目(Foundation): 贵州省科学技术基金项目[黔科合J字(2005)2070]

作者(Author): 楚丽丽,宛蕾
CHU Lili,WAN Lei

DOI: 10.19367/j.cnki.1000-2707.2020.02.010

参考文献(References)：

• [1]刘德育，曾飒．从三期药材中快速批量分离提取人参皂苷Rd的方法[J]．中药材，2006,29(3):247-249.
• [2] YE R D,LI N L,HAN J L,et al. Neuroprotective effects of ginsenoside Rd against oxygen-glucose deprivation in cultured hippocampal neurons[J]. Neurosience Research,2009,64(10):306-310.
• [3] YANG L,DENG Y H,XU S J,et al. In vivo pharmacokinetic and metabolism studies of ginsenoside Rd[J]. Journal of Chromatography B, 2007,854:77-84.
• [4] YE R D,HAN J L,KONG X W,et al. Protective effects of ginsenoside Rd on PC12 cells against HydrogenPeroxide[J]. Biol. Pharm. Bull, 2008,31(10):1923-1927.
• [5] LI X Y,LIANG J,TANG Y B,et al. Ginsenoside Rd prevents glutamate-induced apoptosis in rat cortical neurons[J]. Clinical and Experimental Pharmacology and Physiology, 2010,37:(2)199-204.
• [6] LIU X,WANG L,WEN A,et al. Ginsenoside-Rd improves outcome of acute ischaemic stroke-a randontized,doubleblind,placebo-controlled,multicenter trial[J]. European Journal of Neurology, 2012,19(6):855-863.
• [7] GUAN Y Y,ZHOU J G,ZHANG Z,et al. Ginsenoside-Rd from panax notoginseng blocks Ga2+influx through receptor-and store-operated Ga2+channels in vascular smooth muscle cells[J]. European Journalof Pharmacology,2006,548(3):129-136.
• [8] ZHANG X,SHI M,YE R,et al. Ginsenoside Rd altenuates tau protein phosphorylation via the PI3K/AKT/GSK-3beta pathway after transient forebrain ischemia[J]. Neurochemical Research, 2014,39(7):1363-1373.
• [9] LIU J,YAN X,LING L,et al. Ginsenoside Rd improves learning and memory ability in APP transgenic mice[J].Journal of Molecular Neuroscience, 2015,57(4):522-528.
• [10]YANG X L,GUO T K,WANG Y H,et al. Therapeutic effect of ginsenoside Rd in rats with TNBS. induced recurrent ulcerative colitis[J]. Archives ofPharmacai Research, 2012,35(7):1231-1239.
• [11]李小勇，刘雅．人参皂苷Rd抑制肝星状细胞活化作用与机制研究[J]．中国药理与临床，2017,33(2):34-37.
• [12]CHOI S S,LEE J K,SUH H W. Effect of ginsenosides administered intrathecally on the antinociception induced by cold water swimming stress in the mouse[J]. Biol Pharm Bull, 2003,26(6):858-861.
• [13]屠令锋，曹荣，段丽，等．人参皂甙Rd对SNI大鼠痛觉异常及脊髓背角内谷氨酸和NR2A/B表达的影响[J]．神经解剖学杂志，2011,27(2):129-134.
• [14]曹荣，屠令峰，段丽，等．人参皂甙Rd对SNI大鼠痛敏异常脊髓背角内P物质和NK-1受体表达的影响[J]．神经解剖学杂志，2011,27(1):8-13.
• [15]OSTROM Q T,GITTLEMAN H,FARAH P,et al. Cbtrus statistical report:pri-mary brain and central nervous system tumors diagnosed in the United States in 2006-2010[J]. Neuro Oncol, 2013,15(SUP 2):ii1-ii56.
• [16]MALMSTROM A,GRONBERG B H,MAROSI C, et al.Temozolomide versus standard 6-week radiotherapy versus hypofractionated radiotherapy in patients older than 60years with glioblastoma:the Nordic randomized, phase 3trial[J]. Lancet Oncol, 2012,13(9):916-926.
• [17]MIN W J,LI,ZHANG Y H,et al. Role of the anti-glioma drug ATI3148 in the inhibition of Notch signaling pathway[J]. Gene, 2015,573(1):153-159.
• [18]COHEN A L,COLMAN H. Glioma biology and molecular markers[J]. Cancer Treat Res, 2015,163(5):15-30.
• [19]SZE C I,SU W P,CHIANG M F,et al. Assessing current therapeutic approaches to decode potential resistance mechanisms in glioblastomas[J]. Front Oncol, 2013,3(3):59.
• [20]马福超．survivin与肿瘤治疗的研究进展[J]．中国癌症防治杂志，2010,2(3):239-241.
• [21]叶星星，蔡志毅，刘池波．细胞周期蛋白G1、G2与恶性肿瘤的研究进展[J]．温州医学院院报，2011,41(3):299-301.
• [22]LI T,SONG T,NI I,et al. The p-ERK-p-c-Jun-cyclinD1pathway ie involved in proliferation of smooth muscle cells after exposure to cigarette smoke extract[J]. Biochem Biophys Res Commun, 2014,453(3):316-320.
• [23]XIAO B,SPENCER J,CLEMENTS A,et al. Crystal structure of the retinoblastoma tumor suppressor protein bound to E2F and the molecular basis of its regulation.[J]. Proc Natl Acad Sci USA, 2003,100(5):2363-2368.
• [24]ALAO J P. The regulation of Cyclin D1 degradation:roles in cancer development and the potential for therapeutic invention[J]. Mol Cancer, 2007,6(4):24-28.
• [25]KIM J K,DIEHL J A. Nuclear CyclinD1:An oncogenic driver in human cancer[J]. Cell Physiol, 2009,220(2):292-29.

文章评论(Comment)：