乙酰葛根素对Aβ25-35诱导BV-2小胶质细胞Caspase-3表达的影响

doi:10.6040/j.issn.1671-7554.0.2015.428

摘要/Abstract

摘要： 目的探究乙酰葛根素对β淀粉样蛋白(Aβ_25-35)诱导BV-2小胶质细胞形态及含半胱氨酸的天冬氨酸蛋白水解酶-3(Caspase-3)表达的影响。方法用凝聚态Aβ_25-35诱导体外培养的BV-2小胶质细胞作为阿尔茨海默病炎症细胞模型,将细胞分为对照组、模型组、Caspase-3抑制剂(Z-DEVD-fmk)组以及乙酰葛根素低(0.1 μmol/L)、中(0.4 μmol/L)、高(1.6 μmol/L)剂量共6组,采用倒置相差显微镜观察小胶质细胞形态变化,应用Western blotting检测Caspase-3蛋白表达水平。结果形态学结果显示,Aβ_25-35可激活小胶质细胞使其由静息态变为阿米巴样,Caspase-3抑制剂、乙酰葛根素可改善Aβ_25-35诱导的小胶质细胞形态改变。Western blotting结果显示,与对照组相比,模型组Caspase-3表达显著升高,差异有统计学意义(P<0.01);与模型组比较,Caspase-3抑制剂、乙酰葛根素各剂量组Caspase-3表达均显著降低(P<0.01);与Caspase-3抑制剂组相比,乙酰葛根素低、中剂量组Caspase-3表达均显著升高(P<0.01),高剂量组则无统计学意义。结论乙酰葛根素可抑制Aβ诱导BV-2小胶质细胞激活,其机制可能与降低Caspase-3表达有关,且以高剂量组效果较好。

关键词: Caspase-3, 小胶质细胞, 乙酰葛根素, β淀粉样蛋白, Western blotting, 阿尔茨海默病

Abstract: Objective To investigate the effect of acetylpuerarin on the expression of Caspase-3 in BV-2 microglia induced by Aβ_25-35. Methods BV-2 microglia cells were activated with condensed Aβ_25-35 as Alzheimer's disease inflammatory cell model, which were then divided into 6 groups: control group, model group, Caspase-3 inhibitor (Z-DEVD-fmk) group and acetylpuerarin groups (low-dose group 0.1 μmol/L, moderate-dose group 0.4 μmol/L, high-dose group 1.6 μmol/L). Morphological changes of microglia cells were observed with an inverted phase contrast microscope, and the expression level of Caspase-3 protein in each group was detected by Western blotting. Results It was observed that Aβ_25-35 activated microglia from resting state into ameboid, and both Caspase-3 inhibitor and acetylpuerarin improved cell morphological changes induced by Aβ_25-35. Western blotting results showed that compared with the control group, the model group had significantly increased expression of Caspase-3 (P<0.01); compared with the model group, Caspase-3 inhibitor group and acetylpuerarin groups had markedly reduced expression of Caspase-3 (P<0.01); compared with Caspase-3 inhibitor group, acetylpuerarin low-dose and moderate-dose group had significantly increased expression of Caspase-3 (P<0.01), while acetylpuerarin high-dose group had no statistically significant difference. Conclusion Acetylpuerarin can inhibit activation of BV-2 microglia cells induced by Aβ_25-35. The mechanism may be related to decreased expression of Caspase-3, especially high-dose acetylpuerarin.

Key words: Alzheimer's disease, Acetylpuerarin, Caspase-3, Microglia, Western blotting, β-amyloid protein

中图分类号:

R741

李梅, 孟庆慧, 蔡巧英, 徐雁, 范晓婷. 乙酰葛根素对Aβ_25-35诱导BV-2小胶质细胞Caspase-3表达的影响[J]. 山东大学学报（医学版）, 2015, 53(10): 32-36.

LI Mei, MENG Qinghui, CAI Qiaoying, XU Yan, FAN Xiaoting. Effect of acetylpuerarin on the expression of caspase-3 in BV-2 microglia induced by Aβ_25-35[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2015, 53(10): 32-36.

参考文献

[1] Capiralla H, Vingtdeux V, Zhao H, et al. Resveratrol mitigates lipopolysaccharide- and Aβ-mediated microglial inflammation by inhibiting the TLR4/NF-κB/STAT signaling cascade[J]. J Neurochem, 2012, 120(3):461-472.
[2] Solito E, Sastre M. Microglia function in Alzheimer's disease [J]. Front Pharmacol, 2012, 3: 14. doi: 10.3389/fphar.2012.00014.
[3] 刘晓丽, 孟庆慧, 张晓青. 乙酰葛根素对Aβ_25-35诱导的BV-2小胶质细胞活性的影响[J].潍坊医学院学报, 2014, 36(3): 166-169. LIU Xiaoli, MENG Qinghui, ZHANG Xiaoqing. Effect of acetylpuerarin on Aβ_25-35-induced activation of BV-2 microglia[J]. Acta Acad Med Weifang, 2014, 36(3):166-169.
[4] Burguillos MA, Deierborg T, Kavanagh E, et al. Caspase signalling controls microglia activation and neurotoxicity[J]. Nature, 2011, 472(7343):319-324.
[5] 毛翘, 崔春爱. 小胶质细胞和神经炎症病变与阿尔茨海默病的相关性研究进展[J]. 延边大学医学院学报, 2012, 35(2):150-153.
[6] El Khoury J, Hickman SE, Thomas CA, et al. Scavenger receptor-mediated adhesion of microglia to beta-amyloid fibrils[J]. Nature, 1996, 382(6593):716-719.
[7] Roy A, Fung YK, Liu X, et al. Up-regulation of microglial CD11b expression by nitric oxide[J]. J Biol Chem, 2006, 281(21):14971-14980.
[8] 贾丽艳, 拓西平. 小胶质细胞与阿尔茨海默病[J]. 老年医学与保健, 2005, 4: 251-253. JIA Liyan, TUO Xiping. Microglia and Alzheimer disease[J]. Geriatr Health Care, 2005, 4:251-253.
[9] Harkany T, Abraham I, Konya C, et al. Mechanisms of beta-amyloid neurotoxicity: perspectives of pharmacotherapy[J]. Rev Neurosci, 2000, 11(4):329-382.
[10] Toro VC, Tehranian R, Zetterstrm M, et al. Increased gene expression of interleukin-1alpha and interleukin-6 in rat primary glial cells induced by beta-amyloid fragment[J]. J Mol Neurosci, 2001, 17(3):341-350.
[11] 矫健, 薛冰, 王晓民, 等. Aβ_25-35激活小胶质细胞机制的研究[J]. 中华神经医学杂志, 2007, 6(6):571-574. JIAO Jian, XUE Bing, WANG Xiaomin, et al. Research in mechanism of Aβ_25-35-activated microglias[J]. Chinese Journal of Neuromedicine, 2007, 6(6):571-574.
[12] 王灿茂, 程玉芳, 吴金刚, 等. 新型PDE4抑制剂Roflupram改善阿尔茨海默病大鼠的认知障碍及神经炎症[J]. 中国药理学通报, 2015, 31(3):327-333. WANG Canmao, CHENG Yufang, WU Jingang, et al. The new PDE4 inhibitor Roflupram ameliorated cognitive deficits and neuroinflammation in a rat model of Alzheimer's disease [J]. Chinese Pharmacological Bulletin, 2015, 31(3):327-333.
[13] 刘东梅, 张岫美, 娄凤兰. 乙酰葛根素对氧糖剥离诱导海马神经元凋亡的影响[J]. 中国药理学通报, 2013, 7:981-984. LIU Dongmei, ZhANG Xiumei, LOU Fenglan. Effect of Acetylpuerarin on apoptosis induced by oxygen-glucose deprivation/reperfusion in hippocampal neurons[J]. Chinese Pharmacological Bulletin, 2013, 7:981-984.
[14] Meng QH, Lou FL, Hou WX, et al. Acetylpuerarin reduces inflammation and improves memory function in a rat model of Alzheimer's disease induced by Abeta1-42[J]. Pharmazie, 2013, 68(11):904-908.
[15] 刘敏, 孟庆慧, 郭鹤, 等. 乙酰葛根素对阿尔茨海默病大鼠蛋白激酶-C-δ及白细胞介素-6表达的影响[J]. 解剖学报, 2013, 44(3):320-324. LIU Min, MENG Qinghui, GUO He, et al. Effects of Acetylpuerarin on the expression of protein kinase C-δ and interleukin-6 in Alzheimer disease rats[J]. Acta Anatomica Sinica, 2013, 44(3):320-324.
[16] 刘晓丽. 乙酰葛根素对Aβ诱导BV-2小胶质细胞炎性因子分泌的影响及机制研究[D].潍坊: 潍坊医学院, 2014.
[17] Gorman AM, Orrenius S, Ceccatelli S. Apoptosis in neuronal cells: role of caspases[J]. Neuroreport, 1998, 9(10): R49-55.
[18] Gervais FG, Xu D, Robertson GS, et al. Involvement of caspases in proteolytic cleavage of Alzheimer's amyloid-beta precursor protein and amyloidogenic A beta peptide formation[J]. Cell, 1999, 97(3):395-406.
[19] Takuma H, Tomiyama T, Kuida K, et al. Amyloid beta peptide-induced cerebral neuronal loss is mediated by caspase-3 in vivo[J]. J Neuropathol Exp Neurol, 2004, 63(3):255-261.
[20] 张晓杰, 董海影, 孙玉荣, 等. 大鼠大脑海马注射β-淀粉样蛋白对caspase-3蛋白表达的影响及通络救脑口服液的干预作用[J]. 中华中医药杂志, 2010, 10:1678-1680. ZhANG Xiaojie, DONG Haiying, SUN Yurong, et al. Expression of protein caspase-3 in Aβ_1-40 induced AD rats and effect of Tong luo Jiu'nao Oral Solution[J]. China Journal of Traditional Chinese Medicine and Pharmacy, 2010, 10:1678-1680.

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