Combined effects of neural stem cell transplantation and electroacupuncture on the behaviors of chronic stress-induced depression rats

doi:10.3969/j.issn.2095-4344.2017.01.025

Abstract

Abstract:

BACKGROUND: Both electroacupuncture and neural stem cell transplantation have certain therapeutic effects on chronic stress depression, but their synergistic effects remain unclear.
OBJECTIVE: To explore the effect of electroacupuncture intervention combined with neural stem cell transplantation via tail vein on the behaviors of chronic stress-induced depression rats.
METHODS: Fifteen of 63 rats were randomly selected as normal group, and the other 48 rats were fed individually and underwent chronic unpredictable mild stress stimulation to make rat models of depression. Except for three unsuccessful models, 45 successful models were randomly assigned into model (no intervention), neural stem cell (injection of 1 mL neural stem cell suspension (3×106) via the tail vein), or combined treatment group (injection of 1 mL neural stem cell suspension (3×10⁶) via the tail vein+1-hour electroacupuncture every other day for consecutive 1 week) (n=15/group). The behavior ability of rats was assessed through sucrose preference test and open-field test. The survival, distribution and apoptosis of neural stem cells were assessed by CM-Dil labeling and TUNEL assay, respectively. The expression of Caspase-3 and Bcl-2 in the hippocampal CA3 region was assessed by RT-PCR and western blot assay.
RESULTS AND CONCLUSION: Compared with the normal group, model group rats exhibited lower body mass, lower scores on the sucrose preference test and open-field test, higher Caspase-3 mRNA and protein expressions (P < 0.05), lower Bcl-2 protein and mRNA expressions (P < 0.05) and more apoptotic neurons (P < 0.05). Compared with the model group, rats in the neural stem cell group and combined treatment group had significant increases in the body mass and scores on the sucrose preference test and open-field test as well as exhibited significantly decreased Caspase-3 mRNA and protein expressions (P < 0.05) and increased Bcl-2 protein and mRNA expressions (P < 0.05). These indicators were improved better in the combined treatment group than the neural stem cell group (P < 0.05). Under the fluorescence microscope, there were more CM-Dil-positive cells in the combined treatment group than the neural stem cell group. To conclude, neural stem cell transplantation can improve depressive behaviors in rats with chronic stress-induced depression, and electroacupuncture can elevate the survival rate of transplanted neural stem cells by reducing Caspase-3 and increasing Bcl-2 expression.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Depression, Electroacupuncture, Neural Stem Cells, Tissue Engineering

CLC Number:

R394.2

Mi Kun, Guo Qiang, Sun Zhen-qing, Wang Hai-bo. Combined effects of neural stem cell transplantation and electroacupuncture on the behaviors of chronic stress-induced depression rats[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(1): 140-145.

Figures/Tables 2

2.1 实验动物数量分析 63只雄性SPF级SD大鼠随机取15只不予任何处理为正常组，余48只应用孤养联合慢性不可预见性温和应激方法建立大鼠抑郁模型，剔除造模不成功的大鼠3只，将造模成功的45只大鼠随机分为3组，进入结果分析60只，中途无脱落。 2.2 大鼠行为学改变与正常组比较，模型组大鼠体质量、蔗糖水嗜好水平、旷场实验得分偏低(P < 0.05)。与模型组比较，神经干细胞组和电针刺激联合神经干细胞组大鼠体质量、蔗糖水嗜好水平、旷场实验得分明显增加(P < 0.05)，见表1-3。 2.3 CM-Dil标记的神经干细胞的存活和分布情况神经干细胞组海马CA3区可见CM-Dil阳性细胞，电针刺激联合神经干细胞组可见到较多CM-Dil阳性细胞，见图1。 2.4 TUNEL法测定神经细胞凋亡情况与正常组比较，模型组能见到更多神经细胞凋亡。与模型组比较，神经干细胞组神经细胞凋亡明显减少，电针刺激联合神经干细胞组改善更为突出，见图2。 2.5 RT-PCR检测大鼠海马CA3区Caspase-3、Bcl-2 mRNA表达与正常组比较，模型组大鼠海马CA3区Caspase-3基因表达显著升高(P < 0.05)；Bcl-2基因表达显著降低(P < 0.05)。与模型组相比，神经干细胞组和电针刺激联合神经干细胞组大鼠海马CA3区Caspase-3基因表达显著降低 (P < 0.05)，Bcl-2基因表达显著升高(P < 0.05)。电针刺激联合神经干细胞组各指标改善较神经干细胞组更明显(P < 0.05)，见图3。 2.6 Western blot检测大鼠海马CA3区Caspase-3、Bcl-2蛋白表达与正常组比较，模型组大鼠海马CA3区Caspase-3蛋白表达显著升高(P < 0.05)；Bcl-2蛋白表达显著降低(P < 0.05)。与模型组相比，神经干细胞组和电针刺激联合神经干细胞组大鼠海马CA3区Caspase-3蛋白表达显著降低 (P < 0.05)，Bcl-2蛋白表达显著升高(P < 0.05)。电针刺激联合神经干细胞组各指标改善较神经干细胞组更明显(P < 0.05)，见图4。"

References

[1] 黄洁云.抑郁症的发病机制与治疗进展[J].中国疗养医学,2013, 22(3):233-235.

[2] 毛榕榕,田孟,徐林.抑郁的大脑：抑郁症的神经生物学研究和抗抑郁新药研发[J].自然杂志,2009,31(3):148-152.

[3] Yu S, Holsboer F, Almeida OF. Neuronal actions of glucocorticoids: focus on depression. J Steroid Biochem Mol Biol. 2008;108(3-5):300-309.

[4] Duman RS, Kehne JH. Depression.CNS Neurol Disord Drug Targets. 2007;6(3):161-162.

[5] 胡随瑜,王素娥,张春虎.抑郁症临床症状分层与中医辨证分型的关系[J].中西医结合学报,2011,9(9):933-936.

[6] Han Y, Khodr CE, Sapru MK, et al. A microRNA embedded AAV α-synuclein gene silencing vector for dopaminergic neurons.Brain Res. 2011;1386:15-24.

[7] Kupfer DJ, Frank E, Phillips ML. Major depressive disorder: new clinical, neurobiological, and treatment perspectives. Lancet. 2012;379(9820):1045-1055.

[8] Alexopoulos GS, Murphy CF, Gunning-Dixon FM, et al. Serotonin transporter polymorphisms, microstructural white matter abnormalities and remission of geriatric depression. J Affect Disord. 2009;119(1-3):132-141.

[9] 崔青,孙丽亚,肖红,等.抑郁症患者神经内分泌节律变化的对照研究[J].中华行为医学与脑科学杂志,2010,19(1):33-35.

[10] 胡大吉,丁桂霞,李有田.细胞因子异常与抑郁症的发生[J].中华中医药学刊,2007,25(3):516-517.

[11] 张园,刘学军.海马神经元凋亡与抑郁症[J].国际精神学杂志, 2014,41(3):153-155.

[12] Drevets WC, Price JL, Furey ML. Brain structural and functional abnormalities in mood disorders: implications for neurocircuitry models of depression. Brain Struct Funct. 2008; 213(1-2):93-118.

[13] Dygalo NN, Kalinina TS, Bulygina VV, et al. Increased expression of the anti-apoptotic protein Bcl-xL in the brain is associated with resilience to stress-induced depression-like behavior. Cell Mol Neurobiol. 2012;32(5):767-776.

[14] Shishkina GT, Kalinina TS, Berezova IV, et al. Stress-induced activation of the brainstem Bcl-xL gene expression in rats treated with fluoxetine: correlations with serotonin metabolism and depressive-like behavior. Neuropharmacology. 2012; 62(1):177-183.

[15] Kubera M, Obuchowicz E, Goehler L, et al. In animal models, psychosocial stress-induced (neuro)inflammation, apoptosis and reduced neurogenesis are associated to the onset of depression. Prog Neuropsychopharmacol Biol Psychiatry. 2011;35(3):744-759.

[16] 户丽,梁佳,金树英,等.针刺治疗抑郁症作用机制近5年研究进展[J].针刺研究,2013,38(3):253-258.

[17] 孙丽荚,武燕峰.中医药治疗抑郁症研究进展[J].中医药报,2011, 39(6):107-108.

[18] 戴巍,李卫东,卢峻,等.电针对慢性应激抑郁大鼠海马神经元凋亡及JNK信号转导通路的影响[J].针刺研究,2010,35(5):330-334.

[19] 梁佳,李卫东,吴元坪,等.电针对慢性应激抑郁模型大鼠海马神经元凋亡与再生的影响[J].中华中医药杂志,2012,27(4): 947-950.

[20] 卢金花,姜劲峰,王玲玲,等.电针治疗抑郁模型大鼠快速起效海马区BDNF-TrkB机制研究[J].针灸临床杂志,2011,27(5):51-54.

[21] 孙健,刘健华,王谦,等.电针对抑郁症大鼠海马组织AC-cAMP- PKA信号通路的影响[J].中国老年学杂志,2010,30(24): 3672-3674.

[22] 刁波,刘琴,王丽萍,张宜.叶酸联合成体神经干细胞治疗创伤性脑损伤大鼠的实验[J].中国比较医学杂志,2012,22(7):48-55.

[23] 金树英,户丽,包伍叶,等.针刺对慢性应激抑郁大鼠血清和脑炎性细胞因子的影响[J].针灸临床杂志,2014,30(5):57-60.

[24] 乌吉斯古冷,佟海英,斯日古楞,等. 槟榔十三味丸(高尤-13)对慢性应激抑郁大鼠行为学及海马神经元凋亡的影响[J].中医学报, 2014,29(6):759-762.

[25] 张露芬.实验针灸学[M].北京:化学工业出版社,2010.

[26] 刘家言,万丹丹.人参皂苷对慢性应激抑郁模型小鼠的抗抑郁样作用[J].医药前言,2015,5(26):111-112.

[27] 程瑞凤,华冰,景晶,等.甘草总黄酮抗大鼠应激抑郁行为作用及对海马脑区神经细胞凋亡调控相关蛋白表达的影响[J].中药药理与临床,2014,30(2):69-72.

[28] 董爱国,赵丽丽,李海红.BDNF在抑郁症发病机制中的作用及其抗抑郁作用机制的研究进展[J].中国医疗前沿, 2011, 6(6):16-18.

[29] 包伍叶,张敏,杨秀岩,等.电针对慢性应激抑郁模型大鼠海马神经干细胞巢蛋白的影响[J].中国基层医药.2014,21(4):481-483.

[30] Kosten TA, Galloway MP, Duman RS, et al. Repeated unpredictable stress and antidepressants differentially regulate expression of the bcl-2 family of apoptotic genes in rat cortical, hippocampal, and limbic brain structures. Neuropsychopharmacology. 2008;33(7):1545-1558.

[31] Bachis A, Cruz MI, Nosheny RL, et al. Chronic unpredictable stress promotes neuronal apoptosis in the cerebral cortex. Neurosci Lett. 2008;442(2):104-108.

[32] van Breukelen F, Krumschnabel G, Podrabsky JE.Vertebrate cell death in energy-limited conditions and how to avoid it: what we might learn from mammalian hibernators and other stress-tolerant vertebrates. Apoptosis. 2010;15(3):386-399.

[33] 赵珂,金季玲.滋水清肝饮对围绝经期抑郁症模型大鼠海马神经元细胞凋亡相关因子的影响[J].环球中医药,2010,3(5):336-338.

[34] 黄泉智,许成勇,孙志高,等.舒郁宁心汤对抑郁大鼠海马Bcl-2和Bax表达的影响[J].中国康复理论与实践,2012,18(3):236-238.