棘脑通的抗抑郁作用及机制研究

Antidepressant Effects and Mechanism of Jinaotong

目的:考察主成分为沙棘和花生壳提取物的棘脑通的抗抑郁药效及作用机制。方法:(1)动物水平上,采用慢性不可预知温和应激(CUMS)抑郁模型小鼠,随机分为空白组、模型组、棘脑通给药组(棘脑通低浓度组、棘脑通高浓度组)和氟西汀组。通过体重测试、糖水偏嗜实验和悬尾不动时间测试等行为学研究,考察棘脑通的抗抑郁作用。(2)细胞水平上,将不同浓度的棘脑通应用于PC12细胞、SH-SY5Y细胞、BV2细胞和RAW 264.7细胞,研究棘脑通对神经营养因子功能的促进作用,保护神经细胞、诱导神经分化的作用,以及抗氧化、抗炎作用。结果:(1)与空白组相比,模型组小鼠糖水偏嗜率、体重显著降低,悬尾不动时间、强迫游泳不动时间显著延长(P<0.01);棘脑通给药组小鼠糖水偏嗜率较模型组显著升高(P<0.05),棘脑通给药组、氟西汀组小鼠体重较模型组显著升高(P<0.01),棘脑通低、高浓度组小鼠悬尾不动时间较模型组显著缩短(P<0.01、P<0.05),棘脑通给药组小鼠强迫游泳不动时间较模型组显著缩短(P<0.01),上述差异均有统计学意义。(2)棘脑通与神经生长因子(5、50 ng/mL)协同使用时,PC12细胞分化程度、pNF200-Luc表达荧光素酶活性显著提高。棘脑通与脑源性神经营养因子、神经营养因子-3和神经营养因子-4(5 ng/mL)协同使用时,荧光素酶的活性显著提高。100~1 000μg/mL的棘脑通可显著降低β-淀粉样蛋白的聚集。采用叔丁基过氧化氢诱导PC12细胞的氧化损伤后,应用棘脑通可提高PC12细胞活性,且浓度越高,效果越好。采用Erastin诱导SH-SY5Y细胞铁死亡后,应用棘脑通可提高SH-SY5Y细胞活性,且浓度越高,效果越好。将脂多糖用于BV2、RAW 264.7细胞后,应用棘脑通可降低白细胞介素1β的mRNA水平、核因子κB转录活性,且具有浓度依赖性。结论:棘脑通具有抗炎、抗氧化,保护神经细胞,诱导神经微丝表达,增强神经营养因子样的作用;该药通过以上作用发挥抗抑郁效果。

OBJECTIVE:To investigate the antidepressant effects and mechanism of Jinaotong(JNT) with extracts of sea buckthorn and peanut hull as its main components.METHODS:(1) At the animal level,chronic unpredictable mild stress(CUMS) depression model mice were randomly divided into the blank group,model group,JNT administration group(JNT low concentration group and JNT high concentration group) and fluoxetine group.The antidepressant effects of JNT were inspected by behavioral studies such as body weight test,sugar-water preference test and tail suspension immobility time test.(2) At the cellular level,different concentrations of JNT were applied to PC12 cells,SH-SY5Y cells,BV2 cells and RAW 264.7 cells,so as to probe into the promotional effects of JNT on the function of neurotrophic factors,the protection of neuronal cells and the induction of neural differentiation,as well as the antioxidant and anti-inflammatory effects.RESULTS:(1) Compared with the blank group,the sugar-water preference rate and body weight of the model group decreased significantly,the tail suspension immobility time and forced swimming immobility time of the model group were significantly prolonged(P<0.01).The sugar-water preference rate of JNT administration group was significantly higher than that of the model group(P<0.05),the body weight of the JNT administration group and fluoxetine group were significantly higher than that of the model group(P<0.01),the tail suspension immobility times of the JNT low concentration group and JNT high concentration group were significantly shorter than that of the model group(P<0.01,P<0.05),the forced swimming immobility time of the JNT administration group was significantly shorter than those of the model group(P<0.01),the differences were statistically significant.(2) When JNT was synergistically administered with nerve growth factor(5 and 50 ng/mL),the differentiated degree of PC12 cell and activity of pNF200-Luc expression luciferase increased significantly.The luciferase activity increased significantly when JNT was synergistically administered with brain-derived neurotrophic factor,neurotrophin-3 and neurotrophin-4(5 ng/mL).Dose from 100 to 1 000 μg/mL of JNT could significantly reduced amyloid β-protein aggregation.After the oxidative damage of PC12 cells induced by tBHP,the application of JNT can increased the activity of PC12 cells,the higher the concentration,the better the effect.After the iron death of SH-SY5Y cells induced by Erastin,the application of JNT can increase the activity of SH-SY5Y cells,the higher the concentration,the better the effect.After using lipopolysaccharide on BV2 and RAW 264.7 cells,the application of JNT can reduce the mRNA level of interleukin-1β and the nuclear factor kappa B transcriptional activity,which showed a concentration-dependent manner.CONCLUSIONS:JNT has the effects of anti-inflammatory,antioxidant,protecting nerve cells,inducing the expression of neurofilaments,and enhancing the neurotrophic factor-like;the JNT exerts antidepressant effects through the above effects.