Effects of Bushen Huoxue Recipe combined with neural stem cell transplantation in rats with tinnitus

doi:10.3969/j.issn.2095-4344.2017.05.013

Abstract

Abstract:

BACKGROUND: Neural stem cells have multi-directional differentiation potential, self-sustaining and self-renewal capacity as well as have strong migration ability. Bushen Huoxue Recipe can reduce neuronal damage and promote nerve cell regeneration, to achieve neural function reconstruction. Underlying mechanisms of Bushen Huoxue Recipe combined with neural stem cell transplantation in rats with tinnitus induced by sodium salicylate are yet unclear.
OBJECTIVE: To investigate the effects of Bushen Huoxue Recipe combined with neural stem cell transplantation in rats with tinnitus induced by sodium salicylate.
METHODS: Sixty Sprague-Dawley rats were randomized into five groups (n=12): normal control group, tinnitus model group, Bushen Huoxue Recipe group, stem cell group and combined treatment group (Bushen Huoxue Recipe combined with neural stem cell transplantation). Animal models of tinnitus induced by sodium salicylate were made in all the groups except for the normal control group. Fifteen days after modeling, rats were given intragastric administration of Bushen Huoxue Recipe water decoction (3 mL, 2.592 g/mL) for consecutive 7 days in the Bushen Huoxue Recipe group, intravenous injection of neural stem cells (1 mL, 1.0×10⁹/L) in the stem cell group, or their combined treatment in the combined treatment group.
RESULTS AND CONCLUSION: Bushen Huoxue Recipe, neural stem cell transplantation and their combination all could effectively promote the recovery of drinking water inhibitory rate that was ranked as follows: combined treatment group < Bushen Huoxue Recipe group < stem cell group. Additionally, the above-mentioned methods could effectively reduce pathological damage of primary auditory cortex in the rat brain. Western blot results showed higher 5-serotonin levels in the Bushen Huoxue Recipe group and combined treatment group than in the stem cell group, which were identical to ELISA findings. Our experimental findings indicate that Bushen Huoxue Recipe combined with neural stem cell transplantation exhibit certain effects on tinnitus induced by sodium salicylate in rats, and the mechanism of action is associated with neuronal repair and expression of 5-serotonin at mRNA and protein levels.

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: Tinnitus, Drugs, Chinese Herbal, Tissue Engineering

CLC Number:

R394.2

Wang Rong-guo, Pi Li-hong, Chen Hong-yao, Zhang Hai-zhong. Effects of Bushen Huoxue Recipe combined with neural stem cell transplantation in rats with tinnitus[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(5): 730-735.

Figures/Tables 2

2.1 实验动物数量分析实验所用动物数量为60只，在实验过程中并未有脱落情况。 2.2 动物耳鸣情况如图1所示，各组大鼠训练后条件反射建立，其Rb值均小于0.2时，提示条件反射成功形成；各组大鼠在条件反射消减期即在适应“安静”期间，每只大鼠的Rb值也均恢复至0.5左右。补肾活血方组、神经干细胞组及联合组大鼠在条件反射消减期(7 d内)的反射抑制率Rb的均值相对于模型组均有不同程度降低(P < 0.05)，反映补肾活血方和/或神经干细胞对水杨酸钠造成耳鸣的症状有改善治疗作用，且补肾活血方组的Rb值恢复速度较神经干细胞组慢，且联合组Rb恢复速度慢于补肾活血方组及神经干细胞组，已与正常对照组接近，说明补肾活血方组效果较神经干细胞组佳，联合给药组效果最佳。 2.3 大脑初级听皮质病理形态苏木精-伊红染色结果显示，正常对照组大鼠的神经元细胞排列规则、紧密。模型组大鼠神经元细胞不均匀，部分出现皱缩，毛细血管弯曲变形，细胞膜呈空泡状；而补肾活血方组、神经干细胞组及联合组大鼠初级听皮质中神经元细胞均减轻程度有所不同，治疗后改善情况从强到弱为：联合组>补肾活血方组>补肾活血方组，且其中联合组形态接近正常对照组(图2)，说明该组治疗效果最佳，与动物耳鸣情况结果相符。 2.4 脑组织中5-羟色胺表达水平 Western blot结果显示，正常对照组大鼠脑组织5-羟色胺表达水平较高；与正常对照组相比，模型组大鼠脑组织5-羟色胺表达水平明显下降(P < 0.05)；与模型组相比，补肾活血方组及联合组大鼠脑组织5-羟色胺表达水平均有所上调(P < 0.05)，但单纯神经干细胞移植不改变脑组织5-羟色胺中表达水平(P ≥ 0.05)，且补肾活血方组与联合组大鼠脑组织5-羟色胺表达水平高于神经干细胞组，差异有显著性意义(P < 0.05)，联合组大鼠脑组织5-羟色胺表达水平与正常对照组接近(P > 0.05；图3)。"

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