补肾活血方联用神经干细胞移植治疗大鼠耳鸣

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

背景:神经干细胞具有多向分化的潜能及自我维持和更新能力,其迁徙能力强大,补肾活血中药可减轻神经元损伤及促进神经细胞再生修复,实现神经功能重建,补肾活血方联合神经干细胞应用于水杨酸钠耳鸣大鼠模型的作用机制尚不清楚。目的:探讨补肾活血方联合神经干细胞对耳鸣大鼠模型的影响。方法:取SD大鼠60只随机等分为5组,正常对照组、模型组、补肾活血方组、神经干细胞组和联合组,后4组建立水杨酸钠耳鸣大鼠模型,造模15 d后,补肾活血方单组每天灌胃3 mL灌胃补肾活血方水煎液(2.592 g/mL),连续7 d,神经干细胞组尾静脉注射1 m L神经干细胞培养液(1.0×109 L-1);联合组则先通过混悬液灌胃给药后再进行尾静脉注射神经干细胞培养液。结果与结论:(1)补肾活血方和/或神经干细胞治疗均能有效促进大鼠饮水抑制率的恢复,且饮水抑制率:联合组<补肾活血方组<神经干细胞组;(2)补肾活血方和/或神经干细胞治疗均能有效减轻大脑初级听皮质的病理损伤;(3)Western blot结果表补肾活血方组及联合组大鼠大鼠5-羟色胺表达水平较模型组增加,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;(4)ELISA法检测结果与Western blot结果一致;(5)表明补肾活血方联合神经干细胞对水杨酸钠耳鸣大鼠模型有一定疗效,其作用机制可能与修复神经元损伤及5-羟色胺含量及其蛋白表达有关。

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 m L, 2.592 g/m L） for consecutive 7 days in the Bushen Huoxue Recipe group, intravenous injection of neural stem cells（1 m L, 1.0×10^9/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 m RNA and protein levels.