回神颗粒减轻中度创伤性脑损伤大鼠脑水肿的疗效机制

目的 探讨回神颗粒对中度创伤性脑损伤（TBI）脑水肿的疗效机制。方法 应用液压性脑损伤装置建立中度TBI模型，创伤压力为（170±10） kPa，作用时间为（20±2） ms，随机将大鼠分为6组:正常组、假手术组、模型3 d组、模型7 d组、治疗3 d组、治疗7 d组。治疗3 d组和治疗7 d组于创伤后即刻用2 mL药液（含回神颗粒0.27 g）灌胃1次，此后再分别于每日早晚各灌胃1次，每次2 mL药液。采用双光束荧光分光光度计测定6组动物脑细胞内游离钙浓度；采用荧光定量PCR检测6组动物脑组织内N-甲基-D-天门冬氨酸受体1（NMDAR-1）、水通道蛋白4（AQP-4）的变化。采用光镜和扫描电镜观察6组动物脑损伤区脑细胞的病理形态学改变。结果 与正常组比较:模型3 d组细胞内游离钙明显升高，差异有统计学意义（P<0.01）。治疗3 d组细胞内游离钙的浓度较模型3 d组明显降低，差异具有统计学意义（P<0.01）。与正常组比较:模型3 d组、模型7 d组NMDAR-1及AQP-4的表达均明显升高，差异具有统计学意义（P<0.01）。治疗3天组NMDAR-1及AQP-4的表达均较模型3 d组明显降低，差异具有统计学意义（P<0.05）。光镜和电镜下观察模型3 d组、模型7 d组、治疗3 d组和治疗7 d组均可见到明显的脑细胞损伤的病理形态学改变。与正常组和假手术组比较，存在明显区别；与模型3 d组和模型7 d组比较，应用回神颗粒后，治疗3 d组和治疗7 d组的脑损伤区病理形态学改变明显减轻。结论 回神颗粒能够减轻创伤性脑水肿，维持细胞的正常结构，减轻创伤性脑损伤。其作用机制为可能为遏制钙超载、减轻NMDAR-1及AQP-4的过度表达。

The Mechanism Research of Huishen Granule Alleviate Hydrocephalus on Moderate Traumatic Brain Injury Model Rats

Objective To investigate the mechanism of Huishen Granule on hydrocephalus of moderate traumatic brain injury (TBI). Methods Using the equipment of fluid percussion injury (FPI) to establish moderate FPI model in SD rats. The traumatic pressure was (170±10) kPa, traumatic time was (20±2) ms. The rats were randomly divided into 6 groups:normal group, sham group, model 3 d group, model 7 d group, treatment 3d group, treatment 7 d group. The 3 d and 7 d treatment group were immediately given 2 mL fluid (contain 0.27 g Huishen Granule) by gavage. Then the rats were given by gavage twice for 2 mL every time in morning and evening. The concentration ofCa²⁺] in brain cells was examined by using double-beam fluorescent spectrophotometer. The change of NMDAR-1 and AQP-4 in all 6 groups was determined by using fluorescence quantitative polymerization chain reaction (PCR). The TBI pathological change of all 6 groups were observed by using optical microscope and election microscope. Results Compared with the normal group, theCa²⁺] of model 3 d group was increased obviously, the difference was statistically significant (P<0.01). Compared with model 3 d group, theCa²⁺] of treatment 3 d group was decreased obviously, with statistical significance (P<0.01). Compared with the normal group, the NMDAR-1 and AQP-4 expressions of model 3 d group and model 7 d group were increased obviously, which had statistical significance (P<0.01). Compared with model 3 d group, the NMDAR-1 and AQP-4 of treatment 3 d group descended significantly, which had statistic significance (P<0.05). The TBI pathological change of model 3 d group, model 7 d group, treatment 3 d group and treatment 7 d group were observed under optical microscope and election microscope. Compared with the normal and sham-operation groups, it had obvious difference. Compared with model 3 d group and model 7 d group were significantly alleviated. Conclusion Huishen Granule can alleviate traumatic hydrocephalus, maintain the normal structure of neurone, and lessen the TBI. The mechanisms probably were by inhibiting calcium overload and down regulating the NMDAR-1 and AQP-4 expressions.