泡泡浴水疗对新生期大鼠反复惊厥性脑损伤保护作用的探讨

目的 探讨泡泡浴水疗对新生期大鼠反复惊厥性脑损伤是否具有保护作用.方法 出生6d(P6)的健康SD大鼠随机分为4组各15只,惊厥组(R5组)和惊厥水疗组(HRS组)在P6吸入三氟乙醚诱导惊厥发作,持续30 min,连续6d,对照组(CON组)和对照水疗组(HCON组)同样操作但不吸入三氟乙醚.于末次惊厥后第2天对HCON及HRS组采用泡泡浴进行干预,连续28d.观测各组大鼠体格发育及神经行为表现,P50时通过免疫印迹技术检测大脑皮质可塑性相关基因-1(Plasticity related gene 1,PRG-1)、抗凋亡基因Bc1-2蛋白的表达.结果 1.惊厥后RS组大鼠各日龄体质量较CON组显著降低(P＜0.05)；水疗后HRS组体质量较RS组显著增高(P＜0.05).各组生理发育现象(竖耳、门齿萌出、睁眼)出现时间无显著差异.2.旷场行为实验:RS组旷场得分较CON组明显降低(P＜0.05),HRS组旷场得分较RS组明显升高(P＜0.05).泡泡浴可增强反复惊厥大鼠的兴奋性和探索能力 P26开场得分:CON组为(45.78±18.31)分,HCON组为(59.78 ±25.61)分,RS组为(27.78±16.83)分,HRS组为(53.15±22.43)分,P＜0.01].3.4组大鼠大脑皮质PRG-1、Bc1-2蛋白表达存在显著差异(F=3.477/7.36,P＜0.05).其中RS组PRG-1(1.149±0.191)表达升高、Bcl-2(1.040±0.116)表达降低,与CON组(PRG-1为0.910±0.162,Bcl-2为1.259±0.157)比较均差异有显著性(P＜0.05)；HRS组Bcl-2(1.572±0.333)表达水平较RS组明显升高(P＜0.05).结论 新生期大鼠反复惊厥导致生长发育延迟、神经行为损伤,惊厥后早期进行泡泡浴干预对神经损伤有修复作用,大脑皮质PRG-1、Bcl-2参与发育期惊厥性脑损伤病理生理机制.

Protective effects of bubble bath hydrotherapy on brain damage of neonatal rats with recurrent seizures

Objective To explore the potentiality of protection by bubble bath hydrotherapy to brain damage of neonatal rats with recurrent seizures.Methods Sixty Sprague-Dawley neonatal rats were randomly divided into control group ( CON),control hydrotherapy group ( HCON ),recurrent-seizure group (RS) and recurrent-seizure hydrotherapy group (HRS).Each group had 15 rats.The volatile agent flurothyl was used to induce 30 min seizure attack.At postnatal day 6 (P6),recurrent seizures were induced once per day for consecutive 6 days in the RS group and HRS group.Rats in the CON and HCON group were placed into the container for same time as their counterpart without exposure to flurothyl.In HCON and HRS groups,Bubble bath hydrotherapy intervention was used after the end of the last seizure for 28 consecutive days.Inspections were made for maturation of physical characteristics,neural behavior.PRG-1 and Bcl-2 protein level in cerebral cortex were detected by western blot method at P50.Results 1.Maturation of physical characteristics:body weights of the rats in the RS group were decreased significantly (P＜0.05).Body weights of HRS rats were significantly increased than that of RS rats(P＜0.05 ).There were no significant difference of physiological development phenomenon among the five groups (P ＞0.05 ).2.There was high nerve excitation in hydrotherapy group ( At P26,the scores of OFT respectively were 45.78 ± 18.31 for CON,59.78 ± 25.61 for HCON,27.78 ± 16.83 for RS and 53.15 ± 22.43 for HRS,P ＜ 0.01 ).3.The expression of PRG-1 in cerebral cortex of RS group were significantly higher than that of CON group (1.149 ±0.191 for RS,0.910 ±0.162 for CON,P＜ 0.05 ).The level of Bcl-2 in cerebral cortex of RS group ( 1.149 ±0.191 ) were significantly lower than that of CON group( 1.259 ±0.157) (P＜0.05 ),the level of Bcl-2in cerebral cortex of HRS group( 1.572 ± 0.333 ) were significantly higher than that of RS group(P＜ 0.05 ).Conclusion Maturation of physical characteristics and neurobehavioral were damages following recurrent neonatal seizures,which may be associated with significantly increased level of PRG-1 protein and decreased level of Bcl-2protein in cerebral cortex.The early intervention by Bubble bath hydrotherapy can play a positive role on developmental rehabilitation.