中医超声药透电疗仪改善脑缺血大鼠炎症反应的实验研究

Experimental study on the improvement of inflammatory response in rats with cerebral ischemia by Chinese ultrasound drug permeation electrotherapy device

目的观察中医超声药透电疗仪对脑缺血大鼠炎症反应的影响,为中医超声药透电疗仪治疗脑缺血的临床应用提供实验依据。方法将72只SD大鼠按随机分组法分为假手术组12只和造模组60只,造模组采用线栓法制备大脑中动脉闭塞(MCAO)模型。将造模成功大鼠按随机数字表法分为模型组、中药片组、空白片+中医超声药透电疗仪组(以下简称“空白片+电疗组”)、中药片+中医超声药透电疗仪组(以下简称“中药片+电疗组”)、丁苯酞组,每组12只。连续给予相应干预7 d后,进行神经功能评分;TTC染色观察脑梗死体积,并计算脑梗死体积;HE染色观察各组大鼠皮质区细胞形态;采用ELISA法检测各组大鼠血清TNF-α、IL-1β水平;采用Western blot法检测各组大鼠梗死侧海马组织TLR4、髓样分化因子(MyD88)、NF-κBp65蛋白表达情况。结果与模型组比较,空白片+电疗组、中药片+电疗组、丁苯酞组神经功能评分均降低(P<0.05),脑梗死体积降低(P<0.05),血清TNF-α、IL-1β水平降低(P<0.05),海马组织TLR4[(0.42±0.07)、(0.31±0.07)、(0.19±0.04)比(0.68±0.14)]、MyD88[(0.39±0.12)、(0.30±0.07)、(0.23±0.11)比(0.67±0.10)]、NF-κBp65[(0.32±0.03)、(0.27±0.02)、(0.17±0.03)比(0.57±0.12)]蛋白表达降低(P<0.05)。结论中医超声药透电疗仪可抑制MCAO大鼠海马组织TLR4、MyD88、NF-κBp65蛋白表达,降低血清炎症因子TNF-α、IL-1β水平,从而发挥神经保护作用。

Objective To observe the effects of Traditional Chinese Medicine(TCM)ultrasound drug permeation electrotherapy device on the inflammatory response of rats with cerebral ischemia,and to provide an experimental basis for the clinical application of TCM ultrasound drug permeation electrotherapy device in the treatment of cerebral ischemia.Methods A total of 72 SD rats were randomly divided into sham-operation group(12 rats)and modeling group(60 rats).The middle cerebral artery occlusion(MCAO)model was prepared by thread embolism in the model group.The rats were divided into model group,Chinese medicine tablet group,blank tablet+TCM ultrasound drug permeation electrotherapy group(hereinafter referred to as"blank tablet+electrotherapy group"),Chinese medicine tablet+TCM ultrasound drug permeation electrotherapy group(hereinafter referred to as"Chinese medicine tablet+electrotherapy group")and butylphthalide group according to the random number table method,with 12 rats in each group.The corresponding treatment was given continuously for 7 days.The neurological function was scored using Longa method evaluation criteria;TTC staining was used to observe the infarct volume and calculate the percentage of infarct volume;HE staining was used to observe the cell morphology of cortical area in each group of rats;ELISA was used to detect the serum TNF-αand IL-1βlevels in each group of rats;TLR4,MyD88 and NF-κBp65 protein expressions in hippocampal tissue of each group of rats on the infarct side were detected by Western blot method.Results Compared with the model group,the neurological function scores of rats in the blank tablet+electrotherapy group,the herbal tablet+electrotherapy group,and the butylphthalein group significantly decreased(P<0.05),the percentage of cerebral infarct volume significantly decreased(P<0.05),the contents of serum TNF-αand IL-1βsignificantly decreased(P<0.05),and the expressions of TLR4(0.42±0.07,0.31±0.07,0.19±0.04 vs.0.68±0.14),MyD88(0.39±0.12,0.30±0.07,0.23±0.11 vs.0.67±0.10),NF-κBp65(0.32±0.03,0.27±0.02,0.17±0.03 vs.0.57±0.12)protein in hippocampal tissue significantly decreased(P<0.05).Conclusion The TCM ultrasound drug permeation electrotherapy device can inhibit TLR4,MyD88,NF-κBp65 protein expressions and reduce the release of serum inflammatory factors TNF-αand IL-1β,thus exerting cerebral ischemic protective effects.