夹脊电针对急性脊髓损伤大鼠脊髓病理形态及组织中炎性因子表达的影响

目的:观察夹脊电针对急性脊髓损伤(ASCI)大鼠的肢体运动功能、脊髓组织病理形态学以及组织中IL-18、IL-1β和cleaved caspase-1表达的影响。方法:选用36只雌性SD大鼠,所有样本随机均分为假手术组(Sham组)12只、模型组(Model组)12只及夹脊电针组(EA组)12只,共3组。每组大鼠再按两个时间点分为3天组与7天组。夹脊电针组每日给予针刺治疗。治疗结束后,各组大鼠运动功能通过BBB评分法测定;经HE染色法观察各组大鼠脊髓组织病理形态学变化;ELISA法检测大鼠脊髓组织IL-18、IL-1β表达;免疫组化法检测脊髓组织中cleaved caspase-1的表达。结果:Sham组大鼠未见明显运动功能障碍,而Model组大鼠造模后运动功能障碍明显,3天、7天BBB评分均明显降低,与Sham组比较差异具有统计学意义(P<0.05)。EA组大鼠术后BBB评分,在3天、7天较术前升高,与模型组比较差异具有统计学意义(P<0.05)。Sham组大鼠脊髓组织形态正常,而Model组3天时大鼠脊髓组织可见明显的病理学改变,大量炎性细胞浸润,炎性反应明显,3天、7天时脊髓组织炎性细胞浸润及胶质细胞增生。夹脊电针治疗后可见组织结构趋于完整,炎性细胞及胶质细胞减少。Sham组大鼠脊髓组织可以明显看出有一小部分IL-18、IL-1β表达,在3天、7天节点中处于稳定状态。在3天、7天节点中,Model组大鼠脊髓组织IL-18、IL-1β表达出现升高情况,相较于Sham组情况有明显差异,且差异具有统计学意义(P<0.05)。经过治疗后,EA组大鼠脊髓组织中IL-18、IL-1β的表达均较治疗前减少,在7天节点中与Model组比较,差异具有统计学意义(P<0.05)。另外,Sham组大鼠脊髓组织中可以见到少量的cleaved caspase-1呈阳性表达,在3天、7天节点表达情况稳定。在3天、7天节点中Model组大鼠脊髓组织cleaved caspase-1阳性细胞的平均光密度值有明显的变化,且呈现升高的趋势,与Sham组比较差异有统计学意义(P<0.05)。EA组大鼠在经针刺治疗后,其脊髓组织中cleaved caspase-1阳性细胞的平均光密度值明显降低,与Model组比较差异具有统计学意义(P<0.05)。结论:夹脊电针能够通过抑制脊髓损伤大鼠脊髓组织中IL-18、IL-1β、cleaved caspase-1的表达,从而减轻炎性反应,促进运动功能恢复。

Effect of Jiaji Electro-Acupuncture on Pathological Changes and Its Influences to Expressions of Inflammatory Factors in Rats with ASCI

Objective:To observe the effect of Jiaji electro-acupuncture on limb movement function,pathological morphology of spinal cord tissues,and the expressions of IL-18,IL-1βand cleaved caspase-1 in spinal cord tissues in rats with acute spinal cord injury(ASCI).Methods:36 female SD rats were randomly divided into the sham-operation group(n=12),the model group(n=12)and the electro-acupuncture group(n=12).The rats in each group were further divided into the 3d group and the 7d group according to the postoperative time points.Acupuncture treatment was given daily in the Jiaji electro-acupuncture group.After the treatment,the motor function of each group was measured by BBB scores.The pathological morphology changes of spinal cord tissues in rats were observed by HE staining method.The expressions of IL-18 and IL-1βin spinal cord tissues were detected by ELISA method,and the expression of cleaved caspase-1 in spinal cord tissues was detected by immuno-histochemistry method.Results:There was no significant motor dysfunction in the sham-operation group;whereas in the model group,the motor dysfunction was significant,and BBB scores on 3rd day and on 7th day were significantly decreased compared to those in the sham-operation group(P<0.05).BBB scores were significantly increased in the electro-acupuncture group than those in the model group of rats in the jiaji electro-acupuncture group increased on 3rd day and on 7th day(P<0.05).There was no obvious morphology changes of spinal cord tissues in the sham-operation group;whereas significant pathological changes and obvious inflammatory responses,such as a mounts of inflammatory cells infiltration and gliocyte proliferation,were observed in the model group.After electro-acupuncture intervention,the tissue structure of spinal cord tended to be intact and the inflammatory cells and glial cells were decreased.A small amount of IL-18 and IL-1βcan be clearly seen in the spinal cord tissues in the sham-operation group,which showed a stable state on 3rd day and on 7th day.The expressions of IL-18 and IL-1βwere increased in the model group on 3rd day and on 7th day compared to those in the sham-operation group(P<0.05).After the electro-acupuncture treatment,the expressions of IL-18 and IL-1βwere decreased compared to those before the treatment;on 7th day,of which there were statistical differences between the model group and the electro-acupuncture group(P<0.05).In addition,a small number of cleaved caspase-1 expression was found in the spinal cord tissues in the sham-operation group,and the expression was stable on 3rd day and on 7th day.The average optical density value of cleaved caspase-1 positive cells was significantly increased in the model group on 3rd day and on 7th day compared to those in the sham-operation group(P<0.05).After electro-acupuncture treatment,the average optical density of cleaved caspase-1 positive cells was significantly decreased compared to those in the model group(P<0.05).Conclusion:Jiaji electro-acupuncture can reduce inflammatory response and promote the recovery of motor function by inhibiting the expressions of IL-18,IL-1βand cleaved caspase-1 in spinal cord tissues of rats with ASCI.