针刺对急性颅内出血大鼠的神经功能及损伤的影响

Effect of the acupuncture and moxibustion in the neurological function and treatment of injury in rats with acute intracranial hemorrhage

目的观察针刺对急性颅内出血大鼠神经功能状态及对颅脑损伤情况的改善效果。方法选用6~8周约220 g健康的雄性SD大鼠共45只,采用随机数字表法将其分为假手术组、模型组和针刺组,各15只,每组再分为1、3和7 d的3个时间点,各5只。用自体血注射的方式而构建脑出血大鼠模型组。假手术组只钻孔不注射自体血,针刺组采用由百会穴刺向曲鬓穴的治疗方法。采用Ludmila Belayev神经功能评分评估各组大鼠神经功能系统的缺损程度。采用Western blot检测脑组织中CTRP3、GPX4蛋白的表达。用透射电镜观测各组大鼠脑组织的神经细胞线粒体形态。qRT-PCR法检测大鼠脑组织中的GFAP和EGFR mRNA特异性表达。结果与假手术组比较,模型组各时间点神经功能缺损评分均增加(P<0.05)。与模型组比较,针刺组神经功能缺损评分1 d和3 d降低,差异有统计学意义(P<0.05),在7 d时系统功能缺损评分比较,差异无统计学意义(P>0.05)。与假手术组比较,模型组各时间点CTRP3和GPX4蛋白表达均降低,差异有统计学意义(P<0.05);与模型组比较,针刺组各时间点CTRP3和GPX4蛋白表达水平上调,差异有统计学意义(P<0.05)。与假手术组比较,模型组各时间点大鼠脑组织GFAP、EGFR mRNA水平增加,差异有统计学意义(P<0.05)。与模型组比较,针刺组各时间点大鼠脑组织GFAP、EGFR mRNA水平均明显降低,差异有统计学意义(P<0.05)。结论针刺治疗可以上调脑组织中CTRP3和GPX4蛋白表达,降低GFAP、EGFR的mRNA水平,同时恢复神经元线粒体的完整性,从而改善急性脑出血大鼠神经肌肉损伤神经功能,在脑出血治疗中发挥保护作用。

Objective To observe the improvement effect of acupuncture on the neurological function and craniocerebral injury in rats with acute intracranial hemorrhage.Methods A total of 45 healthy male SD rats of about 220 g in 6-8 weeks were selected,and they were divided into sham operation group,model group and acupuncture group by random number table method,with 15 rats in each group;each group was divided into three points at 1,3 d and 7 d,with 5 rats in each time.The cerebral hemorrhage rat model group was established by autologous blood injection;the sham operation group only drilled holes without injecting autologous blood;the acupuncture group adopted the treatment method of piercing from Baihui to Qubin.Ludmila Belayev’s neurological function score was used to evaluate the degree of neurological deficit in rats in each group.Western blot was used to detect the expression of CTRP3 and GPX4 proteins in brain tissue.The mitochondrial morphology of nerve cells in the brain tissue of rats in each group was observed by transmission electron microscope.qRT-PCR method was used to detect the specific expression of GFAP and EGFR mRNA in rat brain tissue.Results Compared with sham operation group,neurological function deficit score in model group increased at each time point(P<0.05).Compared with model group,neurological impairment scores of acupuncture group decreased at 1 and 3 d,the differences were statistically significant(P<0.05),but there was no statistically significant difference in systemic impairment score at 7 d(P>0.05).Compared with sham operation group,levels of CTRP3 and GPX4 protein in model group were decreased at each time point,and the differences were statistically significant(P<0.05).Compared with model group,CTRP3 and GPX4 protein expression levels in acupuncture group were up-regulated at each time point,and the differences were statistically significant(P<0.05).Compared with sham operation group,mRNA levels of GFAP and EGFR in brain tissue of rats in model group increased at each time point,and the differences were statistically significant(P<0.05).Compared with model group,mRNA levels of GFAP and EGFR in brain tissue of rats in acupuncture group were significantly decreased at each time point,and the differences were statistically significant(P<0.05).Conclusion Acupuncture treatment can up-regulate the expression of CTRP3 and GPX4 proteins in brain tissue,reduce the mRNA levels of GFAP and EGFR,and restore the integrity of neuronal mitochondria,thereby improving the neuromuscular injury and neurological function of rats with acute cerebral hemorrhage,and play a protective role.