摘要
目的:观察火针对神经细胞凋亡及其相关靶点关键蛋白表达的影响,探讨火针改善脊髓损伤(SCI)后神经细胞凋亡的机制。方法:将72只SD大鼠随机分为假手术组、模型组和火针组,每组分为1 d、3 d、7 d和14 d共4个时点。采用电动颅脑脊髓损伤撞击仪制备SCI模型,对火针组采用相应的干预方法,相应时点进行神经功能评分(BBB)、TUNEL检测和Western-blot检测。结果:BBB行为学评分显示,与假手术组比较,火针组在1 d、3 d、7 d及14 d神经运动功能情况均明显较低,差异具有统计学意义(P<0.05);与模型组比较,火针组在1 d、3 d、7 d及14 d神经运动功能情况均明显较高,差异具有统计学意义(P<0.05)。神经细胞凋亡结果显示,SCI后,与假手术组相比,模型组凋亡细胞明显增多,凋亡指数明显增加;与模型组相比,火针组大鼠脊髓损伤部位凋亡细胞减少,凋亡指数明显降低,差异具有统计学意义(P<0.05或P<0.01),且火针组凋亡指数14 d时最低。DAP12、p-PI3K及TGF-β蛋白表达结果显示,与假手术组相比,模型组大鼠DAP12蛋白相对表达量在各个时点均有增高的趋势,差异无统计学意义(P>0.05),与模型组比较,火针组在各个时点DAP12蛋白相对表达量有增高的趋势,7 d时增加最明显,差异无统计学意义(P>0.05);与假手术组比较,模型组p-PI3K蛋白相对表达量在各个时点均有增高的趋势,仅在1 d时差异有统计学意义(P<0.05),与模型组比较,火针组在1 d、3 d和14 d时p-PI3K蛋白相对表达量降低,在7 d时p-PI3K蛋白相对表达量明显升高,差异具有统计学意义(P<0.05);与假手术组比较,模型组在1 d、7 d与14 d时TGF-β蛋白表达量均增高,仅在14 d时差异有统计学意义(P<0.05),与模型组比较,火针组在3 d、7 d时TGF-β蛋白表达量高于模型组,但仅在3 d时差异具有统计学意义(P<0.01)。结论:火针能调控SCI后p-PI3K、TGF-β及DAP12蛋白的表达,从而改善神经细胞凋亡,促进SCI后神经运动功能恢复。
Objective:To observe the effect of fire needling on neuronal apoptosis and the expressions of key proteins of related targets,and to explore the mechanism of fire needling in improving neuronal apoptosis after spinal cord injury(SCI).Methods:72 SD rats were randomly divided into the sham operation group,the model group and the fire needling group,and each group was divided into 4 time points according to 1 d,3 d,7 d and 14 d.SCI model was prepared by electro-encephalitic impingement instrument for SCI model,and corresponding intervention method was applied in the fire needling group.The score of Basso Beattie Bresnahan(BBB)for neurological function,TUNEL staining and Western-blot detection were performed at corresponding time points.Results:The scores of BBB showed that compared with that in the sham operation group,the neuromotor function was decreased in the model group(P<0.05);compared with that in the model group,the neuromotor function was significantly higher in the fire needling group on 1 d,3 d,7 d and 14 d(P<0.05).The results of nerve cell apoptosis showed that compared with those in the sham operation group,the numbers of apoptotic cells and the apoptosis index were significantly increased in the model group;compared with those in the model group,the apoptotic cells in the spinal cord injury site and the apoptosis index were significantly decreased in the fire needling group(P<0.05,P<0.01),and the apoptosis index reached the lowest on 14 d in the fire needling group.Compared with that in the sham operation group,the relative protein expression of DAP12 tended to increase at each time point in the model group,but the difference was not statistically significant(P>0.05);compared with that in the model group,the relative protein expression of DAP12 tended to increolse in the fire needling group,and the increase was most obvious on 7 d,but the differences were not statistically significant(P>0.05).Compared with those in the sham operation group,the relative protein expression of P-PI3K tended to increase at all time points in the model group,and the difference was only statistically significant on 1 d(P<0.05);compared with those in the model group,the relative protein expresion of p-PI3K was decreased on 1 d,3 d and 14 d,whereas the relative protein expression of p-PI3K was significantly increased on 7 d in the fire needling group(P<0.05).Compared with that in the sham operation group,the protein expression of TGF-β was increased in the model group on 1 d,7 d and 14 d,and the difference was only statistically significant on 14 d(P<0.05);compared with that in the model group,the protein expression of TGF-β was higher in the fire needling group on 3 d and 7 d,but the difference was only statistically significant on 3 d(P<0.01).Conclusion:Fire needling can regulate the protein expressions of p-PI3K,TGF-β and DAP12 after SCI,thereby improving the apoptosis of neurocytes and promoting the recovery of nerve motor function after SCI.
作者
李璨
赵志恒
徐家淳
焦召华
王凯
LI Can;ZHAO Zhiheng△;XU Jiachun;JIAO Zhaohua;WANG Kai(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300250,China;Tianjin Gong'an Hospital,Tianjin 300040,China)
出处
《针灸临床杂志》
2023年第5期70-76,共7页
Journal of Clinical Acupuncture and Moxibustion
基金
国家自然科学基金青年项目,编号:81903997
天津市教委科研计划,编号:2022KJ181。
