脊髓康对大鼠脊髓损伤区小胶质细胞/神经元的影响

Effect of JiSuiKang on the microglia infiltration and neuron survival in the spinal cord injury area of rats

目的:观察中药复方脊髓康(JSK)对大鼠脊髓损伤区域小胶质细胞浸润与神经元存活情况的影响。方法:60只SPF级SD大鼠,随机分为假手术组(Sham)、模型组(Ctrl)、JSK低、中、高剂量组(JSK-L、JSK-M、JSK-H)、强的松组;改良Allen's法建立大鼠脊髓损伤(spinal cord injury,SCI)模型;术后各组以不同的方式干预,行大鼠双后肢运动功能评分(BBB评分);酶联免疫吸附法(ELISA)检测血清炎症因子IL-1表达,损伤区脊髓组织行冰冻切片,CD11b/c、β3-Tubulin免疫荧光染色,观察小胶质细胞的浸润情况与神经元的生长情况。结果:在BBB评分方面:Sham、JSK-L、JSK-M、JSK-H、强的松组均高于Ctrl(P<0.05);在ELISA检测方面,Sham、JSK-L、JSK-M、JSK-H、强的松组的IL-1表达均低于Ctrl(P<0.05),强的松组、JSK-H低于JSK-L、JSK-M(P<0.05);在小胶质细胞浸润百分比方面,JSK-L、JSK-M、JSK-H、强的松组均低于Ctrl(P<0.05),JSK-H与强的松组相比差异无统计学意义(P>0.05)。从神经元的生长状态上:JSK-L、JSK-M、JSK-H、强的松组优于Ctrl(P<0.01);JSK-H优于JSK-L、JSK-M(P<0.05)。结论:中药复方JSK抑制SCI后体内小胶质细胞的迁移浸润,促进受损神经元的恢复,进而促进SCI大鼠双下肢运功功能的恢复。

AIM:To observe the effect of Chinese medicine compound JiSuiKang on the microglia infiltration and neuron survival at the spinal cord injury area of rats.METHODS:Sixty SD rats were randomly divided into sham operation group(Sham),model group(Ctrl),JSK low(JSK-L),JSK medium(JSK-M)and JSK high(JSK-H)dose group,prednisone group.The model of rat spinal cord injury(SCI)was established by improved Allen's method.After the operation,each group was intervened in different ways,and then we assessed the double hind limb motor function score in rats(BBB score).The heart perfusion was done and the spinal cord tissues from the injury area were taken.ELISA tests were used to detect the expression of inflammatory factors.The spinal cord tissue was sliced into sections and then immunofluorescence stained of CD11b/c and beta 3-Tubulin to observe the infiltration of microglia and the growth of neurons under the inverted fluorescence microscope.RESULTS:The BBB score of Sham,JSK-L,JSK-M,JSK-H and prednisone group were higher than Ctrl(P<0.05).For the expression of inflammation,the expressions of IL-1 in group Sham,JSK-L,JSK-M,JSK-H and prednisone were lower than that of Ctrl(P<0.05).For the percentage of microglia infiltration,JSK-L,JSK-M,JSK-H and prednisone were all lower than that of Ctrl(P<0.05),and there was no significant difference between JSK-H and prednisone(P>0.05).For the growth state of the neuron,the JSK-L,JSK-M,JSK-H,prednisone group were better than Ctrl(P<0.01),and JSK-H was better than JSK-L and JSK-M(P<0.05).CONCLUSION:The Chinese medicine compound JSK inhibits the migration and invasion of microglia in SCI,promotes the recovery of damaged neurons and motor function in SCI rats.