摘要
目的:初步探讨紫草素对大鼠急性脊髓损伤(spinal cord injury,SCI)后神经功能恢复的影响及作用机制。方法:将96只Sprague-Dawley(SD)雄性大鼠分为4组:假手术组,即A组;假手术+紫草素组,即B组;脊髓损伤+二甲基亚砜(dimethyl sulfoxide,DMSO)组,即C组;脊髓损伤+紫草素组,即D组;每组24只。C、D组采用钳夹法制作大鼠急性SCI模型。所有大鼠硬膜下置管,A组不给药,B组和D组造模后30 min经导管注射紫草素100 mg·kg^(-1),C组注射等量DMSO,每日1次,至取材时间点。各组分别于造模后6、12 h和3 d每组取8只大鼠,行Basso-Beattie-Bresnahan(BBB)评分及造模后1、3、7、14、21d行斜板实验,再处死动物取脊髓组织。造模后1h每组大鼠腹腔注射碘化丙啶(propidine iodide,PI)1 mg·kg^(-1),术后24 h取材检测脊髓组织PI红染细胞数;24 h时取材采用苏木素-伊红(haematoxylin eosin,HE)染色观察脊髓损伤情况,尼氏(Nissl)染色观察神经元存活数量,使用Western-blot技术检测B细胞淋巴瘤-2(B cell lymphoma-2,Bcl-2)蛋白及调亡相关蛋白受体相互作用蛋白激酶1(receptor-interacting protein kinase1,RIPK1)的表达水平。结果:造模后A组和B组各时间点的BBB评分均正常,C、D组各时间点均低于A、B组,D组造模后12 h和3d的BBB评分高于同时间点C组(P<0.05)。造模后12 h,D组PI红染细胞较C组明显减少,神经元崩解减轻(P<0.05)。造模后24h,A组和B组脊髓组织HE和Nissl染色正常,D组脊髓组织损伤程度和存活神经元数量均优于C组(P<0.05)。Bcl-2、RIPK1蛋白在A组、B组表达很低;RIPK1在C组表达明显增高,在D组表达明显下降,差异有统计学意义(P<0.05);Bcl-2蛋白在D组表达高于C组(P<0.05)。结论:紫草素可减轻大鼠急性SCI后的病理变化,改善行为学评分,促进脊髓神经功能恢复。其具体机制可能与抑制TNFR/RIPK1信号通路介导的坏死性凋亡有关。
Objective To explore the effect of shikonin on the recovery of nerve function after acute spinal cord injury(SCI)in rats.Methods 96 male Sprague-Dawley(SD)rats were divided into 4 groups randomly:sham operation group(Group A),sham operation+shikonin group(Group B),SCI+DMSO(Group C),SCI+shikonin group(Group D).The acute SCI model of rats was made by clamp method in groups C and D.After subdural catheterization,no drug was given in group A.rats in groups B and D were injected with 100 mg·kg^(-1)of shikonin through catheter 30 min after modeling,and rats in group C were given with the same amount of DMSO,once a day until the time point of collection tissue.Basso-Beattie-Bresnahan(BBB)scores were performed on 8 rats in each group at 6,12,and 3 d after moneling,and oblique plate tests were performed on 1,3,7 and 14 d after modeling,and then spinal cord tissues were collected.Eight rats were intraperitoneally injected with propidine iodide(PI)1 h before sacrificed to detection PI positive cells at 24 h in each group.Eight rats were sacrificed in each group at 24 h after modeling,the spinal cord injury was observed by HE staining.The Nissl staining was used to observe survivor number of nerve cells.Western-blot technique was used to detect the expression levels of Bcl-2 protein and apoptosis related protein RIPK1.Results After modeling,BBB scores were normal in group A and B,but in group C and D were significantly higher than those in group A and B.And the scores in group D were higher than those in group C in each time point(P<0.05).At 12 h after modeling,the PI red stained cells in group D were significantly reduced compared with that in group C,and the disintegration of neurons was alleviated(P<0.05).HE and Nissl staining showed nerve cells with normal morphology in group A and B at 24h after operation.The degree of SCI and the number of neuronal survival in group D were better than those in group C,the difference was statistically significant at 24h(P<0.05).The expression of Bcl-2 and RIPK1 proteins was very low in group A and B;The expression of RIPK1 was significantly increased in Group C and decreased in Group D,with a statistically significant difference(P<0.05);The expression of Bcl-2 protein in group D was significantly higher than that in group C(P<0.05).Conclusion Shikonin can alleviate the pathological changes after acute SCI in rats,improve the behavioral score,and promote the recovery of spinal nerve function.The specific mechanism may be related to the inhibition of TNFR/RIPK1 signaling pathway mediated necrotic apoptosis.
作者
史吉胜
钱吉泽
王金广
林斌
庞同涛
SHI Ji-sheng;QINA Ji-ze;WANG Jin-guang;LIN Bin;PANG Tong-tao(Department of Orthopedics,Qilu Hospital of Shandong University Dezhou Hospital,Dezhou 263000,Shandong,China;Department of Orthopedics,the 909th Hospital of PLA,Zhangzhou 363000,Fujian,China)
出处
《中国骨伤》
CAS
CSCD
2024年第1期61-68,共8页
China Journal of Orthopaedics and Traumatology
基金
山东省中医药科技项目(编号:2020Q119)。
关键词
脊髓损伤
紫草素
功能恢复
Spinal cord injury
Shikonin
Nerve repair
