| 基于TNFR/RIPK1通路探索紫草素对大鼠脊髓损伤的作用及机制 |
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Received:October 19, 2023
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| 作者 | Author | 单位 | Unit | E-Mail |
| 史吉胜 |
SHI Ji-sheng |
山东大学齐鲁医院德州医院骨科, 山东 德州 253600 |
Department of Orthopedics, Qilu Hospital of Shandong University Dezhou Hospital, Dezhou 263000, Shandong, China |
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| 钱吉泽 |
QINA Ji-ze |
山东大学齐鲁医院德州医院骨科, 山东 德州 253600 |
Department of Orthopedics, Qilu Hospital of Shandong University Dezhou Hospital, Dezhou 263000, Shandong, China |
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| 王金广 |
WANG Jin-guang |
山东大学齐鲁医院德州医院骨科, 山东 德州 253600 |
Department of Orthopedics, Qilu Hospital of Shandong University Dezhou Hospital, Dezhou 263000, Shandong, China |
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| 林斌 |
LIN Bin |
中国人民解放军第909医院骨科, 福建 漳州 363000 |
Department of Orthopedics, the 909 th Hospital of PLA, Zhangzhou 363000, Fujian, China |
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| 庞同涛 |
PANG Tong-tao |
山东大学齐鲁医院德州医院骨科, 山东 德州 253600 |
Department of Orthopedics, Qilu Hospital of Shandong University Dezhou Hospital, Dezhou 263000, Shandong, China |
754681652@qq.com |
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| 期刊信息：《中国骨伤》2024年37卷，第1期，第61-68页 |
| DOI：10.12200/j.issn.1003-0034.20230567 |
| 基金项目:山东省中医药科技项目(编号:2020Q119) |
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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、21 d行斜板实验,再处死动物取脊髓组织。造模后1 h 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)蛋白及凋亡相关蛋白受体相互作用蛋白激酶1(receptor-interacting protein kinase 1,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3 d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24 h,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。 |
| [关键词]:脊髓损伤 紫草素 功能恢复 |
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| Mechanism of Shikonin on spinal cord injury in rats based on TNFR/RIPK1 pathway |
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Abstract:
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. |
| KEYWORDS:Spinal cord injury Shikonin Nerve repair |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 史吉胜,钱吉泽,王金广,林斌,庞同涛.基于TNFR/RIPK1通路探索紫草素对大鼠脊髓损伤的作用及机制[J].中国骨伤,2024,37(1):61~68 |
| 英文格式: | SHI Ji-sheng,QINA Ji-ze,WANG Jin-guang,LIN Bin,PANG Tong-tao.Mechanism of Shikonin on spinal cord injury in rats based on TNFR/RIPK1 pathway[J].zhongguo gu shang / China J Orthop Trauma ,2024,37(1):61~68 |
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