Man H, Wang J, Wu M, Shao Y, Yang J, Li S, Lü J, Zhou Y
Nanjing University of Chinese Medicine, Nanjing 210023, China.
Wuxi Traditional Chinese Medicine Hospital, Wuxi 214071, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Apr 20;44(4):636-643. doi: 10.12122/j.issn.1673-4254.2024.04.03.
To investigate the effect of formula-medicated serum for promoting spinal cord injury (SCI) repair in rats and explore the possible mechanism.
Thirty adult SD rats were randomized into sham-operated group, SCI (induced using a modified Allen method) model group, and formula-medicated serum treatment group. After the operations, the rats were treated with normal saline or by gavage on a daily basis for 14 days, and the changes in hindlimb motor function of the rats was assessed with Basso-Beattie-Bresnahan (BBB) scores and inclined-plate test. The injured spinal cord tissues were sampled from the SCI rat models for single-cell RNA sequencing, and bioinformatics analysis was performed to identify the target genes of , spinal cord injury and glycolysis. In the cell experiment, cultured astrocytes from neonatal SD rat cortex were treated with SOX2 alone or in combination with -medicated serum for 21 days, and the protein expressions of PKM2, p-PKM2 and YAP and colocalization of PKM2 and YAP in the cells were analyzed with Western blotting and immunofluorescence staining, respectively.
The SCI rats with treatment showed significantly improved BBB scores and performance in inclined-plate test. At the injury site, high PKM2 expression was detected in various cell types. Bioinformatic analysis identified the HIPPO-YAP signaling pathway as the target pathway of . In cultured astrocytes, SOX2 combined with the mediated serum, as compared with SOX2 alone, significantly increased PKM2, p-PKM2 and YAP expressions and entry of phosphorylated PKM2 into the nucleus, and promoted PKM2 and YAP co-localization in the cells.
formula accelerates SCI repair in rats possibly by promoting aerobic glycolysis of the astrocytes activating the PKM2/YAP axis to induce reprogramming of the astrocytes into neurons.
研究含药血清对大鼠脊髓损伤（SCI）修复的影响并探讨其可能机制。
将30只成年SD大鼠随机分为假手术组、SCI（采用改良Allen法诱导）模型组和含药血清治疗组。术后，大鼠每日经生理盐水灌胃或给予含药血清，共14天，采用Basso-Beattie-Bresnahan（BBB）评分和斜板试验评估大鼠后肢运动功能变化。从SCI大鼠模型中采集损伤脊髓组织进行单细胞RNA测序，并进行生物信息学分析以鉴定含药血清、脊髓损伤和糖酵解的靶基因。在细胞实验中，将新生SD大鼠皮质培养的星形胶质细胞单独用SOX2或与含药血清联合处理21天，分别用蛋白质免疫印迹法和免疫荧光染色法分析细胞中PKM2、p-PKM2和YAP的蛋白表达以及PKM2与YAP的共定位情况。
接受含药血清治疗的SCI大鼠BBB评分和斜板试验表现显著改善。在损伤部位，多种细胞类型中检测到高表达的PKM2。生物信息学分析确定HIPPO-YAP信号通路为含药血清的靶通路。在培养的星形胶质细胞中，与单独使用SOX2相比，SOX2联合含药血清显著增加PKM2、p-PKM2和YAP的表达以及磷酸化PKM2进入细胞核，并促进细胞中PKM2与YAP的共定位。
含药血清可能通过促进星形胶质细胞有氧糖酵解、激活PKM2/YAP轴诱导星形胶质细胞重编程为神经元，从而加速大鼠SCI修复。
