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瞿启睿, 孙晓莹, 邓石峰, 许明, 祁芳, 易细芹, 唐丽亚, 卓越, 艾坤.电针对T10脊髓损伤后T10-L2脊髓组织蛋白质组学的生物信息学分析[J].湖南中医药大学学报英文版,2025,45(9):1670-1680.[Click to copy
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| 电针对T10脊髓损伤后T10-L2脊髓组织蛋白质组学的生物信息学分析 |
| 瞿启睿,孙晓莹,邓石峰,许明,祁芳,易细芹,唐丽亚,卓越,艾坤 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 运用串联质量标签（TMT）定量蛋白质组学生物信息学分析技术，探讨电针治疗T10脊髓损伤后神经源性膀胱的可能机制。方法 将60只雌性成年SD大鼠随机分为假手术组（12只）、造模组（48只）。造模组大鼠采用Hassan Shaker脊髓横断法制备T10节段骶上脊髓损伤模型，脊髓休克期过后共存活且符合成模标准共22只，二次随机分为模型组和电针组，每组11只。假手术组随机抽取11只纳入实验。电针组在造模后第19天于次髎、中极、三阴交给予电针干预，20 min/次，1次/d，连续干预10 d。干预结束后，大鼠行尿流动力学和HE染色检测，取T10-L2脊髓组织行TMT定量蛋白质组学检测和Western blot检测。结果 与假手术组相比，模型组大鼠漏尿点压力（LPP）和膀胱最大容量（MCC）明显增高（P<0.01）；与模型组相比，电针组大鼠LPP和MCC明显降低（P<0.01）。膀胱组织HE染色显示，模型组膀胱颈组织大量炎症细胞浸润，弹性纤维减少，肌层平滑肌纤维排列略紊乱；电针组膀胱颈组织炎症细胞浸润减少，平滑肌纤维排列趋于整齐。TMT定量蛋白质组学显示，T10-L2脊髓组织中，模型组与假手术组相比，筛选出146个差异表达蛋白（DEPs），电针组与模型组相比，筛选出37个DEPs，经电针治疗后共有5个DEPs被反向调节。经生物信息学分析，提示电针对损伤局部脊髓组织的影响可能涉及调节炎症反应、氧化应激、细胞凋亡、神经保护及轴突再生。脊髓组织HE染色显示，模型组T10-L2脊髓组织有神经元崩解现象，坏死后空洞增多，炎症细胞增生；电针组神经元崩解现象减少，水肿减轻，炎症细胞相对减少。与假手术组相比，模型组大鼠T10-L2脊髓组织富组氨酸糖蛋白（Hrg）、胎儿素B(Fetub)表达明显升高（P<0.01），而G蛋白亚基γ4(Gng4)表达明显降低（P<0.01）；与模型组相比，电针组大鼠T10-L2脊髓组织Hrg、Fetub表达明显降低（P<0.01），而Gng4表达明显升高（P<0.01）。结论 电针次髎、中极、三阴交可有效减轻T10脊髓损伤后的继发性损伤，保护神经组织，抑制炎症反应、氧化应激与细胞凋亡，激活神经保护及轴突再生相关通路，降低膀胱颈阻力并提高排尿效率。 |
| 关键词: 脊髓损伤 蛋白质组学 生物信息学 胎儿素B 富组氨酸糖蛋白 G蛋白亚基γ4 |
| DOI：10.3969/j.issn.1674-070X.2025.09.011 |
| Received:April 03, 2025 |
| 基金项目:国家自然科学基金项目(81874510); 湖南中医药大学研究生创新项目(2023CX74); 2023年湖南省教育厅科学研究优秀青年项目(23B0350)。 |
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| Bioinformatic analysis of proteomics in T10-L2 spinal cord tissue following electroacupuncture treatment for T10 spinal cord injury |
| QU Qirui, SUN Xiaoying, DENG Shifeng, XU Ming, QI Fang, YI Xiqin, TANG Liya, ZHUO Yue, AI Kun |
| (School of Acupuncture-moxibustion, Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To investigate the potential mechanism of electroacupuncture（EA） in treating neurogenic bladder（NB）after T10 spinal cord injury（SCI） using tandem mass tag（TMT）-based quantitative proteomics and bioinformatic analysis. Methods Sixty female adult SD rats were randomly divided into sham-operated group（n=12） and modeling group（n=48）. A suprasacral SCI model at the T10 segment was established in the modeling group using the Hassan Shaker spinal cord transection method. After the spinal shock period, 22 surviving rats that met the modeling criteria were further randomized into model group（n=11） and EA group（n=11）. Eleven rats from the sham-operated group were randomly selected for inclusion. The EA group was treated with EA at the acupoints such as Ciliao（BL32）, Zhongji（CV3）, and Sanyinjiao（SP6） starting on day 19 post-modeling. The intervention was performed once daily for 20 minutes per session, lasting for 10 consecutive days. After intervention, urodynamic testing and HE staining were performed. T10-L2 spinal cord tissue was collected for TMT-based quantitative proteomics and Western blot analysis. Results Compared with the sham-operated group, the model group exhibited significantly increased leak point pressure（LPP） and maximum cystometric capacity（MCC）（P＜0.01）. Compared with the model group, the EA group showed significantly reduced LPP and MCC（P＜0.01）. HE staining of bladder neck tissue revealed marked inflammatory cell infiltration, reduced elastic fibers, and slightly disordered arrangement of smooth muscle fibers in the model group, while the EA group showed decreased inflammatory cell infiltration and more orderly smooth muscle fiber arrangement. TMT-based quantitative proteomics of T10-L2 spinal cord tissue identified 146 differentially expressed proteins（DEPs） between the model and sham-operated groups, and 37 DEPs between the EA and model groups, among which five DEPs were reversely regulated after EA intervention. Bioinformatic analysis indicated that EA may modulate inflammatory response, oxidative stress, apoptosis, neuroprotection, and axonal regeneration in the injured local spinal cord. HE staining of T10-L2 spinal cord tissue showed neuronal disintegration, increased post-necrotic cavities, and inflammatory cell proliferation in the model group, while the EA group exhibited reduced neuronal disintegration, alleviated edema, and fewer inflammatory cells. Compared with the sham-operated group, the model group showed significantly upregulated expression of histidine-rich glycoprotein（Hrg） and fetuin-B（Fetub）（P＜0.01）, and downregulated expression of G protein subunit gamma 4（Gng4） in the T10-L2 spinal cord tissue（P＜0.01）. Compared with the model group, the EA group showed significantly downregulated expression of Hrg and Fetub in the T10-L2 spinal cord tissue（P＜0.01）, and significantly upregulated expression of Gng4（P＜0.01）. Conclusion EA at Ciliao（BL32）, Zhongji（CV3）, and Sanyinjiao（SP6） acupoints can effectively alleviate secondary injury after T10SCI, protect neural tissue, suppress inflammatory response, oxidative stress, and apoptosis, and activate neuroprotection and axonal regeneration pathways, thereby reducing bladder neck resistance and improving voiding efficiency. |
| Key words: spinal cord injury proteomics bioinformatics fetuin-B histidine-rich glycoprotein G protein subunit gamma 4 |
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