小分子融合肽TAT-KIR调节创伤性脑损伤后星形胶质细胞激活加速神经功能修复

ZHANG  Hou-ying1; CHENG  Li-xue1; FENG  Yun-ran1; 2; WANG  Xin-yi1; WU  Han-xi1; CAO  Wei-zhong1; YANG  Shu-han1; SUN  Hao-ran1; SHU  Zhang1; MA  Kai-ge1*; HU  Xiao-xuan1; 3

doi:10.16098/j.issn.0529-1356.2026.04.001

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解剖学报 ›› 2026, Vol. 57 ›› Issue (4) : 391-399. DOI: 10.16098/j.issn.0529-1356.2026.04.001

神经生物学

小分子融合肽TAT-KIR调节创伤性脑损伤后星形胶质细胞激活加速神经功能修复

张厚莹¹程立雪¹冯韵然^1,2王心怡¹吴晗溪¹曹为众¹杨舒涵¹孙浩然¹舒章¹马凯歌^1*胡晓宣^{1,3 *}

作者信息 +

Small-molecule fusion peptide TAT-KIR improving neurofunctional recovery by regulating astrocytes activation in mice after traumatic brain injury

ZHANG Hou-ying¹, CHENG Li-xue¹, FENG Yun-ran^1,2, WANG Xin-yi¹, WU Han-xi¹, CAO Wei-zhong¹, YANG Shu-han¹, SUN Hao-ran¹, SHU Zhang¹, MA Kai-ge^1*, HU Xiao-xuan^1,3 *

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摘要

目的探讨创伤性脑损伤（TBI）后小分子融合肽转录反式激活因子（TAT）-细胞因子信号抑制物3（SOCS3）蛋白的激酶抑制区（KIR）调节星形胶质细胞激活，促进损伤修复的作用。方法将2月龄雄性C57BL/6小鼠随机分为假手术组(sham)、损伤组(TBI)和治疗组(TAT-KIR)，每组15只。TBI组使用Feeney自由落体法构建TBI模型，TAT-KIR组在TBI后立即原位注射TAT-KIR，sham组只开颅不打击。TBI后1 d Western blotting检测小鼠皮质损伤区及周围组织JAK2/STAT3通路激活情况；TBI后1、3、7及14 d，NSS和Beam walking评估小鼠运动功能，免疫荧光染色检测星形胶质细胞和神经元数量，并使用Sholl analysis对星形胶质细胞形态进行分析；TBI后3 d和7 d，Real-time PCR检测星形胶质细胞A1型标志物C3、H2-D1、Serping1和H2-T23和A2型标志物转谷氨酰胺酶1(Tgm-1)、心肌营养素样细胞因子 1（Clcf1）、S100 钙结合蛋白 A10 (S100a10) 和CD109基因表达及相关因子水平，ELISA检测局部脑组织谷氨酸含量。结果 TAT-KIR治疗组小鼠激活的JAK2/STAT3信号通路被抑制，皮质损伤区反应性星形胶质细胞增生减弱且分支变少；C3、H2-D1、Serping1、H2-T23（A1型）表达下调，Tgm-1、Clcf1、CD109、S100a10（A2型）表达上调；促突触生成因子（Gpc4、Sparcl1、Thbs1、Thbs2）和神经营养因子［脑源性神经营养因子（BDNF）、睫状神经营养因子（CNTF）、胶质细胞源性神经营养因子（GDNF）、神经营养素-3 （NT3）、神经生长因子（NGF）］ mRNA表达水平升高，谷氨酸转运蛋白1（GLT-1）和谷氨酸/天冬氨酸转运蛋白（GLAST）表达增多，皮质损伤区组织中谷氨酸含量降低；随恢复时间增加，TBI小鼠皮质损伤区神经元丢失明显减轻，NSS评分降低，平衡木移动距离增加。结论 TBI急性期给予TAT-KIR，可有效减弱星形胶质细胞反应性增生，增加A2型细胞比例和神经保护功能，加速小鼠神经功能恢复。

Abstract

ObjectiveTo investigate the therapeutic potential of the small-molecule fusion peptide trans-activator of transcription（TAT）-kinase inhibitory region（KIR）of cytokine signaling 3（SOCS3） protein in mice by modulating astrocyte activation after traumatic brain injury (TBI). Methods Male C57BL/6 mice (2-month-old) were randomly divided into three groups, sham group (sham), TBI group (TBI), and TAT-KIR group (TAT-KIR), and each group had 15 mice. The TBI group received traumatic brain injury using Feeney’s free-fall method, the TAT-KIR group was injected with TAT-KIR in situ right after TBI, the sham group underwent craniotomy without impact. Western blotting was used to assess the activation of JAK2/STAT3 pathway at day 1 post-TBI. Immunofluorescence staining was employed to evaluate the astrocyte activation and the neuron loss at day 1, 3, 7, and 14 post-TBI, and Sholl analysis was used to analyze the morphology of astrocytes. Meanwhile, neurological severity score (NSS) and beam walking tests were used to evaluate motor function. At day 3 and 7 post-TBI, Real-time PCR was performed to measure the mRNA expression levels of C3, H2-D1, Serping1, H2-T23 (A1-astrocyte) and transglutaminase 1(Tgm-1), cardiotrophin-like cytokine factor 1（Clcf1）, S100 calcium-binding protein A10(S100a10), CD109 (A2-astrocyte), and the cytokines secreted by activated astrocytes. ELISA was used to detecte the amount of glutamate in injured cortex. Results In the TAT-KIR treated group, TBI-induced activation of the JAK2/STAT3 signaling pathway and astrogliosis were suppressed. The branches of activated astrocytes decreased. The mRNA level of C3, H2-D1, Serping1, H2-T23 were downregulated, while the mRNA level of Tgm-1, Clcf1, CD109, S100a10 were upregulated. At the same time, the synaptogenic factors, including Gpc4, Sparcl1, Thbs1, Thbs2, and the neurotrophic factors, such as brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), glial cell line-derived neurotrophic factor (GDNF), neurotrophin-3 (NT3), nerve growth gactor (NGF) were significantly upregulated. The expression of glutamate transporters, including glutamate transporter 1（GLT-1）and glutamate/aspartate transporter（GLAST）increased, while the amount of extracellular glutamate decreased. The neuron loss of TBI mice gradually reduced with the time. NSS scores decreased and the distance of beam walking significantly increased. Conclusion Administration of TAT-KIR during the acute phase of TBI reduced astrocyte activation, upregulated the proportion of A2 astrocyte, and subsequently accelerated neurological function recovery in TBI mice.

导出引用

张厚莹程立雪冯韵然王心怡吴晗溪曹为众杨舒涵孙浩然舒章马凯歌胡晓宣. 小分子融合肽TAT-KIR调节创伤性脑损伤后星形胶质细胞激活加速神经功能修复[J]. 解剖学报. 2026, 57(4): 391-399 https://doi.org/10.16098/j.issn.0529-1356.2026.04.001

ZHANG Hou-ying, CHENG Li-xue, FENG Yun-ran, WANG Xin-yi, WU Han-xi, CAO Wei-zhong, YANG Shu-han, SUN Hao-ran, SHU Zhang, MA Kai-ge, HU Xiao-xuan. Small-molecule fusion peptide TAT-KIR improving neurofunctional recovery by regulating astrocytes activation in mice after traumatic brain injury[J]. Acta Anatomica Sinica. 2026, 57(4): 391-399 https://doi.org/10.16098/j.issn.0529-1356.2026.04.001

中图分类号： R322.81

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