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- 2018
TGN-020对大鼠脊髓损伤后继发性水肿和星形胶质细胞增生的影响
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Abstract:
摘要：目的 检测水通道蛋白4(aquaporin 4, AQP4)特异性抑制剂TGN-020对大鼠脊髓损伤(spinal cord injury, SCI)后继发性水肿和星形胶质细胞增生相关指标的影响，为SCI后水肿与星形细胞增生之间的关系研究提供重要依据。方法 成年雌性SD大鼠72只，体质量180～220g，随机分为假手术组(Sham组) 、单纯脊髓损伤组(SCI组)、TGN-020干预组(TGN-020组)，每组24只。采用改良后的动脉夹挤压脊髓建立SCI模型，Sham组仅行椎板切除术。术后TGN-020组腹腔内注射TGN-020(5mg/kg，ip)处理，Sham组和SCI组予以等体积的二甲基亚砜(dimethyl sulfoxide, DMSO)各组于术后3d收集标本，分别采用干湿重法检测含水量变化，采用蛋白印记、免疫荧光检测AQP4、胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)、增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)蛋白表达。结果 术后3d，与Sham组相比，SCI组和TGN-020组脊髓损伤区含水量明显升高(P<0.01)，与SCI组相比，TGN-020组脊髓损伤区含水量减少(P<0.05)；SCI组和TGN-020组AQP4、GFAP、PCNA的表达较Sham组明显升高(P<0.01)，TGN-020组AQP4、GFAP、PCNA的表达较SCI组降低(P<0.05)。结论 TGN-020可以通过下调大鼠脊髓损伤后AQP4的表达减轻继发性水肿，抑制星形胶质细胞增生，从而促进大鼠急性损伤后的功能恢复。
ABSTRACT: Objective To explore the effects of TGN-020 as a specific inhibitor of aquaporin 4 (AQP4) on secondary edema and astrocyte proliferation after spinal cord injury (SCI) in rats so as to provide important basis for studies on the relationship between edema and astrocyte proliferation after SCI. Methods Seventy-two adult female SD rats (180-220g) were randomly chosen and divided into three groups (n=24): sham-operation group (sham group), spinal cord injury group (SCI group), and TGN-020 intervention group (TGN-020 group). We used the modified artery clip and squeezed spinal cord to establish the model of SCI, sham group underwent lamnectomy without the spinal cord injuried. TGN-020 group received TGN-020 (5mg/kg, ip); dimethyl sulfoxide (DMSO) of the same volume with TGN-020 group was injected into SCI group and sham group via intraperitoneal injection postoperatively. Then water content and the expressions of AQP4, glial fibrillary acidic protein (GFAP) and proliferating cell nuclear antigen (PCNA) were detected with wet-dry weighting method and Western blot, immunofluorescence staining in three days respectively after SCI. Results Three days after operation, the water content of spinal cord injury area in SCI group and TGN-020 group was significantly higher than that in sham group (P<0.01). The water content of spinal cord injury area in TGN-020 group was significantly lower than that in SCI group (P<0.05). The expressions of AQP4, GFAP, and PCNA in SCI group and TGN-020 group were significantly higher than those in sham group (P<0.01). The expressions of AQP4, GFAP, and PCNA were significantly lower in TGN-020 group than in SCI group (P<0.05). Conclusion TGN-020 can reduce secondary edema and inhibit astrocyte proliferation by down-regulating the expression of AQP4 after SCI in rats, which can promote
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