大鼠压疮局部不同温度治疗对PI3K/Akt/GSK3β 信号通路的影响

Effect of different local temperature on PI3K/Akt/GSK3β pathway in rat model of pressure ulcer

摘要:

目的 探讨不同局部温度对大鼠压疮PI3K/Akt/GSK3β 通路的影响，为压疮的临床治疗提供基础实验依据。方法 用缺血再灌注（IRI）法复制大鼠压疮模型。将40 例无特定病原体级（SPF）健康成年雄性SD 大鼠随机分为4 组：假手术组（Sham 组）、常温IRI 组（NTI）、低温IRI 组（LTI）和高温IRI 组（HTI）。HE 染色观察骨骼肌病理变化；Western blot 检测Akt、GSK3β 的磷酸化情况；免疫荧光观察p-Akt、p-GSK3β 的表达情况。TUNEL 检测大鼠压疮组织细胞凋亡情况。结果 与常温IRI 组比较，高温IRI 组的骨骼肌细胞病理损伤加重，凋亡细胞增加，Western blot 和免疫荧光结果显示Akt 和GSK3β 的磷酸化水平均降低（P <0.05）；而低温IRI 组骨骼肌细胞病理损伤减轻，Western blot 和免疫荧光结果显示Akt 和GSK3β 的磷酸化水平均上升（P <0.05）。结论 局部高温能抑制Akt 和GSK3β 的磷酸化水平而加重大鼠压疮损伤，局部低温能提高Akt 和GSK3β 的磷酸化水平减轻大鼠压疮损伤。

Abstract:

Objectives To investigate the effect of different local temperature on PI3K/Akt/GSK3β pathway in rat model of pressure ulcer. Methods The rat model of pressure ulcer was established by ischemia reperfusion(IR) method. 40 SD rats of SPF grade were divided into Sham group, NTI group, HTI group and LTI group.HE staining was performed to observe pathologies of skeletal muscle. Western blot analysis was utilized to determine the phosphorylation of Akt and GSK3β. Expression levels of p-Akt and p-GSK3β were determined by Immunofluorescence; TUNEL staining was performed to identify apoptosis rate of skeletal muscle. Results Morphological abnormalities of skeletal muscle, apoptosis rate increased while phosphorylation level of Akt and GSK3β decreased in HTI group when compared with NTI group (P < 0.05). In contrast, low temperature can restrain the skeletal muscle cell damage and apoptosis, and increased the phosphorylation of Akt and GSK3β when compared with NTI group (P < 0.05). Conclusions Local hypothermia can enhance recovery of pressure ulcer injury by increasing the phosphorylation of Akt and GSK3β, and this can provide experimental evidence for clinical usage of hypothermia for treatment of pressure ulcer.