摘要
目的比较大鼠压疮局部不同干预温度对内质网应激(ERS)及细胞凋亡的影响,为临床防治压疮提供实验依据。方法使用缺血再灌注法建立压疮模型,将40只SPF级成年雄性SD大鼠随机分成假手术组(sham组)、模型组、热干预组和冷干预组4组。在实验终点于冰上剪取各组大鼠受压部位肌肉组织,用HE染色观察骨骼肌病理学变化;Western blot检测内质网应激相关蛋白GRP78、caspase-12与CHOP的表达水平;免疫荧光观察caspase-12与CHOP的表达情况;TUNEL染色观察细胞凋亡情况。结果与模型组相比,热干预组的骨骼肌细胞损伤程度加重,凋亡细胞增多,免疫荧光结果显示caspase-12与CHOP的阳性表达增加,Western blot结果显示GRP78、caspase-12与CHOP的表达较模型组均上升(P<0.05);而冷干预组骨骼肌细胞损伤程度减轻,凋亡细胞减少,免疫荧光结果显示caspase-12与CHOP的阳性表达减少,Western blot结果显示GRP78、caspase-12与CHOP的表达较模型组均下降(P<0.05)。结论局部冷干预通过抑制内质网应激介导的凋亡通路而减轻大鼠压疮损伤;局部热干预加剧ERS而促进细胞凋亡的发生,加重大鼠压疮损伤。
Objective To compare the effects of different local intervention temperatures of pressure ulcer on endoplasmic reticulum stress（ ERS） and apoptosis in rats,and to provide an experimental evidence for clinical prevention and treatment of pressure ulcer. Methods The rat model of pressure ulcer was established by ischemia reperfusion,and a total of 40 SPF adult,male SD rats were divided into 4 groups： the sham group（ anesthesia only,without other treatment）,model group（ ischemia at 22℃ for 1 h and reperfusion at 22℃ as one cycle,repeated for 5 cycles）,high-temperature intervention group（ ischemia at 22℃ for 1 h and reperfusion at 32℃ as one cycle,repeated for 5 cycles） and lowtemperature intervention group（ ischemia at 22℃ for 1 h and reperfusion at 12℃ as one cycle,repeated for 5 cycles）. At the end of the experiment,muscle tissues at the sites under pressure of the rats were taken on ice. The pathological changes of skeletal muscle tissues were observed by HE staining. The expression levels of ERS-related proteins GRP78,caspase-12and CHOP were detected by Western blot, and the expression of caspase-12 and CHOP was also observed by immunofluorescence. Moreover,apoptosis in the skeletal muscle cells was examined by TUNEL staining. Results Compared with the model group,skeletal muscle cell damages became more severe and apoptotic cells were increased in the high-temperature intervention group. Besides,the results of the immunofluorescence assay showed an increased positive expression of caspase-12 and CHOP,and the results of Western blot showed that the expression levels of GRP78,caspase-12 and CHOP were all higher than those of the model group（ P〈0. 05）. In contrast,skeletal muscle cell damages were alliviated and apoptotic cells were reduced in the low-temperature intervention group. Meanwhile,the positive expression of caspase-12 and CHOP was decreased,as shown by immunofluorescence,and all the expression levels of GRP78,caspase-12 and CHOP detected by Western blot were lower than the control group（ P〈0. 05）. Conclusions Local lowtemperature intervention can alleviate the pressure ulcer damages in rats through inhibition of the ERS-mediated apoptotic pathway. Local high-temperature intervention may exacerbate the pressure ulcer damages in rats by activating the ERSmediated apoptotic pathway and promoting cell apoptosis. Local low-temperature intervention may be promising in clinical prevention and treatment of pressure ulcer.
出处
《中国比较医学杂志》
CAS
北大核心
2018年第1期56-63,共8页
Chinese Journal of Comparative Medicine
基金
华北理工大学研究生创新项目(编号:2017S30)
关键词
压疮
温度
缺血再灌注损伤
内质网应激
凋亡
大鼠
pressure ulcer
temperature
ischemia-reperfusion injury, IRI
endoplasmic reticulum stress, ERS
apoptosis
rats
