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大鼠早期压疮缺血再灌注损伤的凋亡机制及曲美他嗪预处理的效果

发布时间:2018-10-08 14:17  文章来源:笔耕文化传播
【摘要】:目的探讨大鼠早期压疮缺血再灌注损伤的凋亡机制及曲美他嗪预处理的效果,为临床早期压疮的防治提供基础理论依据和新思路。方法1将16只Wistar大鼠随机分为2组,对照组(Sham组)和缺血/再灌注组(I/R组)。I/R组大鼠应用特制压力装置制备大鼠早期压疮缺血再灌注损伤模型。实验终点取受压局部肌肉组织,采用HE染色法观察大鼠受压肌肉组织的病理学变化;TUNEL法检测大鼠受压组织中肌细胞凋亡情况;免疫组织化学法检测受压肌肉组织中凋亡相关蛋白Bax与Bcl-2的表达情况;Western blot法检测凋亡相关蛋白Bax、Bcl-2和CHOP在受压组织中的表达水平。2将40只Wistar大鼠随机分为5组,对照组(Sham组)、缺血/再灌注组(I/R组)、曲美他嗪低剂量组(TMZ低组)、曲美他嗪中剂量组(TMZ中组)、曲美他嗪高剂量组(TMZ高组)。应用特制压力装置制备大鼠压疮缺血再灌注损伤模型。I/R组大鼠只造模不给药,曲美他嗪预处理组大鼠造模前7天分别给予曲美他嗪低(5mg/kg)、中(10mg/kg)、高(15mg/kg)剂量灌胃预处理。实验终点取受压局部肌肉组织,采用HE染色法观察各组大鼠受压肌肉组织的病理学变化;TUNEL法检测各组大鼠受压组织中肌细胞凋亡情况;免疫组织化学法检测各组受压肌肉组织中凋亡相关蛋白Bax与Bcl-2的表达情况;Western blot法检测凋亡相关蛋白Bax、Bcl-2和CHOP在受压组织中的表达水平。结果1光镜观察受压组织发现,Sham组肌纤维排列紧密,形态正常,无炎性细胞浸润及病理改变,与Sham组相比I/R组肌纤维病理损伤严重;TUNEL法检测受压组织凋亡结果显示,Sham组中几乎很少见到凋亡细胞,I/R组凋亡细胞的表达显著增多(P0.05);免疫组化检测Bax和Bcl-2蛋白结果与Western blot结果趋势一致,即Bax蛋白在Sham组中几乎不表达或有极少表达,与Sham组相比I/R组Bax蛋白表达显著升高(P0.05),Bcl-2蛋白在Sham组中表达显著,在I/R组中的表达与Sham组相比显著降低(P0.05);Western blot法检测受压组织中CHOP的表达结果显示,在Sham组中几乎不表达或仅有极少表达,与Sham组相比I/R组CHOP蛋白表达显著升高(P0.05)。2光镜观察受压组织发现,Sham组肌纤维排列紧密,形态正常,与Sham组相比I/R组肌纤维病理损伤严重,在曲美他嗪预处理组中随着给药剂量的增加病理损伤逐渐减轻。TUNEL法检测受压组织凋亡结果显示,Sham组中几乎很少见到凋亡细胞,I/R组凋亡细胞的表达显著增多(P0.05),在曲美他嗪预处理组中随着给药剂量的增加凋亡细胞表达逐渐减少。免疫组化检测Bax和Bcl-2蛋白结果与Western blot结果趋势一致,即Bax蛋白在Sham组中几乎不表达或有极少表达,与Sham组相比I/R组Bax蛋白表达显著升高(P0.05),在曲美他嗪预处理组中随着给药剂量的增加表达逐渐减少,Bcl-2蛋白在Sham组中表达显著,在I/R组中的表达与Sham组相比显著降低(P0.05),在曲美他嗪预处理组中随着给药剂量的增加表达逐渐增多。Western blot法检测受压组织中CHOP的表达结果显示,在Sham组中几乎不表达或仅有极少表达,与Sham组相比I/R组CHOP蛋白表达显著升高(P0.05),在曲美他嗪预处理组中随着给药剂量的增加表达逐渐减少。结论1细胞凋亡参与了大鼠早期压疮的形成,受压组织中Bcl-2蛋白表达减少,Bax和CHOP蛋白表达增多。2曲美他嗪预处理对大鼠早期压疮缺血再灌注损伤具有保护作用,可以减轻受压肌肉组织的病理损伤程度,上调Bcl-2表达水平,下调Bax、CHOP表达水平抑制细胞凋亡,保护受压肌肉组织。曲美他嗪高剂量(15mg/kg)对大鼠早期压疮缺血再灌注损伤的保护效果较低剂量好。
[Abstract]:Objective To investigate the mechanism of apoptosis in early stage pressure sore of rats and the pre-treatment effect of Qu Mei, and to provide theoretical basis and new thought for the prevention and cure of early stage pressure sore. Methods 16 Wistar rats were randomly divided into 2 groups, control group (sham group) and ischemia/ reperfusion group (I/ R group). A model of ischemia-reperfusion injury in rats with early pressure sore was prepared by using special pressure device in group I/ R rats. The results showed that the expression of Bax and Bcl-2 in compressed muscle tissue was detected by HE staining, and the expression of Bax and Bcl-2 was detected by immunohistochemical method. The expression levels of Bax, Bcl-2 and CHOP in compressed tissues were detected by Western blot. Forty Wistar rats were randomly divided into 5 groups, the control group (sham group), ischemia/ reperfusion group (I/ R group), the low-dose group (TMZ low group), and the dose group (group TMZ in TMZ). He was a high dose group (TMZ high). A model of ischemia-reperfusion injury in rats with pressure sore was prepared by using special pressure device. Rats in the I/ R group were not dosed, and the pre-treatment group was given a dose of low (5mg/ kg), medium (10mg/ kg) and high (15mg/ kg), respectively. At the end of the experiment, the local muscle tissue was compressed, and the pathological changes of muscle tissue were observed by HE staining, and the apoptosis was detected by TUNEL method. The expression of Bax, Bcl-2 and Bax, Bcl-2 and CHOP were detected by Western blot. Results According to light microscope, it was found that the muscle fibers of Shaam group were closely packed, the morphology was normal, there was no inflammatory cell infiltration and pathological change. Compared with the sham group, the pathological changes of myofibers in the I/ R group were serious. The expression of Bax and Bcl-2 protein in I/ R group was significantly increased (P0.05). The expression of Bcl-2 protein in group I/ R was significantly lower than that of sham group (P0.05). The expression of CHOP in compressed tissues was detected by Western blot. The expression of CHOP protein in group I/ R group was significantly higher than that of sham group (P0.05). The expression of apoptotic cells in the group I/ R was significantly increased (P0.05). The expression of Bax and Bcl-2 protein was similar to that of Western blot, that is, the expression of Bax protein was hardly expressed or expressed in the sham group, and the expression of Bax protein in the group I/ R was significantly higher than that of the sham group (P0.05). The expression of Bcl-2 protein in the sham group was significant, and the expression of Bcl-2 in the I/ R group was significantly lower than that of the sham group (P0.05). The expression of CHOP in compressed tissues was detected by Western blot. The expression of CHOP protein in group I/ R was significantly higher than that of sham group (P0.05). Conclusion 1 Apoptosis is involved in the formation of early pressure sore in rats, the expression of Bcl-2 protein in compressed tissues decreases, the expression of Bax and CHOP protein is increased. The expression level of Bcl-2 was up-regulated, the expression level of Bax and CHOP was down-regulated to inhibit cell apoptosis and to protect the compressed muscle tissue. The effect of high dose (15mg/ kg) on ischemia-reperfusion injury of early stage pressure sore in rats was lower than that of low dose.
【学位授予单位】:华北理工大学
【学位级别】:硕士
【学位授予年份】:2017
【分类号】:R47

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