线粒体与内质网应激在压疮中的作用机制
发布时间：2018-12-08 12:14
【摘要】：目的探讨线粒体与内质网应激在大鼠压疮发生中的作用机制,线粒体及内质网应激的作用及其介导凋亡的情况,进一步了解压疮的病理机制,为有效预防及治疗压疮提供新的靶点及依据。方法根据国内姜丽萍等人研究的大鼠压疮模型建模。将40只雄性Wistar大鼠按照随机数字表方法分为5组,对照组(Control组后称Con组)、受压三个循环组(3circle组,后称3C)、受压六个循环组(6circle组,后称6C组)、受压九个循环组(9circle组,后称9C组)、受压十二个循环(12circle组,后称12C组),每组8只。实验终点时脱颈法处死大鼠并取受压肌肉组织进行检测。HE染色,光镜观察各组受压组织的形态学变化;透射电镜观察各组受压肌肉组织的超微结构的变化;免疫印迹法(Western blot法)检测内质网应激伴侣蛋白78(GRP78)、半胱氨酸天门冬氨酸蛋白酶(caspase-12)的表达情况和线粒体相关蛋白Bax、Bcl-2的表达情况;免疫组织化学法观察并分析GRP78、caspase-12情况。数据采用SPSSl7.0系统进行统计分析,计量资料采用均数±标准差()进行描述,组间差异性比较采用单因素方差分析(one-way ANOVA),以P0.05为有统计学意义。sx?结果1HE染色显示与受压组相比正常对照组,形态正常,肌纤维完整、排列整齐;随着循环周期的延长受压组逐步出现退化的病理变化,表现为肌纤维排列紊乱出现少量的碎片,出现炎症细胞浸润逐渐加重,肌纤维出现溶解,断裂,空泡变性等病理改变。2透射电镜显示对照组肌纤维组织排列整齐紧密,间隙中可见形态正常且完整的线粒体;随循环周期的延长受压组织骨骼肌细胞出现了广泛的线粒体损伤,线粒体排列紊乱,嵴消失,水肿,或成空泡样变性,内质网扩张明显。3Western blot显示受压各组GRP78表达均高于对照组且;而caspase-12与对照组相比只有6C、9C、12C时显著升高,提示循环周期数的增多内质网损伤和其介导的凋亡越严重。同时,能够反映细胞凋亡的Bcl-2/Bax值显示,与对照组相比3C组略有增高,但没有统计学差异;而6C、9C、12C组Bcl-2/Bax值显著减小且呈递减趋势。4免疫组化结果显示在各组中,GRP78的表达与Con组相比3C组,6C组,9C组和12C组的表达均明显增加(P0.05),但6C、9C、12C组的表达较3C组的表达有一定程度的降低且有统计学意义(P0.05)。而caspase-12蛋白的表达,与Con组比较3C组增加不显著,(P0.05);6C组,9C组,12C组与Con组相比均升高明显,且有统计学意义(P0.05)。结论1在大鼠压疮发生过程中激活了内质网应激(GRP78)及其介导的凋亡(caspase-12);2在大鼠压疮发生过程中引起了线粒体的损伤和其介导的凋亡。
[Abstract]:Objective to investigate the mechanism of mitochondria and endoplasmic reticulum stress in the occurrence of pressure sore in rats, the role of mitochondria and endoplasmic reticulum stress and its mediated apoptosis, and to further understand the pathological mechanism of pressure sore. To provide a new target and basis for the effective prevention and treatment of pressure sore. Methods according to Jiang Liping et al. Forty male Wistar rats were randomly divided into five groups: control group (Con group), three circulatory groups (3circle group, 3C) and six circulation groups (6circle group, 6C group). Nine circulation groups (9circle group, 9C group) and twelve circulation groups (12circle group, 12C group) were pressured with 8 rats in each group. At the end of the experiment, the rats were killed by removing neck and the compressed muscle tissue was detected by HE staining. The morphological changes of compressed tissue were observed by light microscope, the ultrastructure of compressed muscle tissue was observed by transmission electron microscope (TEM). The expression of endoplasmic reticulum stress chaperone protein 78 (GRP78), cysteine aspartate protease (caspase-12) and mitochondrial associated protein (Bax,Bcl-2) were detected by Western blotting (Western blot). GRP78,caspase-12 was observed and analyzed by immunohistochemical method. The data were statistically analyzed by SPSSl7.0 system, the measurement data were described by mean 卤standard deviation (卤standard deviation), and the differences between groups were compared by single factor variance analysis (one-way ANOVA), P0.05 was statistically significant). Sx? Results compared with the control group, 1HE staining showed that the shape of the muscle fibers was normal, the muscle fibers were intact and arranged neatly. With the prolongation of the cycle period, the pathological changes appeared gradually in the compression group, which showed that a small amount of fragments appeared in the muscle fiber arrangement disorder, inflammatory cell infiltration was gradually aggravated, the muscle fiber was dissolved and broken. The pathological changes such as vacuolar degeneration. 2 transmission electron microscope showed that the muscle fibers of the control group were arranged neatly and tightly, and normal and complete mitochondria were seen in the interspace. With the prolongation of the cycle, extensive mitochondrial damage appeared in skeletal muscle cells, mitochondria arranged in disorder, ridge disappeared, edema, or vacuolar degeneration. Endoplasmic reticulum dilatation was obvious. 3Western blot showed that the expression of GRP78 in the compression group was higher than that in the control group. Compared with the control group, caspase-12 only increased significantly at 12 C, suggesting that the increase of cycle number and endoplasmic reticulum injury and apoptosis were more serious. At the same time, the Bcl-2/Bax value, which can reflect apoptosis, was slightly higher than that in the control group, but there was no statistical difference. The results of immunohistochemistry showed that the expression of GRP78 was significantly higher in 3C group, 6C group, 9C group and 12C group than in Con group (P0.05), but the expression of GRP78 in 6C group, 6C group and 12C group was significantly higher than that in Con group (P0.05). The expression of 12C group was significantly lower than that of 3C group (P0.05). The expression of caspase-12 protein in 3C group was not significantly higher than that in Con group (P0.05); in 6C group, 9C group, 12C group and Con group, the expression was significantly higher than that in Con group (P0.05). Conclusion (1) Endoplasmic reticulum stress (GRP78) and its mediated apoptosis (caspase-12) were activated in the process of the occurrence of pressure sore in rats, and mitochondrial damage and apoptosis were induced in the process of the occurrence of pressure sore in rats.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R471
[Abstract]:Objective to investigate the mechanism of mitochondria and endoplasmic reticulum stress in the occurrence of pressure sore in rats, the role of mitochondria and endoplasmic reticulum stress and its mediated apoptosis, and to further understand the pathological mechanism of pressure sore. To provide a new target and basis for the effective prevention and treatment of pressure sore. Methods according to Jiang Liping et al. Forty male Wistar rats were randomly divided into five groups: control group (Con group), three circulatory groups (3circle group, 3C) and six circulation groups (6circle group, 6C group). Nine circulation groups (9circle group, 9C group) and twelve circulation groups (12circle group, 12C group) were pressured with 8 rats in each group. At the end of the experiment, the rats were killed by removing neck and the compressed muscle tissue was detected by HE staining. The morphological changes of compressed tissue were observed by light microscope, the ultrastructure of compressed muscle tissue was observed by transmission electron microscope (TEM). The expression of endoplasmic reticulum stress chaperone protein 78 (GRP78), cysteine aspartate protease (caspase-12) and mitochondrial associated protein (Bax,Bcl-2) were detected by Western blotting (Western blot). GRP78,caspase-12 was observed and analyzed by immunohistochemical method. The data were statistically analyzed by SPSSl7.0 system, the measurement data were described by mean 卤standard deviation (卤standard deviation), and the differences between groups were compared by single factor variance analysis (one-way ANOVA), P0.05 was statistically significant). Sx? Results compared with the control group, 1HE staining showed that the shape of the muscle fibers was normal, the muscle fibers were intact and arranged neatly. With the prolongation of the cycle period, the pathological changes appeared gradually in the compression group, which showed that a small amount of fragments appeared in the muscle fiber arrangement disorder, inflammatory cell infiltration was gradually aggravated, the muscle fiber was dissolved and broken. The pathological changes such as vacuolar degeneration. 2 transmission electron microscope showed that the muscle fibers of the control group were arranged neatly and tightly, and normal and complete mitochondria were seen in the interspace. With the prolongation of the cycle, extensive mitochondrial damage appeared in skeletal muscle cells, mitochondria arranged in disorder, ridge disappeared, edema, or vacuolar degeneration. Endoplasmic reticulum dilatation was obvious. 3Western blot showed that the expression of GRP78 in the compression group was higher than that in the control group. Compared with the control group, caspase-12 only increased significantly at 12 C, suggesting that the increase of cycle number and endoplasmic reticulum injury and apoptosis were more serious. At the same time, the Bcl-2/Bax value, which can reflect apoptosis, was slightly higher than that in the control group, but there was no statistical difference. The results of immunohistochemistry showed that the expression of GRP78 was significantly higher in 3C group, 6C group, 9C group and 12C group than in Con group (P0.05), but the expression of GRP78 in 6C group, 6C group and 12C group was significantly higher than that in Con group (P0.05). The expression of 12C group was significantly lower than that of 3C group (P0.05). The expression of caspase-12 protein in 3C group was not significantly higher than that in Con group (P0.05); in 6C group, 9C group, 12C group and Con group, the expression was significantly higher than that in Con group (P0.05). Conclusion (1) Endoplasmic reticulum stress (GRP78) and its mediated apoptosis (caspase-12) were activated in the process of the occurrence of pressure sore in rats, and mitochondrial damage and apoptosis were induced in the process of the occurrence of pressure sore in rats.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R471
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