線(xiàn)粒體與內(nèi)質(zhì)網(wǎng)應(yīng)激在壓瘡中的作用機(jī)制
發(fā)布時(shí)間：2018-12-08 12:14
【摘要】：目的探討線(xiàn)粒體與內(nèi)質(zhì)網(wǎng)應(yīng)激在大鼠壓瘡發(fā)生中的作用機(jī)制,線(xiàn)粒體及內(nèi)質(zhì)網(wǎng)應(yīng)激的作用及其介導(dǎo)凋亡的情況,進(jìn)一步了解壓瘡的病理機(jī)制,為有效預(yù)防及治療壓瘡提供新的靶點(diǎn)及依據(jù)。方法根據(jù)國(guó)內(nèi)姜麗萍等人研究的大鼠壓瘡模型建模。將40只雄性Wistar大鼠按照隨機(jī)數(shù)字表方法分為5組,對(duì)照組(Control組后稱(chēng)Con組)、受壓三個(gè)循環(huán)組(3circle組,后稱(chēng)3C)、受壓六個(gè)循環(huán)組(6circle組,后稱(chēng)6C組)、受壓九個(gè)循環(huán)組(9circle組,后稱(chēng)9C組)、受壓十二個(gè)循環(huán)(12circle組,后稱(chēng)12C組),每組8只。實(shí)驗(yàn)終點(diǎn)時(shí)脫頸法處死大鼠并取受壓肌肉組織進(jìn)行檢測(cè)。HE染色,光鏡觀(guān)察各組受壓組織的形態(tài)學(xué)變化;透射電鏡觀(guān)察各組受壓肌肉組織的超微結(jié)構(gòu)的變化;免疫印跡法(Western blot法)檢測(cè)內(nèi)質(zhì)網(wǎng)應(yīng)激伴侶蛋白78(GRP78)、半胱氨酸天門(mén)冬氨酸蛋白酶(caspase-12)的表達(dá)情況和線(xiàn)粒體相關(guān)蛋白Bax、Bcl-2的表達(dá)情況;免疫組織化學(xué)法觀(guān)察并分析GRP78、caspase-12情況。數(shù)據(jù)采用SPSSl7.0系統(tǒng)進(jìn)行統(tǒng)計(jì)分析,計(jì)量資料采用均數(shù)±標(biāo)準(zhǔn)差()進(jìn)行描述,組間差異性比較采用單因素方差分析(one-way ANOVA),以P0.05為有統(tǒng)計(jì)學(xué)意義。sx?結(jié)果1HE染色顯示與受壓組相比正常對(duì)照組,形態(tài)正常,肌纖維完整、排列整齊;隨著循環(huán)周期的延長(zhǎng)受壓組逐步出現(xiàn)退化的病理變化,表現(xiàn)為肌纖維排列紊亂出現(xiàn)少量的碎片,出現(xiàn)炎癥細(xì)胞浸潤(rùn)逐漸加重,肌纖維出現(xiàn)溶解,斷裂,空泡變性等病理改變。2透射電鏡顯示對(duì)照組肌纖維組織排列整齊緊密,間隙中可見(jiàn)形態(tài)正常且完整的線(xiàn)粒體;隨循環(huán)周期的延長(zhǎng)受壓組織骨骼肌細(xì)胞出現(xiàn)了廣泛的線(xiàn)粒體損傷,線(xiàn)粒體排列紊亂,嵴消失,水腫,或成空泡樣變性,內(nèi)質(zhì)網(wǎng)擴(kuò)張明顯。3Western blot顯示受壓各組GRP78表達(dá)均高于對(duì)照組且;而caspase-12與對(duì)照組相比只有6C、9C、12C時(shí)顯著升高,提示循環(huán)周期數(shù)的增多內(nèi)質(zhì)網(wǎng)損傷和其介導(dǎo)的凋亡越嚴(yán)重。同時(shí),能夠反映細(xì)胞凋亡的Bcl-2/Bax值顯示,與對(duì)照組相比3C組略有增高,但沒(méi)有統(tǒng)計(jì)學(xué)差異;而6C、9C、12C組Bcl-2/Bax值顯著減小且呈遞減趨勢(shì)。4免疫組化結(jié)果顯示在各組中,GRP78的表達(dá)與Con組相比3C組,6C組,9C組和12C組的表達(dá)均明顯增加(P0.05),但6C、9C、12C組的表達(dá)較3C組的表達(dá)有一定程度的降低且有統(tǒng)計(jì)學(xué)意義(P0.05)。而caspase-12蛋白的表達(dá),與Con組比較3C組增加不顯著,(P0.05);6C組,9C組,12C組與Con組相比均升高明顯,且有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論1在大鼠壓瘡發(fā)生過(guò)程中激活了內(nèi)質(zhì)網(wǎng)應(yīng)激(GRP78)及其介導(dǎo)的凋亡(caspase-12);2在大鼠壓瘡發(fā)生過(guò)程中引起了線(xiàn)粒體的損傷和其介導(dǎo)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 鹵standard deviation (鹵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.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：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 鹵standard deviation (鹵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.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R471
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