快速衰老小鼠张口呼吸模型制备及其线粒体功能损伤研究

发布时间：2017-12-28 17:38

本文关键词：快速衰老小鼠张口呼吸模型制备及其线粒体功能损伤研究　出处：《江苏大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的1.探索安全高效制备张口呼吸小鼠模型方法。2.制备基于快速衰老小鼠(SAMP)的张口呼吸模型,并探究张口呼吸对上气道组织线粒体功能损伤的影响。方法1.将20只C57BL/6分为缝合组(n=10)与填塞组(n=10),分别行外鼻孔缝合与鼻腔填塞,比较分析外鼻孔缝合与鼻腔填塞两种造模方法下小鼠死亡率与鼻腔阻塞物脱落率,选择低死亡率低脱落率的鼻腔阻塞方法用于进一步造模研究。2.快速衰老小鼠张口呼吸模型线粒体功能损伤研究:将4月龄SAMP6小鼠12只与SAMR1小鼠12只分别行鼻腔阻塞,制备张口呼吸模型(SAMP6_OR和SAMR1_OR)。造模后,各取4只备用于1周后取材。剩余小鼠分别饲养至6月龄(n=4)、8月龄(n=4),取相同数量同月龄SAMP6小鼠与SAMR1小鼠与之匹配备取材。每只小鼠取皮肤、上气道肌肉、腹直肌、上气道黏膜、颏舌肌、臀大肌、白细胞制备细胞样本,加载JC-1并进行线粒体膜电位检测。比较分析4、6、8月龄SAMP6与SAMR1、SAMP6_OR与SAMP6、SAMR1_OR与SAMR1小鼠各组织线粒体膜电位。分析4、6、8月龄SAMP6与SAMR1小鼠各组织线粒体膜电位变化趋势。结果1.缝合造模法与填塞造模法造模死亡率与鼻腔阻塞物脱落率比较1)鼻腔缝合法造模死亡率(70%)显著高于鼻腔填塞法(0%)(P0.05)。2)缝合组线结脱落率(50%)与鼻腔填塞组填塞物脱落率(20%)无显著差异(P0.05)。2.快速衰老小鼠张口呼吸模型线粒体功能损伤研究1)SAMP6与SAMR1小鼠相比,全身大部分组织线粒体膜电位显著降低(P0.05)。2)SAMP6小鼠4、6、8月龄之间皮肤(P0.05)、上气道肌肉(P0.05)、腹直肌(P0.05)、上气道黏膜(P0.05)、颏舌肌(P0.05)、臀大肌(P0.05)、白细胞(P0.05)线粒体膜电位均存在显著差异。3)SAMR1小鼠4、6、8月龄之间上气道黏膜(P0.05)、颏舌肌(P0.05)、白细胞(P0.05)线粒体膜电位之间存在显著差异。但皮肤(P0.05)、上气道肌肉(P0.05)、腹直肌(P0.05)、臀大肌(P0.05)线粒体膜电位无显著差异。4)SAMP6_OR小鼠皮肤、腹直肌、上气道肌肉与黏膜组织、白细胞线粒体膜电位显著低于SAMP6小鼠(P0.05)。5)SAMPR1_OR小鼠上气道肌肉线粒体膜电位、白细胞膜电位显著低于SAMPR1小鼠(P0.05)。结论1.使用鼻腔填塞法进行张口呼吸小鼠模型造模创伤小,死亡率低,不易脱落,可用于进一步造模研究。2.快速衰老与自然衰老过程中线粒体功能均随年龄增长而下降,各组织衰老进程存在差异。快速衰老小鼠张口呼吸模型上气道肌肉与黏膜损伤更为严重,在此基础上还出现了皮肤、腹直肌、白细胞线粒体功能损伤,这种损伤存在全身性趋势。但自然衰老小鼠张口呼吸模型中仅上气道肌肉、白细胞损伤较为明显,是否存在全身性趋势有待进一步研究。
[Abstract]:Objective 1. to explore a safe and efficient method of making mouth breathing mice model. 2. to prepare the mouth breathing model based on rapid aging mice (SAMP) and to explore the effect of mouth breathing on mitochondrial dysfunction in upper airway tissue. Methods 1. 20 C57BL/6 divided into suture group (n=10) and packing group (n=10), respectively. The external naris suture and nasal packing, a comparative analysis of the external naris suture and nasal packing two models under the mortality of mice with nasal obstruction abscission rate, low mortality low shedding rate of nasal obstruction for further modeling research. 2., fast aging mice mouth breathing model mitochondrial function damage research: 4 month old SAMP6 mice and 12 SAMR1 mice 12 nasal blockage, to prepare mouth breathing model (SAMP6_OR and SAMR1_OR). After the model, 4 were taken for 1 weeks. The remaining mice were fed to 6 month old (n=4) and 8 month old (n=4) respectively, and the same number of SAMP6 mice of the same month age and SAMR1 mice were obtained. Each mouse was prepared from skin, upper airway muscle, rectus abdominis, upper airway mucosa, genu lingual muscle, gluteus maximus and white blood cell. Samples were loaded with JC-1 and mitochondrial membrane potential was detected. The mitochondrial membrane potential of 4, 6, 8 month old SAMP6 and SAMR1, SAMP6_OR and SAMP6, SAMR1_OR and SAMR1 mice was compared and analyzed. The changes of mitochondrial membrane potential in the tissues of 4, 6, 8 month old SAMP6 and SAMR1 mice were analyzed. Results 1. there was a 1 Comparison between the rate of die building and the rate of obstruction of the nasal cavity by suturing and modeling. 1) the mortality rate of nasal cavity suture was higher than that of nasal packing (0%) (P0.05). 2) there was no significant difference between the loss rate of line knot (50%) in the suture group and the loss rate of the stuffing in the nasal packing group (20%) (P0.05). 2., fast aging mice mouth breathing model mitochondrial dysfunction. 1) SAMP6, compared with SAMR1 mice, the mitochondrial membrane potential of most tissues decreased significantly (P0.05). 2) there were significant differences in mitochondrial membrane potential between skin, P0.05, upper airway muscle (P0.05), rectus abdominis muscle (P0.05), upper airway mucosa (P0.05), genu gluteal muscle (P0.05), gluteus maximus muscle (P0.05) and white blood cell (P0.05) in SAMP6 mice between 4, 6 and 8 month old. 3) there were significant differences in the mitochondrial membrane potential of the upper airway mucosa (P0.05), genial tongue muscle (P0.05) and leukocyte (P0.05) between 4, 6 and 8 month old of SAMR1 mice. But there was no significant difference in the mitochondrial membrane potential of the skin (P0.05), the upper airway muscle (P0.05), the rectus abdominis (P0.05) and the gluteus maximus (P0.05). 4) the potential of SAMP6_OR mice skin, rectus abdominis muscle, upper airway muscle and mucous membrane tissue and leukocyte mitochondrial membrane potential were significantly lower than that of SAMP6 mice (P0.05). 5) the mitochondrial membrane potential and the white cell membrane potential of the upper airway muscle of SAMPR1_OR mice were significantly lower than that of SAMPR1 mice (P0.05). Conclusion 1. using the nasal cavity filling method to make the mouth breathing mice model is less traumatic, the death rate is low, and it is not easy to fall off. It can be used for further study of the model. 2. in the process of rapid senescence and natural aging, the function of mitochondria decreased with age, and there was a difference in the aging process of each tissue. In the fast aging mice, the upper airway muscle and mucosa injury is more serious, and there are skin damage, rectus abdominis and white cell mitochondrial dysfunction. However, only the upper airway muscle and white blood cell damage in the mouth breathing model of natural aging mice are more obvious, and whether there is a systemic trend is to be further studied.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R766;R-332

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