乳铁蛋白对去卵巢大鼠IGF-IR、IGFBP-2、IGFBP-4mRNA表达影响的研究

发布时间：2018-04-13 20:28

本文选题：乳铁蛋白 + 骨质疏松　；参考：《福建医科大学》2015年硕士论文

【摘要】：目的:本研究旨在观察乳铁蛋白(Lactoferrin LF)对去卵巢大鼠IGF-IR/IGFBP-2/IGFBP-4m RNA基因表达的影响,探讨乳铁蛋白是否通过介导IGF-IR/IGFBP-2/IGFBP-4m RNA基因表达机制治疗绝经后骨质疏松。方法:选取6个月大小,体重约在270g左右,雌性SD(Sprague-Dawley)大鼠70只,适应性喂养1周后,入组大鼠随机分为去卵巢模型组(Ovx)60只与假手术组(Sham)10只,把去卵巢大鼠模型组再次分组,随机分为不同干预治疗组6组,骨质疏松模型组(Con组)10只,实验中作为对照组:[蒸馏水2ml/(只·天)灌胃],牛血清白蛋白组(BSA组)10只:[100mg/kg/d+蒸馏水2ml,灌胃],不同浓度剂量的乳铁蛋白治疗组LF1-3(LF1、LF2和LF3组各10只):[0.1g/(kg·d)+蒸馏水2ml,1g/(kg·d)+蒸馏水2ml,2g/(kg·d)+蒸馏水2ml,灌胃],雌激素治疗组(E2组)10只[0.1 mg/(kg·周),肌肉注射]。连续干预治疗6个月后,通过观察乳铁蛋白干预前后,体重有无明显差异;通过制作HE石蜡病理切片,使用显微镜观察骨组织结构变化。再通过Real Time RT-PCR法即实时荧光定量聚合酶链反应,检测去卵巢大鼠骨组织IGF-1R、IGFBP-2、IGFBP-4m RNA表达的改变。结果:1.乳铁蛋白干预前,各组大鼠体重均无明显差异,均P0.05,大鼠去卵巢手术后,以各种方法干预治疗6个月后,除了E2组外,其余各组大鼠体重均明显升高,均P0.05,提示差别有统计学意义,体重增加最多的组为Con组,Con组大鼠均明显重于Sham组和E2组,均P0.05,提示差别有统计学意义,以不同浓度乳铁蛋白进行干预的LF1-3组及牛血清白蛋白组大鼠体重与Con组相比,虽然均比Con组体重低,但均P0.05,提示无统计学意义。2.骨组织HE染色病理切片显示:Sham组、E2组骨小梁排列密集、饱满,间距小,细胞数量多;而Con组、BSA组骨小梁变细、断裂,骨小梁距离增宽,排列稀疏,细胞数量少,骨小梁结构出现较大的空白区域;而乳铁蛋白灌胃组LF1-3介于二者之间,且骨小梁排列紧密程度LF3好于LF2,LF2好于LF1。表明成功建立去卵巢大鼠骨质疏松模型。3.实时荧光定量PCR结果显示:Con组IGFBP-2、IGFBP-4 m RNA相对于Sham组,相对表达量均显著增高,均P0.01,差别有显著统计学意义;相对于Sham组,Con组IGF-1Rm RNA相对表达量明显降低,均P0.01,差别有显著统计学意义;与Con组比较LF1-3组(100 mg/kg,1 g/kg and 2 g/kg)与E2组IGFBP-2、IGFBP-4m RNA相对表达量明显下降,且随着乳铁蛋白浓度的升高下降得越明显,均P0.01,差别有显著统计学意义;而LF2-3组(1 g/kg and 2 g/kg)与E2组IGF-1R m RNA相对表达量均显著高于Con组,且随着乳铁蛋白浓度的升高上升得越明显,均P0.05,差别有统计学意义,牛血清白蛋白BSA组(100mg/kg/d)IGF-1R、IGFBP-2、IGFBP-4m RNA相对表达量相对于Con组基本不变,均P0.05,两者之间无统计学意义。结论:1.成功建立去卵巢大鼠骨质疏松模型。2.乳铁蛋白可能通过抑制IGFBP-2、IGFBP-4 m RNA水平,促进IGF-1R m RNA水平的表达来改善骨质疏松。
[Abstract]:Objective: the purpose of this study was to observe the effects of lactoferrin (Lactoferrin LF) to IGF-IR/IGFBP-2/IGFBP-4m RNA gene expression in ovariectomized rats, to investigate whether lactoferrin mediated by IGF-IR/IGFBP-2/IGFBP-4m RNA gene expression mechanism for treatment of postmenopausal osteoporosis. Methods: a total of 6 months, weight about 270g, female SD (Sprague-Dawley) 70 rats. After 1 weeks of feeding, the rats were randomly divided into ovariectomized group (Ovx) 60 rats with sham operation group (Sham) 10, the ovariectomized rat model group again group, were randomly divided into intervention group with 6 group, osteoporosis model group (Con group) 10. In the experiment as the control group: distilled water 2ml/ (only day) by gavage, bovine serum albumin group (group BSA) 10: [100mg/kg/d+ 2ml of distilled water by gavage, different doses of lactoferrin in treatment group LF1-3 (LF1, LF2 and LF3 group with 10 rats in each group: [0.1g/ (kg - D)) + steamed The distilled water 2ml, 1g/ (kg - D) + distilled water 2ml, 2g/ (kg - D) + 2ml of distilled water by gavage, estrogen treatment group (group E2) 10 [0.1 mg/ (kg), intramuscular injection of continuous intervention. After 6 months of treatment, observed before and after intervention by lactoferrin. There is no obvious difference in weight; through the production of HE paraffin sections, using microscope to observe the microstructure changes of bone. Then Real Time RT-PCR real-time fluorescence quantitative polymerase chain reaction, detection of bone tissue of ovariectomized rats IGF-1R, IGFBP-2, IGFBP-4m RNA expression changes. Results: 1. lactoferrin before intervention showed no significant difference., the body weight of the rats were P0.05 in ovariectomized rats after surgery, with various methods of intervention after 6 months of treatment, except for E2 group, body weight of the rats were significantly increased, P0.05, suggesting that there was a statistically significant difference, the largest increase in weight of group Con, Con group rats were significantly heavier in the Sham group and E2 group, P0.05, suggesting that there was a statistically significant difference, LF1-3 group and bovine serum albumin in rats compared with Con group with different concentrations of lactoferrin intervention, although low body weight was higher than Con group, but P0.05, showed no statistically significant.2. HE staining of bone tissue pathology showed: Sham group, E2 group of trabecular bone densely arranged, plump, small spacing, number of cells; and Con group, BSA group, trabecular bone became thin, fracture, bone trabecula widened the distance, sparse, small numbers of cells, bone trabecula appeared large blank area; between lactoferrin gavage group LF1-3 between the two, and the small bone the beam of closely spaced degree of LF3 is better than LF2, LF2 in LF1. showed that the successful establishment of osteoporosis in ovariectomized rats model of.3. real-time fluorescence quantitative PCR showed that: Con group IGFBP-2, IGFBP-4 m RNA compared with Sham group, the relative expression of P0.01 were significantly increased, there is a significant difference, statistically significant Yi; compared with Sham group, Con group, IGF-1Rm RNA expression was obviously decreased, P0.01, the difference was significant; compared with group Con and group LF1-3 (100 mg/kg, 1 g/kg and 2 g/kg) and IGFBP-2 E2 group, IGFBP-4m RNA expression was obviously decreased, and increased with the increasing concentration of lactoferrin decreased more obviously that was P0.01, the difference was significant; and group LF2-3 (1 g/kg and 2 g/kg) and the relative expression of E2 IGF-1R m in RNA group were significantly higher than Con group, and with the increase of lactoferrin concentration increased more obviously, P0.05, the difference was statistically significant, bovine serum albumin group (100mg/kg/d BSA) IGF-1R, IGFBP-2, the relative expression compared with Con group P0.05 IGFBP-4m RNA were basically unchanged, and no statistical significance between the two. Conclusion: 1. successfully established osteoporosis in ovariectomized rats model of.2. lactoferrin possibly through inhibition of IGFBP-2, IGFBP-4 m RNA, pro The expression of IGF-1R m RNA levels to improve osteoporosis.

【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R580

【参考文献】