高胆固醇血症对男性骨代谢的影响及其机制的研究

发布时间：2018-06-22 01:00

本文选题：高胆固醇血症 + 骨质疏松　；参考：《山东大学》2017年硕士论文

【摘要】：背景和目的:近年来,男性骨质疏松患病率和骨质疏松相关性骨折的风险逐年增加,同时男性高胆固醇血症的患病率也增加。既往的一些临床研究关于高胆固醇血症和骨密度之间的关系一直存在争议,而且大部分的研究都集中于高胆固醇血症对女性骨代谢的影响,关于高胆固醇血症对男性骨代谢影响的研究相对缺乏。本研究本旨在研究高胆固醇血症对男性骨代谢的影响并初步探讨其作用机制。方法:以2015年8月-2015年10月在济南市翡翠郡社区流行病学调查的216名年龄在18岁以上的男性居民为研究对象,参与临床横断面研究。血清血脂水平测定使用的贝克曼化学分析仪AU5800系统195(贝克曼库尔特,东京,日本),血清Beta-CTX、PINP浓度由电化学发光免疫测定(瑞士,cobas 800分析仪)。我们设计两个雄性动物模型,在高胆固醇饮食模型中,将大鼠分三组,控制组给予正常饮食(NCD,100%标准啮齿动物食物)36周,HCD组给予高胆固醇饮食(HCD)(2%胆固醇和98%标准啮齿动物饲料)36周,诱导外源性高胆固醇血症,HCD+NCD组给予高胆固醇饮食24周后改为正常控制饮食12周;在APOE-KO大鼠模型中,给予野生型雄性SD大鼠(Apoe +/+)和雄性Apoe-ko(Apoe-/-)大鼠(先天性高胆固醇血症模型),喂养标准啮齿动物100%的正常饲料20周。老鼠空腹8小时被杀,使用自动生化分析仪测量老鼠的血清血脂水平(日本奥林巴,AU5400),使用ELISA试剂盒(优尔生,武汉,中国)测定血清ALPL PINP CTX-Ⅰ和TRAP水平。收集老鼠的左股骨检测骨的微观结构分析和生物力学分析。从SD大鼠新生幼鼠的头颅骨提取原代成骨细胞,用于体外研究。在细胞实验中,用不同浓度的胆固醇(美国Sigma-Aldrich)原代成骨细胞细胞12小时,24小时和48小时。然后我们用Rt-PCR分析成骨细胞功能基因ALPL,BGP和Ⅰ型胶原蛋白mRNA表达水平,用ELISA(优尔生,武汉,中国)ALPL,BGP蛋白表达水平。结果:与对照组相比,男性高胆固醇血症患者骨密度低,骨吸收指标(Beta-CTX)和骨形成标指标(PINP)高;偏相关分析显示用年龄,体重指数,糖化血红蛋白,,FT4,TSH,25(OH)VD和TG校正后,血清胆固醇与BMD呈负相关,与血清beta-CTX和PINP水平呈正相关;多元线性回归分析的结果表明,胆固醇是BMD,beta-CTX和PINP的独立影响因子,与BMD呈负相关,与beta-CTX和PINP呈正相关。在高胆固醇饮食诱导的外源性高胆固血症雄性大鼠造模中,与对照组相比,HCD组大鼠BMD降低,骨吸收指标(CTX-I,TRAP)和骨形成指标(PINP,BGP和ALPL)升高;HCD损伤了骨微观结构(骨MicroCT分析),降低了骨强度和韧性(骨生物力学分析)。撤高胆固醇饮食后,与HCD组比,高胆固醇血症较低,骨损伤程度较小轻。在APOE-KO诱导的内源性高胆固醇血症雄性大鼠模型中,与WT相比,先天性高胆固醇血症的Apoe-/-大鼠BMD降低,骨吸收指标(CTX-I,TRAP)和骨形成指标(PINP,BGP和ALPL)升高;HCD损伤了骨微观结构(骨MicroCT分析),降低了骨强度和韧性(骨生物力学分析)。在细胞实验中,通过HE染色,ALP染色,茜素红和冯库萨染色,从新生幼鼠的颅骨提取的原代成骨细胞为理想的成骨细胞模型;RT-PCR结果表明,与对照组相比,随着处理的胆固醇水平的增加,ALPL、BGP和一型胶原蛋白mRNA表达水平逐渐增加。不同时间用胆固醇处理后,成骨细胞功能基因mRNA表达水平没有变化;ELISSA结果表明,与对照组相比,随着处理的胆固醇水平的增加,ALPL和BGP蛋白表达水平逐渐增加。不同时间用胆固醇处理后,成骨细胞功能基因mRNA表达水平没有变化。结论:高胆固醇血症通过促进破骨细胞的骨吸收和成骨细胞的骨形成,提高骨转换率,破坏骨形成和骨吸收之间的平衡,导致骨微观结构受损害和骨力量和韧性降低,最终可能引起骨质疏松和骨质疏松相关性骨折的发生。
[Abstract]:Background and purpose: in recent years, the risk of osteoporosis associated with osteoporosis in men is increasing year by year, and the prevalence of male hypercholesterolemia is also increasing. Some previous clinical studies have been disputed about the relationship between hypercholesterolemia and bone density, and most of the studies are focused on high cholesterol. The effect of alcoholemia on bone metabolism in women and the relative lack of study on the effect of hypercholesterolemia on male bone metabolism. This study was designed to study the effect of hypercholesterolemia on male bone metabolism and to explore its mechanism. Methods: 216 years in the 216 year of the August 2015 -2015 year's community epidemiological survey in the jadeite County in October. AU5800 system 195 (Beckman Kurt, Tokyo, Japan), serum Beta-CTX, PINP concentration was determined by electrochemiluminescence immunoassay (Switzerland, Cobas 800 analyzer). We designed two male animal models. The serum lipid level was measured by the Backman chemical analyzer (Backman, Tokyo, Japan). In the high cholesterol diet model, the rats were divided into three groups. The control group was given a normal diet (NCD, 100% standard rodent food) for 36 weeks. The HCD group was given a high cholesterol diet (HCD) (2% cholesterol and 98% standard rodent feed) for 36 weeks, induced exogenous hypercholesterolemia, and the group HCD+NCD was given a high cholesterol diet for 24 weeks to normal control. Diet for 12 weeks; in the APOE-KO rat model, wild type male SD rats (Apoe + / +) and male Apoe-ko (Apoe-/-) rats (congenital hypercholesterolemia model) were fed with normal feed of standard rodents for 20 weeks. The rats were killed for 8 hours on an empty stomach. The serum lipid levels of mice were measured by automatic biochemistry analyzer (Japanese orange, AU5). 400) the serum ALPL PINP CTX- I and TRAP levels were measured with the ELISA Kit (ulson, Wuhan, China). The microstructural analysis and biomechanical analysis of the mouse's left femur bone were collected. The primary osteoblasts were extracted from the skull bone of the newborn rats of SD and used for the study in vitro. In the cell experiment, the different concentrations of cholesterol (beauty) were used in the cell experiment. Sigma-Aldrich) primary osteoblast cells for 12 hours, 24 hours and 48 hours. Then we used Rt-PCR to analyze the expression level of osteoblast function gene ALPL, BGP and type I collagen mRNA, ALPL, BGP protein expression level with ELISA (Olson, Wuhan, China). Results: compared with the control group, the bone density of male hypercholesterolemic patients was low, bone density, bone and bone density. Absorption index (Beta-CTX) and bone formation standard index (PINP) were high. Partial correlation analysis showed that serum cholesterol and BMD were negatively correlated with age, body mass index, glycosylated hemoglobin, FT4, TSH, 25 (OH) VD and TG, and positive correlation with serum beta-CTX and PINP levels. The results of multivariate linear regression analysis showed that cholesterol was BMD, beta-CTX, and PINP. Independent influence factors were negatively correlated with BMD and positively correlated with beta-CTX and PINP. In the model of exogenous hypercholesterolemia male rats induced by high cholesterol diet, compared with the control group, BMD decreased in the HCD group, the index of bone absorption (CTX-I, TRAP) and bone formation index (PINP, BGP and ALPL) increased; HCD damaged the bone microstructure (bone MicroCT analysis). Lower bone strength and toughness (bone biomechanical analysis). After a high cholesterol diet, the hypercholesterolemia was lower and the bone damage was smaller than the HCD group. In the male rat model of endogenous hypercholesterolemia induced by APOE-KO, the BMD of the Apoe-/- rats with congenital hypercholesterolemia was reduced, and the index of bone absorption (CTX-I, TRA) was compared with that of WT. P) and bone formation index (PINP, BGP and ALPL) increased; HCD damaged bone microstructure (bone MicroCT analysis), reduced bone strength and toughness (bone biomechanical analysis). In cell experiments, HE staining, ALP staining, alizarin red and von cuosa staining, the original osteoblasts extracted from the skull of newborn rats were ideal osteoblast models; RT-P CR results showed that compared with the control group, the expression level of ALPL, BGP and type 1 collagen mRNA increased gradually with the increase of cholesterol level in the control group. The expression level of osteoblast functional gene mRNA was not changed after the treatment with cholesterol at different time. The ELISSA results showed that compared with the control group, the cholesterol level increased, AL The expression level of PL and BGP protein increased gradually. The expression level of osteoblast functional gene mRNA was not changed at different time. Conclusion: Hypercholesterolemia can increase the rate of bone transformation by promoting bone absorption of osteoclast and bone formation of osteoblast, and destroy the balance between bone formation and bone absorption, resulting in bone microstructure. Damage and bone strength and toughness may eventually lead to osteoporosis and osteoporosis related fractures.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R580;R589.2

【参考文献】