本文選題：高膽固醇血癥 + 骨質(zhì)疏松　； 參考：《山東大學(xué)》2017年碩士論文

【摘要】：背景和目的:近年來(lái),男性骨質(zhì)疏松患病率和骨質(zhì)疏松相關(guān)性骨折的風(fēng)險(xiǎn)逐年增加,同時(shí)男性高膽固醇血癥的患病率也增加。既往的一些臨床研究關(guān)于高膽固醇血癥和骨密度之間的關(guān)系一直存在爭(zhēng)議,而且大部分的研究都集中于高膽固醇血癥對(duì)女性骨代謝的影響,關(guān)于高膽固醇血癥對(duì)男性骨代謝影響的研究相對(duì)缺乏。本研究本旨在研究高膽固醇血癥對(duì)男性骨代謝的影響并初步探討其作用機(jī)制。方法:以2015年8月-2015年10月在濟(jì)南市翡翠郡社區(qū)流行病學(xué)調(diào)查的216名年齡在18歲以上的男性居民為研究對(duì)象,參與臨床橫斷面研究。血清血脂水平測(cè)定使用的貝克曼化學(xué)分析儀AU5800系統(tǒng)195(貝克曼庫(kù)爾特,東京,日本),血清Beta-CTX、PINP濃度由電化學(xué)發(fā)光免疫測(cè)定(瑞士,cobas 800分析儀)。我們?cè)O(shè)計(jì)兩個(gè)雄性動(dòng)物模型,在高膽固醇飲食模型中,將大鼠分三組,控制組給予正常飲食(NCD,100%標(biāo)準(zhǔn)嚙齒動(dòng)物食物)36周,HCD組給予高膽固醇飲食(HCD)(2%膽固醇和98%標(biāo)準(zhǔn)嚙齒動(dòng)物飼料)36周,誘導(dǎo)外源性高膽固醇血癥,HCD+NCD組給予高膽固醇飲食24周后改為正�？刂骑嬍�12周;在APOE-KO大鼠模型中,給予野生型雄性SD大鼠(Apoe +/+)和雄性Apoe-ko(Apoe-/-)大鼠(先天性高膽固醇血癥模型),喂養(yǎng)標(biāo)準(zhǔn)嚙齒動(dòng)物100%的正常飼料20周。老鼠空腹8小時(shí)被殺,使用自動(dòng)生化分析儀測(cè)量老鼠的血清血脂水平(日本奧林巴,AU5400),使用ELISA試劑盒(優(yōu)爾生,武漢,中國(guó))測(cè)定血清ALPL PINP CTX-Ⅰ和TRAP水平。收集老鼠的左股骨檢測(cè)骨的微觀結(jié)構(gòu)分析和生物力學(xué)分析。從SD大鼠新生幼鼠的頭顱骨提取原代成骨細(xì)胞,用于體外研究。在細(xì)胞實(shí)驗(yàn)中,用不同濃度的膽固醇(美國(guó)Sigma-Aldrich)原代成骨細(xì)胞細(xì)胞12小時(shí),24小時(shí)和48小時(shí)。然后我們用Rt-PCR分析成骨細(xì)胞功能基因ALPL,BGP和Ⅰ型膠原蛋白mRNA表達(dá)水平,用ELISA(優(yōu)爾生,武漢,中國(guó))ALPL,BGP蛋白表達(dá)水平。結(jié)果:與對(duì)照組相比,男性高膽固醇血癥患者骨密度低,骨吸收指標(biāo)(Beta-CTX)和骨形成標(biāo)指標(biāo)(PINP)高;偏相關(guān)分析顯示用年齡,體重指數(shù),糖化血紅蛋白,,FT4,TSH,25(OH)VD和TG校正后,血清膽固醇與BMD呈負(fù)相關(guān),與血清beta-CTX和PINP水平呈正相關(guān);多元線性回歸分析的結(jié)果表明,膽固醇是BMD,beta-CTX和PINP的獨(dú)立影響因子,與BMD呈負(fù)相關(guān),與beta-CTX和PINP呈正相關(guān)。在高膽固醇飲食誘導(dǎo)的外源性高膽固血癥雄性大鼠造模中,與對(duì)照組相比,HCD組大鼠BMD降低,骨吸收指標(biāo)(CTX-I,TRAP)和骨形成指標(biāo)(PINP,BGP和ALPL)升高;HCD損傷了骨微觀結(jié)構(gòu)(骨MicroCT分析),降低了骨強(qiáng)度和韌性(骨生物力學(xué)分析)。撤高膽固醇飲食后,與HCD組比,高膽固醇血癥較低,骨損傷程度較小輕。在APOE-KO誘導(dǎo)的內(nèi)源性高膽固醇血癥雄性大鼠模型中,與WT相比,先天性高膽固醇血癥的Apoe-/-大鼠BMD降低,骨吸收指標(biāo)(CTX-I,TRAP)和骨形成指標(biāo)(PINP,BGP和ALPL)升高;HCD損傷了骨微觀結(jié)構(gòu)(骨MicroCT分析),降低了骨強(qiáng)度和韌性(骨生物力學(xué)分析)。在細(xì)胞實(shí)驗(yàn)中,通過(guò)HE染色,ALP染色,茜素紅和馮庫(kù)薩染色,從新生幼鼠的顱骨提取的原代成骨細(xì)胞為理想的成骨細(xì)胞模型;RT-PCR結(jié)果表明,與對(duì)照組相比,隨著處理的膽固醇水平的增加,ALPL、BGP和一型膠原蛋白mRNA表達(dá)水平逐漸增加。不同時(shí)間用膽固醇處理后,成骨細(xì)胞功能基因mRNA表達(dá)水平?jīng)]有變化;ELISSA結(jié)果表明,與對(duì)照組相比,隨著處理的膽固醇水平的增加,ALPL和BGP蛋白表達(dá)水平逐漸增加。不同時(shí)間用膽固醇處理后,成骨細(xì)胞功能基因mRNA表達(dá)水平?jīng)]有變化。結(jié)論:高膽固醇血癥通過(guò)促進(jìn)破骨細(xì)胞的骨吸收和成骨細(xì)胞的骨形成,提高骨轉(zhuǎn)換率,破壞骨形成和骨吸收之間的平衡,導(dǎo)致骨微觀結(jié)構(gòu)受損害和骨力量和韌性降低,最終可能引起骨質(zhì)疏松和骨質(zhì)疏松相關(guān)性骨折的發(fā)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.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R580;R589.2

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相關(guān)期刊論文 前2條

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本文編號(hào)：2050825