本文選題：熊果酸 + 酒精性骨質(zhì)疏松��； 參考：《青島大學》2017年碩士論文

【摘要】：目的:通過觀察熊果酸對酒精性骨質(zhì)疏松大鼠股骨骨密度(BMD)、骨鈣素(BGP)、骨形成蛋白-2(BMP-2)、血清鈣、磷及炎性細胞因子TNF-α和IL-1β等的影響,初步探討熊果酸對酒精所致骨質(zhì)疏松大鼠的作用及可能機制,并分析熊果酸對酒精性骨質(zhì)疏松大鼠腸道菌群多樣性的影響。方法:1.動物分組及給藥:2月齡SPF級雄性Wistar大鼠90只,大鼠適應性喂養(yǎng)7 d后,按體重隨機分為6組(分別為:正常對照組、熊果酸對照組、酒精模型組、熊果酸低、中、高劑量組),每組15只,分籠喂養(yǎng),每日自由進水,進食。酒精模型組大鼠每日給予50%(V/V)酒精8 m L/kg·bw·d,灌胃2周后,劑量提升至12 m L/kg·bw·d,繼續(xù)灌胃6周;正常對照組每日灌胃給予生理鹽水;熊果酸對照組每日給予150 mg/kg·bw·d熊果酸;熊果酸低、中、高劑量組每日分別灌胃給予50、100、150 mg/kg·bw·d熊果酸,酒精劑量同模型組;實驗共持續(xù)8周,并記錄大鼠每周體重,以調(diào)整酒精及熊果酸灌胃量。末次灌胃給藥后禁食不禁水12小時,3%戊巴比妥鈉麻醉,腹主動脈取血,剝離股骨固定。2.股骨病理學觀察:HE染色觀察大鼠股骨的病理學變化。3.股骨骨密度(BMD)測定:DEXA骨密度儀檢測股骨骨密度。4.采用酶聯(lián)免疫吸附法(ELISA)檢測大鼠血清骨鈣素(BGP)、骨形成蛋白-2(BMP-2)濃度的水平。5.采用酶聯(lián)免疫吸附法(ELISA)檢測大鼠血清炎性細胞因子腫瘤壞死因子-α(TNF-α)和白介素-β(IL-1β)含量6.比色法測定血清鈣(Ca)濃度;磷鉬酸法測定血清磷(P)濃度。7.采用DGGE-PCR技術對糞便腸道菌群進行指紋圖譜聚類性分析。結(jié)果:1.HE染色觀察結(jié)果:正常對照組大鼠骨小梁致密、規(guī)則且較粗,粗細均勻;而酒精模型組大鼠骨小梁稀松、不規(guī)則、粗細不均勻,甚至可見骨小梁斷裂;熊果酸中、高劑量組與酒精模型組大鼠相比較,骨小梁致密、規(guī)則、較厚、粗細均勻,未見骨小梁斷裂;熊果酸低劑量組大鼠改善不明顯。2.股骨骨密度:與正常對照組相比較,酒精模型組大鼠的股骨BMD明顯降低,且差異具有統(tǒng)計學意義(P0.05)。但經(jīng)8周熊果酸干預后,可見除熊果酸低劑量組外,熊果酸中、高劑量組大鼠股骨BMD均明顯升高,與酒精模型組相比,均具有統(tǒng)計學差異(P0.05)。3.血清BGP、BMP-2結(jié)果:酒精模型組與正常對照組相比,BGP含量明顯降低,且差異具有統(tǒng)計學意義(P0.05);熊果酸中、高劑量組與酒精模型組相比較,BGP含量均有所升高,且差異有統(tǒng)計學意義(P0.05),但熊果酸低劑量組與酒精模型組相比無統(tǒng)計學差異,僅具有數(shù)值上的升高。酒精模型組與正常對照組相比,可見模型組大鼠血清BMP-2含量明顯降低,且差異具有統(tǒng)計學意義(P0.05);而熊果酸高劑量組與酒精模型組相比,血清BMP-2含量明顯升高,且有統(tǒng)計學差異(P0.05),但熊果酸低、中劑量組血清BMP-2水平變化不大,與酒精模型組相比無統(tǒng)計學差異。4.血清Ca、P結(jié)果:酒精模型組大鼠血清Ca、P水平均有明顯降低,與正常對照組相比較,差異有統(tǒng)計學意義(P0.05);與酒精模型組相比較,熊果酸中、高劑量組血清Ca和P含量明顯回升,且均具有統(tǒng)計學差異(P0.05),但熊果酸低劑量組Ca、P含量雖有數(shù)值上的增加,但無統(tǒng)計學差異。5.血清TNF-α和IL-1β結(jié)果:相對于正常對照組,酒精模型組TNF-α和IL-1β的水平明顯升高,差異具有統(tǒng)計學意義(P0.05);而熊果酸干預后顯著抑制了炎性因子的升高,與酒精模型組相比較,TNF-α和IL-1β的水平顯著下降,其中熊果酸高劑量組作用明顯,且具有統(tǒng)計學差異(P0.05)。6.腸道菌群DGGE-PCR指紋圖譜分析:酒精模型組大鼠腸道菌群多樣性明顯減少且與正常組的相似性較低;而熊果酸高劑量組大鼠腸道菌群結(jié)構更加接近于正常組。結(jié)論:1.通過對大鼠股骨骨密度測定及股骨病理學觀察,初步證實熊果酸對酒精所致的大鼠骨質(zhì)疏松具有顯著的改善作用。2.熊果酸對酒精性骨質(zhì)疏松的保護機制可能與其能夠促進骨形成,降低骨吸收,抑制骨礦物質(zhì)的流失及炎性細胞因子過表達有關。3.熊果酸能夠調(diào)節(jié)因酒精攝入導致的腸道菌群多樣性減少,提示其改善酒精性骨質(zhì)疏松的作用可能與其對腸道菌群的調(diào)節(jié)作用有關。
[Abstract]:Objective: To investigate the effect of ursolic acid on femur bone density (BMD), osteocalcin (BGP), bone morphogenetic protein -2 (BMP-2), serum calcium, phosphorus, and inflammatory cytokine TNF- alpha and IL-1 beta in rats with alcoholic osteoporosis, and to investigate the effect and possible mechanism of ursolic acid on alcohol induced osteoporosis rats and to analyze the effect of ursolic acid on alcohol induced osteoporosis. Methods: the effect of the diversity of intestinal microflora in rats. Methods: 1. animal groups and drug delivery: 2 month old SPF male Wistar rats, 90 rats were fed for 7 d. The rats were randomly divided into 6 groups according to their weight (the normal control group, ursolic acid control group, alcohol model group, ursolic acid low, middle, high dose group), 15 rats in each group, fed by cage and free intake of water daily, The rats in the alcohol model group were given 50% (V/V) alcohol 8 m L/kg. BW. D every day. After 2 weeks of gavage, the dosage was increased to 12 m L/kg. BW. D and continued for 6 weeks. The normal control group was given the normal saline daily; the ursolic acid control group was given 150 mg/kg. BW D ursolic acid daily; the ursolic acid was low, middle, and high dose groups were given 50100150 daily to 50100150. Mg/kg. BW. D ursolic acid, alcohol dose and model group; the experiment lasted for 8 weeks, and the rats' weekly weight was recorded in order to adjust alcohol and ursolic acid perfusion. After the last irrigation, the fasting was 12 hours, 3% pentobarbital sodium anaesthesia, the abdominal aorta took blood, and the femoral fixed.2. femur was observed pathological observation: HE staining was used to observe the pathology of the femur of rats. .3. femur bone mineral density (BMD) determination: DEXA bone densitometer detection of femur bone density.4. using enzyme linked immunosorbent assay (ELISA) detection of rat serum osteocalcin (BGP), bone morphogenetic protein -2 (BMP-2) concentration level.5. using enzyme linked immunosorbent assay (ELISA) detection of rat serum inflammatory cytokines tumor necrosis factor alpha (TNF- alpha) and interleukins - beta (IL-1 beta) content 6. colorimetric determination of serum calcium (Ca) concentration; phospho molybdic acid determination of serum phosphorus (P) concentration.7. clustering analysis of fecal intestinal microflora by DGGE-PCR technology. Results: 1.HE staining results: normal control group of bone trabecula dense, regular and coarse and uniform; and alcohol model group of rat bone trabecula Loose, irregular, uneven, even bone trabecular fracture; ursolic acid, high dose group compared with the alcohol model group, bone trabecula dense, regular, thick, uniform, no bone trabecular fracture; ursolic acid low dose group improved.2. femur bone density: compared with normal control group, alcohol model group rats The BMD of the femur was significantly reduced and the difference was statistically significant (P0.05). But after 8 weeks of ursolic acid dry prognosis, the BMD of the femur in the high dose group of ursolic acid, except the low dose ursolic acid group, was significantly higher than that in the alcohol model group (P0.05).3. serum BGP, BMP-2 results: the alcohol model group was compared with the normal control group. The content of BGP was significantly reduced and the difference was statistically significant (P0.05). In ursolic acid, the content of BGP increased in the high dose group compared with the alcohol model group, and the difference was statistically significant (P0.05), but there was no statistical difference between the low dose ursolic acid group and the alcohol model group, only the increase in the numerical value. The alcohol model group and the normal control group were compared with the alcohol model group. Compared with the group, the serum BMP-2 content of the rats in the model group was significantly lower, and the difference was statistically significant (P0.05), but the serum BMP-2 content in the high dose ursolic acid group was significantly higher than that in the alcohol model group, and there was a statistically significant difference (P0.05), but the ursolic acid was low and the serum BMP-2 level in the middle dose group changed little, and there was no statistical difference compared with the alcohol model group. .4. serum Ca, P results: the serum Ca, P water in the alcohol model group decreased significantly, compared with the normal control group, the difference was statistically significant (P0.05). Compared with the alcohol model group, the serum Ca and P content of the high dose group increased significantly, and all had statistical difference (P0.05), but the low dose ursolic acid group Ca, P content Although there was an increase in numerical value, there was no statistical difference in the results of.5. serum TNF- A and IL-1 beta: compared with the normal control group, the level of TNF- alpha and IL-1 beta in the alcohol model group was significantly higher, the difference was statistically significant (P0.05), while ursolic acid intervention significantly inhibited the increase of inflammatory factors, compared with the alcohol model group, the level of TNF- A and IL-1 beta. The effect of ursolic acid high dose group was obvious, and the effect of high dose ursolic acid group was obvious, and there was statistical difference (P0.05).6. intestinal microflora DGGE-PCR fingerprint analysis: the intestinal microflora diversity of rats in the alcohol model group was significantly reduced and the similarity with the normal group was lower; and the intestinal flora structure of the high dose ursolic acid group was closer to the normal group. The conclusion: 1. links. The measurement of bone mineral density and pathological observation of femur in rats showed that ursolic acid significantly improved osteoporosis in rats induced by alcohol. The protective mechanism of ursolic acid to alcohol induced osteoporosis could promote bone formation, reduce bone absorption, inhibit bone mineral loss and inflammatory cytokine overlay.2.. .3. ursolic acid can regulate the decrease of intestinal microflora caused by alcohol intake, suggesting that its effect on improving alcoholic osteoporosis may be related to the regulation of intestinal microflora.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R580

【參考文獻】

相關期刊論文 前10條

1 邱奕雁;朱峰;藍濤;陳揚;楊欣建;;IL-6,IL-1β及TNF-α與大鼠骨質(zhì)疏松形成的關系[J];現(xiàn)代生物醫(yī)學進展;2016年18期

2 梁惠;呂銳;傅泳;周治彤;劉穎;周曉彬;王文成;劉曼;馬愛國;;益生菌對大鼠酒精性肝損傷的保護作用及機制研究[J];中國藥理學通報;2016年07期

3 劉雁飛;韓盼;賴永繼;;熊果酸對慢性非細菌性前列腺炎模型大鼠的治療作用[J];醫(yī)藥導報;2015年10期

4 盧新星;范秋靈;徐莉;李琳;徐艷艷;張東成;王力寧;;烏索酸改善高糖誘導系膜細胞損傷的機制[J];中華腎臟病雜志;2015年01期

5 宮笑微;李榮濱;高飛;許雪蓮;朱輝;吳濤;于海躍;宋健;張寒凝;;黑龍江省西部地區(qū)骨質(zhì)疏松流行病學調(diào)查[J];齊齊哈爾醫(yī)學院學報;2014年21期

6 戈娜;梁惠;劉穎;宮安靜;王文成;;海兔素對慢性酒精性肝損傷大鼠肝超微結(jié)構及NO和iNOS的影響[J];中國海洋藥物;2014年03期

7 孫愛平;孫書明;張國俊;王艷華;宋向鳳;;熊果酸減輕LPS誘導的THP-1細胞損傷[J];基礎醫(yī)學與臨床;2014年01期

8 曾光;陳芳;熊新貴;梁清華;陳婷婷;宋煒熙;黃惠勇;;熊果酸對CIA大鼠關節(jié)炎癥及骨質(zhì)破壞的影響[J];湖南中醫(yī)藥大學學報;2013年07期

9 任樹軍;邢國利;葛明富;姜益常;畢旭偉;;補腎健脾益氣法對酒精性骨質(zhì)疏松大鼠小腸中VDR-mRNA表達影響的研究[J];中醫(yī)藥信息;2013年02期

10 ;原發(fā)性骨質(zhì)疏松癥診治指南(2011年)[J];中華骨質(zhì)疏松和骨礦鹽疾病雜志;2011年01期

，

本文編號：2049090