本文關(guān)鍵詞：運動對骨質(zhì)疏松小鼠骨長鏈非編碼RNA表達的影響　出處：《上海體育學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 骨質(zhì)疏松 運動 骨密度 lncRNA 骨生物力學(xué) 骨形態(tài)計量學(xué)

【摘要】：研究目的:骨質(zhì)疏松是一種以全身性骨量減少以及骨組織微觀結(jié)構(gòu)退化為特征的疾病,會導(dǎo)致骨強度降低,脆性增加,骨折風(fēng)險增加。采用運動方法用于骨質(zhì)疏松的預(yù)防和治療可以有效的減少因藥物治療帶來的家庭經(jīng)濟負擔(dān)和社會醫(yī)療保障系統(tǒng)的負擔(dān)及其毒副作用。運動可以通過機械應(yīng)力刺激、細胞因子、骨代謝信號通路等對骨質(zhì)疏松的防治發(fā)揮關(guān)鍵作用,其作用機制是當(dāng)下一大研究熱點。最新研究表明非編碼RNA(long noncoding RNA,lncRNA)在骨代謝中發(fā)揮重要的調(diào)控作用。本研究擬通過建立小鼠骨質(zhì)疏松模型,并進行中等強度的運動干預(yù),探究運動對骨質(zhì)疏松小鼠骨lncRNA表達的影響,為進一步探究運動是否會通過影響骨lncRNA起到防治骨質(zhì)疏松的作用提供理論依據(jù)。研究方法:8周齡雌性C57BL/6小鼠44只,隨機分為假手術(shù)組(SHAM),假手術(shù)+運動組(SHAM+EX),去卵巢組(OVX),去卵巢+運動組(OVX+EX)。去卵巢小鼠麻醉后于背部脊柱旁開1cm,身長下1/3處切口去除卵巢,假手術(shù)小鼠于相同部位去除同等大小的脂肪。手術(shù)后4周,SHAM+EX組和OVX+EX組進行為期9周的跑臺訓(xùn)練,每周5天,周六周日休息,坡度25°。第一周為運動預(yù)適應(yīng),速度6M/min,時間30min;后八周起始速度為8M/min,每周增加1M/min;時間為5min熱身運動(速度:6M/min)和55min的正式訓(xùn)練。SHAM組和OVX組不進行任何干預(yù),常規(guī)飼養(yǎng)。干預(yù)結(jié)束后取血尿以及左右股骨脛骨,左側(cè)股骨用于檢測骨密度、mcro CT以及骨生物力學(xué)指標(biāo),右側(cè)股骨檢測骨形態(tài)計量學(xué)指標(biāo),每組隨機選取3只小鼠的全部股骨脛骨,提取總RNA用于lncRNA芯片檢測,同時對各組差異性表達在4倍以上的lncRNA進行篩選用于GO Pathway分析。所得數(shù)據(jù)結(jié)果用SPSS 20.0軟件進行統(tǒng)計分析。研究結(jié)果:(1)去卵巢可以使小鼠體重增加明顯,運動可以減少去卵巢小鼠的體重。(2)運動對骨密度的影響:運動可以顯著提高假手術(shù)與骨質(zhì)疏松小鼠的BMD,與SHAM組相比,SHAM+EX組BMD顯著提高(P0.05),OVX組顯著降低(P0.01);與OVX組相比,OVX+EX組BMD顯著提高(P0.01)。(3)運動對骨質(zhì)疏松小鼠骨形態(tài)計量學(xué)的影響:SHAM組的MAR低于SHAM+EX組但沒有顯著差異,顯著高于OVX組(P0.05),OVX組MAR顯著低于OVX+EX組(P0.05),運動可以提高假手術(shù)和骨質(zhì)疏松的小鼠的BFR/BV,但沒有顯著性差異。與SHAM組相比,SHAM+EX組小鼠的OB.S/BS顯著提高(P0.05),OVX組顯著降低(P0.01);與OVX組相比,OVX+EX組OB.S/BS顯著提高(P0.05)。與SHAM組相比,OVX組OC.S/BS顯著增加(P0.01);與OVX組相比,OVX+EX組OC.S/BS顯著降低(P0.01)。(4)microCT檢測:運動可以明顯提高假手術(shù)與骨質(zhì)疏松小鼠的BV/TV、Tb.Th、Tb.N降低Tb.Sp,但結(jié)果沒有顯著性差異。(5)運動對骨質(zhì)疏松小鼠骨生物力學(xué)的影響:與SHAM組相比,SHAM+EX組最大載荷顯著增加(P0.05),OVX組顯著降低(P0.05);與OVX組相比,OVX+EX組最大載荷顯著增加(P0.05)。與SHAM組相比,SHAM+EX組彎曲強度提高但無顯著差異,OVX組顯著降低(P0.05),與OVX相比OVX+EX組顯著降低(P0.05)。與SHAM相比,SHAM+EX組彈性模量提高但無顯著差異,OVX+EX組彈性模量較OVX組提高,但無顯著差異。(6)運動對骨質(zhì)疏松小鼠骨lncRNAs的影響:篩選出差異性表達在2倍和4倍以上,且P0.05的lncRNAs。與SHAM組相比,SHAM+EX組差異表達在2倍以上的有64條表達上調(diào),161條表達下調(diào),差異在4倍以上表達上調(diào)的有3條,表達下調(diào)的有3條。與SHAM組相比,OVX組差異性表達2倍以上的有437條表達上調(diào),其中Gm30783、LOC102639568等25條差異性在4倍以上;有231條表達下調(diào),其中Gm33272、Gm12678、Gm10248、LOC105243625等17條差異性在4倍以上。與OVX組相比,OVX+EX組差異性表達在2倍以上的有181條表達下調(diào),其中l(wèi)nc 105247265等3條lncRNA的差異性在4倍以上;56條表達上調(diào),其中Gm31126、LOC105243692、Gm11651等8條的差異性在4倍以上。(7)通過KEGG pathway分析發(fā)現(xiàn),Melanoma信號通路、mTOR信號通路、Toxoplasmosis信號通路、NF-κB信號通路、PI3K-Akt signaling pathway等信號通路可能被lncRNA調(diào)控。Regulation of cell cycle phase transition、vocalization behavior、detection of biotic stimulus等生物過程,TOR complex、synapse part等細胞組分以VEGF-A-activated receptor activity、VEGF-B-activated receptor activity等分子可能被lncRNA所調(diào)控。研究結(jié)論:中等強度的跑臺運動可以顯著提高骨質(zhì)松小鼠的骨密度,提高其骨生成速率和骨強度;同時可以對小鼠骨lncRNA產(chǎn)生一定的影響,造成其表達的升高或降低,但其機制有待進一步研究。
[Abstract]:Objective: osteoporosis is a decrease and degradation of the microstructure of bone tissue disease characterized by systemic bone, will lead to reduced bone strength, brittleness increases, increase the risk of fracture. The exercise method for the prevention and treatment of osteoporosis due to drug treatment can effectively reduce the financial burden of the family and social medical security system the burden and side effects. They can move through mechanical stress, cell factor, prevention and treatment of bone metabolic pathways on osteoporosis play a key role in the mechanism is a hot research topic. The latest research shows that non encoding RNA (long noncoding RNA, lncRNA) play an important role in the regulation of bone metabolism in the present study was to establish a mouse model for osteoporosis, exercise intervention and moderate intensity, explore the impact of exercise on bone osteoporosis lncRNA expression in mice, as in Further study whether exercise would provide a theoretical basis for the effect on the bone lncRNA contributes to the prevention and treatment of osteoporosis. Methods: 8 week old female C57BL/6 44 mice were randomly divided into sham operation group (SHAM), sham operation + exercise group (SHAM+EX), ovariectomized group (OVX), OVX + exercise group (OVX+EX) in ovariectomized mice. After anesthesia 1cm on the back of paravertebral, length 1/3 incision at the removal of the ovaries, sham operated mice in the same part of the same size to remove fat. 4 weeks after surgery, SHAM+EX group and OVX+EX group for 9 weeks of treadmill training, 5 days a week, Saturday Sunday rest, slope 25 degrees. The first week of exercise preconditioning, speed 6M/min, time 30min; eight weeks after the initial velocity is 8M/min, the weekly increase 1M/min; time is 5min warm-up (speed: 6M/min) and 55min.SHAM formal training group and OVX group without any intervention, regular feeding. After intervention for hematuria and The left and right femur tibia, left femur for the detection of bone mineral density and bone biomechanics, mcro CT index, index of right femur bone histomorphometry detection, each group of 3 mice were randomly selected and all femoral and tibial total RNA extraction, lncRNA chip for detection, at the same time to each expression of more than 4 times in lncRNA were screened for GO Pathway analysis the data obtained by SPSS 20. The results were analyzed. Results: (1) ovariectomy can make the body weight of mice increased significantly, exercise can reduce the weight of ovariectomized mice. (2) the effects of exercise on bone mineral density: exercise can significantly improve the sham operation and osteoporosis in mice BMD, compared with the SHAM group SHAM+EX group BMD significantly increased (P0.05), OVX group significantly decreased (P0.01); compared with OVX group, OVX+EX group BMD significantly increased (P0.01). (3) effects of exercise on bone histomorphometry in osteoporosis mice: group SHAM MAR Lower than the SHAM+EX group but no significant differences, significantly higher than that of group OVX (P0.05), OVX MAR group was significantly lower than that of OVX+EX group (P0.05), sham operation and exercise can improve the osteoporosis in mice BFR/BV, but no significant difference. Compared with the SHAM group, SHAM+EX group of mice OB.S/BS significantly increased (P0.05), OVX group significantly decreased (P0.01); compared with OVX group, OVX+EX group OB.S/BS significantly increased (P0.05). Compared with SHAM group, OVX group, OC.S/BS increased significantly (P0.01); compared with OVX group, OVX+EX group, OC.S/BS decreased significantly (P0.01). (4) microCT detection: exercise can significantly improve the sham operation and osteoporosis in mice BV/TV, Tb.Th, Tb.N decreased Tb.Sp, but the results had no significant difference. (5) the effects of exercise on bone biomechanics in osteoporosis mice: compared with SHAM group, SHAM+EX group significantly increased the maximum load (P0.05), OVX group significantly decreased (P0.05); compared with OVX group, OVX+EX group, the maximum load significantly Increased (P0.05). Compared with the SHAM group, but no significant difference between the SHAM+EX group to improve the bending strength of OVX group decreased significantly (P0.05), compared with OVX OVX+EX group decreased significantly (P0.05). Compared with SHAM, but there was no significant difference between the SHAM+EX group to improve the elastic modulus, elastic modulus of OVX+EX group than in OVX group increased, but no significant difference. (6) the effects of exercise on bone lncRNAs osteoporosis mice: screening travelheterosexual expression in 2 times and 4 times, P0.05 and lncRNAs. compared with group SHAM, group SHAM+EX differentially expressed in more than 2 times the 64 up-regulated genes and 161 downregulated and up-regulated the expression of differences in more than 4 times of the 3, down regulated 3. Compared with SHAM group, OVX group, differential expression of 2 times more than the 437 up-regulated genes including Gm30783, LOC102639568, etc. 25 differences in more than 4 times; 231 expression, including Gm33272, Gm12678, Gm10248, LOC105243625 and other 17 difference specific In more than 4 times. Compared with the OVX group, OVX+EX group differentially expressed in more than 2 times of the 181 expression, and the difference of LNC 105247265 and 3 lncRNA in more than 4 times; 56 up-regulated genes including Gm31126, LOC105243692, Gm11651, etc. 8 of the differences in the (more than 4 times. 7) by KEGG pathway analysis showed that the Melanoma signaling pathway, mTOR signaling pathway, Toxoplasmosis signaling pathway, B signaling pathway NF-, PI3K-Akt signaling pathway signaling pathway may be regulated by lncRNA.Regulation of cell cycle phase transition, vocalization behavior, detection of biotic stimulus TOR complex synapse, biological process, and cellular component in part VEGF-A-activated receptor activity, VEGF-B-activated receptor activity and other molecules may be regulated lncRNA. Conclusion: moderate intensity treadmill exercise can significantly improve the osteoporosis mice bone Density can increase the rate of bone formation and bone strength, meanwhile, it can produce certain effects on the bone lncRNA in mice, resulting in the increase or decrease of its expression, but its mechanism needs further study.

【學(xué)位授予單位】：上海體育學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R580

【參考文獻】

相關(guān)期刊論文 前10條

1 李世昌;李靈杰;陳祥和;孫朋;趙常紅;徐帥;方幸;;運動對生長期小鼠骨代謝及PDGF介導(dǎo)的ERK信號通路的影響[J];沈陽體育學(xué)院學(xué)報;2016年06期

2 張玲莉;元宇;鄒軍;;機械載荷對骨組織骨形態(tài)發(fā)生蛋白信號通路的影響[J];中國細胞生物學(xué)學(xué)報;2016年08期

3 陳熙;郭健民;元宇;張玲莉;吳雋旎;孫忠廣;陳炳霖;鄒軍;;不同牽張應(yīng)力對成骨細胞MC3T3-E1分化及Wnt信號轉(zhuǎn)導(dǎo)通路的影響研究[J];中國骨質(zhì)疏松雜志;2016年01期

4 常波;高海寧;佟瑤;李濤;王卉瑩;武燕;賀佳;曲廣才;;運動和藥物聯(lián)合干預(yù)對老年2型糖尿病大鼠骨密度和骨生物力學(xué)的影響[J];沈陽體育學(xué)院學(xué)報;2015年06期

5 鄭陸;陳曉紅;李荀;王智強;王文榮;;中等強度運動ERβ和RANKL/OPG變化及對骨量的調(diào)控機制[J];西安體育學(xué)院學(xué)報;2015年05期

6 馬濤;李世昌;;上、下坡跑臺運動對去卵巢小鼠破骨細胞分化NF-κB信號通路的影響[J];中國運動醫(yī)學(xué)雜志;2015年05期

7 高艷;程晨;李靜;肖偉凡;潘秋輝;;LncRNA AK051397在BMP-2誘導(dǎo)的C2C12成骨分化中的作用[J];同濟大學(xué)學(xué)報(醫(yī)學(xué)版);2014年04期

8 許澳;沈晶;卜淑敏;謝金珍;韓天雨;;跑臺運動訓(xùn)練和停訓(xùn)對去卵巢大鼠體成分和骨密度的影響[J];現(xiàn)代生物醫(yī)學(xué)進展;2014年03期

9 賈曉凡;孫明曉;嚴(yán)翊;李博文;仲偉娟;陳俊飛;張一民;王正珍;王璐;謝敏豪;;有氧運動對超重青少年骨密度的影響[J];中華骨質(zhì)疏松和骨礦鹽疾病雜志;2012年03期

10 李世昌;季瀏;劉體偉;馬濤;陳祥和;;不同方式運動對去卵巢大鼠骨密度、骨生物力學(xué)及代謝指標(biāo)的影響[J];體育學(xué)刊;2012年02期

相關(guān)碩士學(xué)位論文 前1條

1 王錦玉;小鼠成骨細胞特異性長鏈非編碼RNA影響其功能的研究[D];中南大學(xué);2014年

，

本文編號：1374559