本文選題：恒古骨傷愈合劑 + 骨質(zhì)疏松癥��； 參考：《貴州醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探究恒古骨傷愈合劑調(diào)控大鼠BMSCs成脂肪細(xì)胞分化的能力及其對(duì)防治骨質(zhì)疏松癥的作用機(jī)制。方法:以大鼠BMSCs為研究對(duì)象,采用全骨髓貼壁培養(yǎng)法,從幼年SD大鼠股骨和脛骨骨髓中分離培養(yǎng)BMSCs;免疫組織細(xì)胞化學(xué)染色檢測BMSCs細(xì)胞純度;恒古骨傷愈合劑分別稀釋500倍、1000倍、2000倍、4000倍,相對(duì)應(yīng)分5組(control組、2×10-3組、1×10-3組、0.5×10-3組、0.25×10-3組),其中control組加入溶媒;MTT法檢測各濃度恒古骨傷愈合劑對(duì)BMSCs細(xì)胞毒性;BMSCs在不同濃度恒古骨傷愈合劑干預(yù)下分別向脂肪細(xì)胞方向誘導(dǎo)分化;在恒古骨傷愈合劑干預(yù)后的第21天,Oil Red O染色后顯微鏡下觀察及酶標(biāo)儀定量檢測成脂肪細(xì)胞分化情況;在恒古骨傷愈合劑干預(yù)后的第7天、14天、21天,Realtime-PCR定量檢測成脂肪細(xì)胞分化特異性轉(zhuǎn)錄因子過氧化物酶體增值物激活受體(PPAR)γ、CCAAT/增強(qiáng)子結(jié)合蛋白(C/EBP)α的mRNA相對(duì)表達(dá)量。結(jié)果:?采用細(xì)胞貼壁生長法獲得大鼠(乳鼠)BMSCs;?免疫組織細(xì)胞化學(xué)染色后熒光倒置顯微鏡下觀察發(fā)現(xiàn)BMSCs特異性表面抗原CD44陽性,陽性率為95.47%,而CD19陰性,提示培養(yǎng)的細(xì)胞為骨髓間充質(zhì)干細(xì)胞且純度高;?MTT法檢測結(jié)果示:在恒古骨傷愈合劑干預(yù)BMSCs后的0.5天、1天、4天、21天,各組與control組比較,差異無統(tǒng)計(jì)學(xué)意義(P0.05)。提示各濃度恒古骨傷愈合劑不論是早期還是晚期,對(duì)BMSCs的增殖及細(xì)胞活性均無毒性作用;?在恒古骨傷愈合劑干預(yù)BMSCs誘導(dǎo)成脂肪細(xì)胞分化的第21天,Oil Red O染色后顯微鏡下觀察發(fā)現(xiàn)各濃度藥物均對(duì)BMSCs成脂肪細(xì)胞分化有抑制作用。成濃度依賴性,且濃度越高抑制作用越強(qiáng)。酶標(biāo)儀定量檢測OD值分別為(1.104±0.414、0.785±0.085、0.836±0.138、1.008±0.369、1.000±0.291),2×10-3組、1×10-3組與control組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),與染色觀察結(jié)果一致;(5)恒古骨傷愈合劑能抑制脂肪細(xì)胞生成,下調(diào)脂肪細(xì)胞特異性轉(zhuǎn)錄因子的表達(dá)。在恒古骨傷愈合劑干預(yù)的第7天、14天、21天,Realtime-PCR定量檢測結(jié)果提示PPARγ、C/EBPα基因表達(dá)量與control組比較下調(diào)且差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:恒古骨傷愈合劑可通過下調(diào)PPARγ、C/EBPα的表達(dá)而抑制大鼠BMSCs成脂肪細(xì)胞分化。抑制BMSCs成脂肪細(xì)胞分化是恒古骨傷愈合劑防治骨質(zhì)疏松癥的另一作用途徑。
[Abstract]:Aim: to investigate the ability of Henggu bone wound healing agent to regulate the differentiation of rat BMSCs adipoblasts and its mechanism of prevention and treatment of osteoporosis.Methods: BMSCs were isolated from femur and tibia bone marrow of young SD rats by whole bone marrow adherent culture, and the purity of BMSCs cells was detected by immunohistochemical staining.Henggu bone wound healing agent diluted 500 times, 1000 times, 2000 times and 4000 times respectively,The differentiation of adipocytes was observed under microscope on the 21st day after the intervention of Henggu bone wound healing agent with oil Red O, and the differentiation of adipocytes was detected quantitatively by enzyme labeling instrument.It turns out to be:?The method of cell adherent growth was used to obtain BMSCs?The positive rate of BMSCs specific surface antigen (CD44) was 95.47%, but CD19 was negative.The results showed that the cultured cells were bone marrow mesenchymal stem cells and the purity was high. The results showed that there was no significant difference between control group and Heng Gu bone wound Healing Agent 0. 5 day, 0. 5 days, 4 days and 21 days, compared with control group, the difference was not statistically significant (P 0. 05).The results suggest that there is no toxic effect on proliferation and cell activity of BMSCs in early or late stage.Oil Red O staining of adipocytes induced by BMSCs on the 21st day after treatment with Henggu bone wound healing agent was observed under microscope. It was found that every concentration of drugs could inhibit the differentiation of BMSCs adipocytes.In a concentration-dependent manner, the higher the concentration, the stronger the inhibitory effect.The quantitative analysis of real time-PCR showed that the expression of PPAR 緯 C / EBP 偽 gene was down-regulated compared with that of control group, and the difference was statistically significant (P 0.05).Conclusion: Henggu bone healing agent can inhibit the differentiation of rat BMSCs adipoblasts by down-regulating the expression of PPAR 緯 -C / EBP 偽.Inhibiting the differentiation of BMSCs adipoblasts is another way to prevent and cure osteoporosis with Henggu bone injury healing agent.
【學(xué)位授予單位】：貴州醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R580

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳光華;黃貴芝;林顥;吳浩俊;陳航;;骨髓間充質(zhì)干細(xì)胞移植對(duì)去卵巢骨質(zhì)疏松大鼠骨密度的影響[J];中國組織工程研究;2017年01期

2 雷鳴;劉春穎;劉曉峰;孫琳;鄭文奎;;人羊膜間充質(zhì)干細(xì)胞移植治療去卵巢骨質(zhì)疏松[J];中國組織工程研究;2017年01期

3 孫巖;陳冉;王小琦;張洋;蔣罡;胡敏;趙宏斌;李文輝;;恒古骨傷愈合劑對(duì)去勢(shì)雌性大鼠骨質(zhì)疏松模型的影響[J];中國骨質(zhì)疏松雜志;2016年12期

4 薛小鑾;陳永法;;芪骨膠囊與仙靈骨葆膠囊治療原發(fā)性骨質(zhì)疏松癥的Meta分析[J];中國藥物經(jīng)濟(jì)學(xué);2016年11期

5 王冬生;韓婧;康文博;趙建寧;;植物雌激素防治骨質(zhì)疏松作用的機(jī)制進(jìn)展[J];中國骨質(zhì)疏松雜志;2016年05期

6 楊鋒;潘樂;馬秋濤;張濤;楊利學(xué);李彥民;;右歸丸對(duì)骨髓間充質(zhì)干細(xì)胞成脂分化相關(guān)基因甲基化的影響[J];中國老年學(xué)雜志;2015年24期

7 莫林海;莊寧;劉成龍;羅瑜;李松建;楊少華;趙海軍;劉建休;黃世嘉;;外源性NGF對(duì)轉(zhuǎn)基因CGRP干細(xì)胞移植治療骨質(zhì)疏松大鼠的影響[J];中國骨質(zhì)疏松雜志;2015年12期

8 王劍;王鋼;陳天宇;格日勒?qǐng)D;;異補(bǔ)骨脂素對(duì)小鼠骨髓基質(zhì)干細(xì)胞成骨和成脂分化影響及其機(jī)制的相關(guān)研究[J];中華創(chuàng)傷骨科雜志;2015年12期

9 徐錚;顧昊;李冠武;常時(shí)新;周蕾;;唑來膦酸抑制骨質(zhì)疏松骨髓脂肪增多[J];中國骨質(zhì)疏松雜志;2015年08期

10 王素彬;趙宏斌;胡敏;魏蔚;錢傳云;蔣罡;張洋;趙林;郭磊;;恒古骨傷愈合劑對(duì)骨質(zhì)疏松性骨折兔骨密度及DKK-1蛋白的影響[J];中國中西醫(yī)結(jié)合雜志;2015年08期

相關(guān)博士學(xué)位論文 前1條

1 趙王林;淫羊藿苷對(duì)骨質(zhì)疏松大鼠MSCs定向分化干預(yù)的實(shí)驗(yàn)研究[D];廣州中醫(yī)藥大學(xué);2011年

相關(guān)碩士學(xué)位論文 前3條

1 張小強(qiáng);杜仲提取物5-羥甲基-2-糠醛誘導(dǎo)細(xì)胞成脂分化的信號(hào)途徑研究[D];南京中醫(yī)藥大學(xué);2016年

2 郭磊;恒古骨傷愈合劑對(duì)骨質(zhì)疏松兔力學(xué)及微結(jié)構(gòu)影響的基礎(chǔ)研究[D];昆明醫(yī)科大學(xué);2014年

3 陳晚嬌;蛇床子素對(duì)大鼠骨髓間充質(zhì)干細(xì)胞增殖和脂向分化的影響[D];暨南大學(xué);2007年

，

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