本文選題：間充質(zhì)干細(xì)胞　切入點(diǎn)：低劑量照射　出處：《蘇州大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：研究低劑量X線照射對離體培養(yǎng)的骨髓間充質(zhì)干細(xì)胞增殖和分化的影響，探討低劑量照射促進(jìn)大鼠骨折后骨痂礦化的細(xì)胞機(jī)制。 方法：1.由大鼠骨髓中提取間充質(zhì)干細(xì)胞（BMSCs），分別以0Gy、25mGy、50mGy、75mGy、100mGy、1000mGy不同劑量的X線照射，以CCK-8法檢測照射6h、12h、24h、48h、72h后細(xì)胞的增殖情況，以流式細(xì)胞技術(shù)檢測照射后對細(xì)胞周期的影響。2.如前述劑量照射后，誘導(dǎo)BMSCs向成骨細(xì)胞方向分化，評估分化的成骨細(xì)胞的成骨潛能。以Vonkossa染色法計(jì)數(shù)礦化結(jié)節(jié)的形成數(shù)目并評估其成骨能力。以RT-PCR法檢測成骨相關(guān)基因OPG、COL-1、BGP的表達(dá)情況。 結(jié)果：1.CCK-8法檢測結(jié)果表明75mGy低劑量照射后，可明顯刺激BMSCs的增殖，與假照射組相比有統(tǒng)計(jì)學(xué)意義（P＜0.05）。而1000mGy高劑量照射則會抑制BMSCs增殖（P＜0.05）。流式細(xì)胞技術(shù)檢測表明，，低劑量X線照射可以促進(jìn)BMSCs由G1期進(jìn)入S期，與假照射組相比具有統(tǒng)計(jì)學(xué)意義（P＜0.05）。2.向成骨細(xì)胞誘導(dǎo)分化后，低劑量照射組礦化結(jié)節(jié)數(shù)目較假照射組明顯增多，以75mGy與100mGy組最為明顯（P＜0.05）。RT-PCR結(jié)果顯示OPG、COL-1、BGP經(jīng)低劑量X線照射后表達(dá)量均有明顯增高，以以75mGy與100mGy組最為明顯（P＜0.05），而1000mGy高劑量照射則會抑制相關(guān)成骨基因的表達(dá)。 結(jié)論：1.低劑量X線照射可有效促進(jìn)BMSCs的增殖，而高劑量照射則有抑制作用。2.低劑量X線照射可增強(qiáng)BMSCs的分化能力，照射后BMSCs成骨潛能明顯增強(qiáng)，而高劑量照射起到相反的作用。
[Abstract]:Aim: to study the effects of low dose X-ray irradiation on the proliferation and differentiation of bone marrow mesenchymal stem cells (MSCs) cultured in vitro and to explore the cellular mechanism of bone callus mineralization induced by low dose irradiation. Methods 1. Bone marrow mesenchymal stem cells (BMSCs) were extracted from rat bone marrow and irradiated with different doses of 0 Gy 25 mGy 50 mGy 75 mGy 100 mGy respectively. The proliferation of cells was detected by CCK-8 method after irradiation for 6 h 12 h or 24 h or 48 h for 72 h. Flow cytometry was used to detect the effect of irradiation on cell cycle. (2) BMSCs was induced to differentiate into osteoblasts, The osteogenic potential of differentiated osteoblasts was evaluated. The number of mineralized nodules was counted and the osteogenic ability was evaluated by Vonkossa staining. The expression of osteoblastic associated gene OPGG-COL-1 was detected by RT-PCR method. Results 1. The results of CCK-8 assay showed that 75 mGy could significantly stimulate the proliferation of BMSCs after low dose irradiation, which was significantly higher than that of sham irradiation group (P < 0.05), while high dose irradiation of 1000mGy could inhibit the proliferation of BMSCs (P < 0.05). Flow cytometry showed that the proliferation of BMSCs was inhibited by high dose irradiation (P < 0.05). Low dose X-ray irradiation could promote BMSCs from G1 phase to S phase. Compared with false irradiation group, the number of mineralized nodules in low dose irradiation group was significantly higher than that in false irradiation group (P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05). The results of RT-PCR showed that the expression of BGP in 75mGy and 100mGy groups was significantly increased after low dose X-ray irradiation, especially in 75mGy and 100mGy groups (P < 0.05), while the expression of osteogenic genes was inhibited by high dose radiation of 1000mGy. Conclusion low dose X ray irradiation can effectively promote the proliferation of BMSCs, while high dose X ray irradiation can inhibit the proliferation of BMSCs. Low dose X ray irradiation can enhance the differentiation ability of BMSCs, and the osteogenic potential of BMSCs is obviously enhanced after irradiation. High doses of radiation have the opposite effect.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329

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