【摘要】：目的有研究表明幽門(mén)螺桿菌感染相關(guān)性胃癌起源于骨髓源性細(xì)胞—骨髓基質(zhì)干細(xì)胞(Bone marrow mesenchymal stem cells , MSCs)。MSCs具有高度自我更新和多向分化的雙重特性。正常環(huán)境中,MSCs很好地保持著靜息和自身增殖分化的平衡。后者的失衡是否和H. pylori相關(guān)的胃癌有關(guān)系未見(jiàn)報(bào)道。我們通過(guò)體外實(shí)驗(yàn)了解H. pylori對(duì)MSCs增殖的影響,來(lái)初步探索MSCs參與H. pylori慢性感染致胃癌發(fā)生發(fā)展的可能機(jī)制。 方法(1)培養(yǎng)純化BALB/C小鼠原代MSCs并鑒定;(2)液體培養(yǎng)H. pylori (TN2菌株);(3)H. pylori培養(yǎng)上清液、超聲粉碎菌體及正常H. pylori液體培養(yǎng)基與MSCs共培養(yǎng);(4)MTT法初步檢測(cè)不同濃度(MOI = H. pylori : MSCs =5 : 1、50 : 1、100 : 1)及不同時(shí)間點(diǎn)(6 h、12 h、24 h)H. pylori對(duì)MSCs增殖的影響。并確定兩者作用的最佳時(shí)間;(5)流式細(xì)胞術(shù)(CFSE法)進(jìn)一步檢測(cè)不同濃度及時(shí)間點(diǎn)H. pylori對(duì)MSCs增殖的影響。 結(jié)果(1)成功分離BALB/C小鼠原代MSCs,其第六代細(xì)胞的純度可達(dá)到95%以上。(2)H. pylori液體培養(yǎng)48小時(shí)后可達(dá)到4×108CFU/ml。(3)MTT法結(jié)果表明共培養(yǎng)體系中H. pylori上清液及超聲粉碎菌體各濃度在6 h和12 h促進(jìn)MSCs增殖(P 0.05),尤其上清液及超聲粉碎菌體6 h最為顯著。(4)流式細(xì)胞術(shù)進(jìn)一步顯示以上各濃度的H. pylori上清液及超聲粉碎菌體組在6 h時(shí)促進(jìn)MSCs增殖(P 0.05)。 結(jié)論一定濃度的H. pylori短時(shí)間可促進(jìn)MSCs的增殖,導(dǎo)致MSCs靜息與增殖分化失衡。
[Abstract]:Objective to show that Helicobacter pylori infection associated gastric cancer originated from bone marrow-derived cells-bone marrow stromal stem cells (BMSCs) with dual characteristics of high self-renewal and multiple differentiation. In normal environments, MSCs maintains a good balance between resting and self-proliferation and differentiation. Whether the latter imbalance is associated with H. pylori related gastric cancer has not been reported. We investigated the effect of H. pylori on the proliferation of MSCs in vitro to explore the possible mechanism of MSCs involved in the development of gastric cancer caused by chronic infection of H. pylori. Methods (1) the primary MSCs of BALB/C mice was cultured and identified, (2) the supernatant of H. pylori (TN2 strain); (3) H. pylori was cultured in liquid culture, and the supernatant was cultured in the medium of ultrasonic crushing and normal H.pylori liquid with MSCs. (4) the effects of different concentrations (MOI = H. pylori: MSCs = 5: 1, 50: 1100: 1) and different time points (6 h, 12 h, 24 h) H. pylori) on the proliferation of MSCs were detected by MTT method. (5) flow cytometry (CFSE) was used to detect the effect of H. pylori on the proliferation of MSCs at different concentrations and time points. Results (1) the primary MSCs, of BALB/C mice was isolated successfully. The purity of the sixth passage cells was more than 95%. (2) H. pylori liquid culture could reach 4 脳 10 8 CFU / ml after 48 hours. (3) the results of MTT method showed that the supernatant of H. pylori and the concentration of supersonic comminuted bacteria in co-culture system were found to be in the range of 4 脳 10 8 CFU / ml. The proliferation of MSCs was promoted at 6 h and 12 h (P 0.05). (4) flow cytometry showed that the supernatant of H. pylori supernatant and the supersonic crushing group promoted the proliferation of MSCs at 6 h (P 0.05). Conclusion A certain concentration of H. pylori can promote the proliferation of MSCs for a short time and lead to the imbalance between resting and proliferation and differentiation of MSCs.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R329

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