幽门螺杆菌对骨髓基质干细胞增殖的影响
发布时间：2019-02-17 08:46
【摘要】：目的有研究表明幽门螺杆菌感染相关性胃癌起源于骨髓源性细胞—骨髓基质干细胞(Bone marrow mesenchymal stem cells , MSCs)。MSCs具有高度自我更新和多向分化的双重特性。正常环境中,MSCs很好地保持着静息和自身增殖分化的平衡。后者的失衡是否和H. pylori相关的胃癌有关系未见报道。我们通过体外实验了解H. pylori对MSCs增殖的影响,来初步探索MSCs参与H. pylori慢性感染致胃癌发生发展的可能机制。 方法(1)培养纯化BALB/C小鼠原代MSCs并鉴定;(2)液体培养H. pylori (TN2菌株);(3)H. pylori培养上清液、超声粉碎菌体及正常H. pylori液体培养基与MSCs共培养;(4)MTT法初步检测不同浓度(MOI = H. pylori : MSCs =5 : 1、50 : 1、100 : 1)及不同时间点(6 h、12 h、24 h)H. pylori对MSCs增殖的影响。并确定两者作用的最佳时间;(5)流式细胞术(CFSE法)进一步检测不同浓度及时间点H. pylori对MSCs增殖的影响。 结果(1)成功分离BALB/C小鼠原代MSCs,其第六代细胞的纯度可达到95%以上。(2)H. pylori液体培养48小时后可达到4×108CFU/ml。(3)MTT法结果表明共培养体系中H. pylori上清液及超声粉碎菌体各浓度在6 h和12 h促进MSCs增殖(P 0.05),尤其上清液及超声粉碎菌体6 h最为显著。(4)流式细胞术进一步显示以上各浓度的H. pylori上清液及超声粉碎菌体组在6 h时促进MSCs增殖(P 0.05)。 结论一定浓度的H. pylori短时间可促进MSCs的增殖,导致MSCs静息与增殖分化失衡。
[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.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
本文编号：2424982
[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.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
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