HMGB1对体外人肺动脉平滑肌细胞增殖、迁移和凋亡的影响
发布时间：2019-03-01 14:06
【摘要】：目的：观察高迁移率族蛋白B1(HMGB1)对体外人肺动脉平滑肌细胞(HPASMC)增殖、迁移和凋亡的影响。 方法：体外培养人肺动脉平滑肌细胞(HPASMC)至5～8代,然后开始实验。配制含不同浓度HMGB1 (Ong/ml、1 ng/ml、10 ng/ml、100 ng/ml和1000 ng/ml)的培养液,体外分别培养24小时和48小时后,用四甲基偶氮唑蓝(MTT)法检测不同浓度的HMGB1对HPASMC细胞生长增殖率的影响。利用Boyden小室法检测穿膜细胞数,观察含不同浓度HMGB1的无血清培养基对HPASMC体外培养20小时的趋化作用。用TUNEL法检测不同浓度HMGB1对HPASMC作用48小时后细胞凋亡情况的影响。 结果：(1)MTT法检测显示HMGB1对HPASMC细胞作用24小时或48小时后,与阴性对照组相比,实验组细胞增殖率明显升高,随着HMGB1浓度增加,细胞增殖率升高(P0.01),并在HMGB1浓度为100ng/ml时达到平台,且经HMGB1处理48小时后细胞增殖率高于24小时对应浓度组(P0.05)。(2)Boyden小室法趋化实验显示随着HMGB1浓度升高,穿膜细胞数增加(P0.01),且HMGB1浓度为100 ng/ml时趋化作用达到峰值,HMGB1浓度为1μg/ml时趋化作用略减弱。(3) TUNEL法检测细胞凋亡结果显示,不同浓度HMGB1 (Ong/mL\1 ng/ml、10 ng/ml、100 ng/ml和1000 ng/ml)干预48小时后,对照组和各实验组的细胞凋亡率分别为8.56%,8.22%,7.61%,7.50%和7.11%(P0.05)。 结论：1.HMGB1具有促进HPASMC进行细胞增殖和迁移的作用,且该作用对HMGB1具有浓度依赖性,在HMGB1浓度为100ng/ml时达到峰值。 2. HMGB1对HPASMC的细胞凋亡无明显影响。 3. HMGB1通过诱导HPASMC增殖和迁移来促进肺血管重构。
[Abstract]:Aim: to observe the effects of high mobility group B1 (HMGB1) on proliferation, migration and apoptosis of human pulmonary artery smooth muscle cells (HPASMC) in vitro. Methods: human pulmonary artery smooth muscle cells (HPASMC) were cultured in vitro for 5 ~ 8 passages, and then the experiment was started. The culture medium containing different concentrations of HMGB1 (Ong/ml,1 ng/ml,10 ng/ml,100 ng/ml and 1000 ng/ml) was prepared and cultured in vitro for 24 hours and 48 hours, respectively. The effects of different concentrations of HMGB1 on the growth and proliferation of HPASMC cells were detected by tetramethylazo blue (MTT) assay. Boyden chamber method was used to detect the number of transmembrane cells. The chemotactic effect of serum-free medium containing different concentrations of HMGB1 on HPASMC in vitro for 20 hours was observed. TUNEL assay was used to detect the effect of different concentrations of HMGB1 on apoptosis after 48 hours of HPASMC treatment. Results: (1) MTT assay showed that the proliferation rate of HPASMC cells treated with HMGB1 for 24 hours or 48 hours was significantly higher than that of the negative control group. With the increase of HMGB1 concentration, the cell proliferation rate increased (P0.01). When the concentration of HMGB1 was 100ng/ml, the cell proliferation rate was higher than that of the corresponding concentration group (P0.05). (2) Boyden chamber chemotaxis test showed that with the increase of HMGB1 concentration, the cell proliferation rate after 48 hours of HMGB1 treatment was higher than that of the corresponding concentration group (P0.05). (2). The number of transmembrane cells was increased (P0.01), and the chemotaxis reached the peak when HMGB1 concentration was 100 ng/ml, and the chemotaxis was slightly decreased when HMGB1 concentration was 1 渭 g / ml. (3) the results of TUNEL assay showed that the chemotaxis reached the peak value. After 48 hours of intervention with different concentrations of HMGB1 (Ong/mL\ 1 ng/ml,10 ng/ml,100 ng/ml and 1000 ng/ml), the apoptosis rates of the control group and each experimental group were 8.56%, 8.22%, 7.61%, respectively. 7.50% and 7.11% (P0.05). Conclusion: 1.HMGB1 can promote the proliferation and migration of HPASMC cells, and it has a concentration-dependent effect on HMGB1 and reaches its peak when the concentration of HMGB1 is 100ng/ml. 2. HMGB1 had no effect on the apoptosis of HPASMC. 3. HMGB1 promotes pulmonary vascular remodeling by inducing HPASMC proliferation and migration.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
本文编号：2432494
[Abstract]:Aim: to observe the effects of high mobility group B1 (HMGB1) on proliferation, migration and apoptosis of human pulmonary artery smooth muscle cells (HPASMC) in vitro. Methods: human pulmonary artery smooth muscle cells (HPASMC) were cultured in vitro for 5 ~ 8 passages, and then the experiment was started. The culture medium containing different concentrations of HMGB1 (Ong/ml,1 ng/ml,10 ng/ml,100 ng/ml and 1000 ng/ml) was prepared and cultured in vitro for 24 hours and 48 hours, respectively. The effects of different concentrations of HMGB1 on the growth and proliferation of HPASMC cells were detected by tetramethylazo blue (MTT) assay. Boyden chamber method was used to detect the number of transmembrane cells. The chemotactic effect of serum-free medium containing different concentrations of HMGB1 on HPASMC in vitro for 20 hours was observed. TUNEL assay was used to detect the effect of different concentrations of HMGB1 on apoptosis after 48 hours of HPASMC treatment. Results: (1) MTT assay showed that the proliferation rate of HPASMC cells treated with HMGB1 for 24 hours or 48 hours was significantly higher than that of the negative control group. With the increase of HMGB1 concentration, the cell proliferation rate increased (P0.01). When the concentration of HMGB1 was 100ng/ml, the cell proliferation rate was higher than that of the corresponding concentration group (P0.05). (2) Boyden chamber chemotaxis test showed that with the increase of HMGB1 concentration, the cell proliferation rate after 48 hours of HMGB1 treatment was higher than that of the corresponding concentration group (P0.05). (2). The number of transmembrane cells was increased (P0.01), and the chemotaxis reached the peak when HMGB1 concentration was 100 ng/ml, and the chemotaxis was slightly decreased when HMGB1 concentration was 1 渭 g / ml. (3) the results of TUNEL assay showed that the chemotaxis reached the peak value. After 48 hours of intervention with different concentrations of HMGB1 (Ong/mL\ 1 ng/ml,10 ng/ml,100 ng/ml and 1000 ng/ml), the apoptosis rates of the control group and each experimental group were 8.56%, 8.22%, 7.61%, respectively. 7.50% and 7.11% (P0.05). Conclusion: 1.HMGB1 can promote the proliferation and migration of HPASMC cells, and it has a concentration-dependent effect on HMGB1 and reaches its peak when the concentration of HMGB1 is 100ng/ml. 2. HMGB1 had no effect on the apoptosis of HPASMC. 3. HMGB1 promotes pulmonary vascular remodeling by inducing HPASMC proliferation and migration.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
【共引文献】
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